sop - quy trình vi sinh vật

B Y T

S: 26 /Q-BYT

CNG HA X HI CH NGHA VIT NAM

c lp - T do - Hnh phc

H Ni, ngy 03 thng 01 nm 2013

QUYT NH

V vic ban hnh ti liu

Hng dn quy trnh k thut chuyn ngnh Vi sinh Y hc

B TRNG B Y T

Cn c Lut khm bnh, cha bnh nm 2009;

Cn c Ngh nh s 63/2012/N-CP ngy 31/8/2012 ca Chnh Ph quy

nh chc nng, nhim v, quyn hn v c cu t chc ca B Y t;

Xt Bin bn hp ca Hi ng nghim thu Hng dn Quy trnh k

thut khm bnh, cha bnh chuyn ngnh Vi sinh Y hc ca B Y t;

Theo ngh ca Cc trng Cc Qun l Khm, cha bnh,

QUYT NH:

iu 1. Ban hnh km theo Quyt nh ny ti liu Hng dn quy trnh

k thut chuyn ngnh Vi sinh Y hc, gm 231 quy trnh k thut.

iu 2. Ti liu Hng dn quy trnh k thut chuyn ngnh Vi sinh Y

hc ban hnh km theo Quyt nh ny c p dng ti cc c s khm bnh,

cha bnh.

Cn c vo ti liu hng dn ny v iu kin c th ca n v, Gim

c c s khm bnh, cha bnh xy dng v ban hnh ti liu Hng dn Quy

trnh k thut Vi sinh Y hc ph hp thc hin ti n v.

iu 3. Quyt nh ny c hiu lc k t ngy k ban hnh.

iu 4. Cc ng, b: Chnh Vn phng B, Chnh Thanh tra B, Cc

trng Cc Qun l Khm, cha bnh, Cc trng v V trng cc Cc, V

thuc B Y t, Gim c cc bnh vin, vin c ging bnh trc thuc B Y

t, Gim c S Y t cc tnh, thnh ph trc thuc trung ng, Th trng Y

t cc B, Ngnh v Th trng cc n v c lin quan chu trch nhim thi

hnh Quyt nh ny./.

Ni nhn: - Nh iu 4;

- B trng B Y t ( b/c);

- Cc Th trng BYT;

- Bo him X hi Vit Nam ( phi hp);

- Cng thng tin in t BYT;

- Website Cc KCB;

- Lu VT, KCB.

KT. B TRNG

TH TRNG

k

Nguyn Th Xuyn

B Y T CNG HA X HI CH NGHA VIT NAM

c lp - T do- Hnh Phc

DANH SCH CC QUY TRNH K THUT

CHUYN NGNH VI SINH HC

(Ban hnh km theo Quyt nh s: 26/Q-BYT ngy 03 thng 01 nm 2014

ca B trng B Y t)

TT TN QUY TRNH K THUT

Danh mc xt nghim vi khun

A. Vi khun chung

1 Vi khun nhum soi

2 Vi khun test nhanh

3 Vi khun nui cy v nh danh phng php thng thng

4 Vi khun nui cy v nh danh h thng t ng

5 Vi khun nui cy, nh danh v khng thuc h thng t ng

6 Vi khun khng thuc nh tnh

7 Vi khun khng thuc h thng t ng

8 Vi khun khng thuc nh lng (MIC) (cho 1 loi khng sinh)

9 Vi khun khng sinh phi hp

10 Vi khun k kh nui cy v nh danh

11 Vi khun khng nh

12 Vi khun nh danh PCR

13 Vi khun nh danh gii trnh t gene

14 Vi khun khng thuc PCR

15 Vi khun khng thuc gii trnh t gene

16 Vi h ng rut

B. Mycobacteria

17 AFB trc tip nhum Ziehl-Neelsen

18 AFB trc tip nhum hunh quang

19 Mycobacterium tuberculosis nui cy mi trng lng

20 Mycobacterium tuberculosis nui cy mi trng c

21 Mycobacterium tuberculosis Mantoux

22 Mycobacterium tuberculosis khng thuc hng 1 mi trng c

23 Mycobacterium tuberculosis khng thuc hng 1 mi trng lng

24 Mycobacterium tuberculosis khng thuc hng 2 mi trng c

25 Mycobacterium tuberculosis khng thuc hng 2 mi trng lng

26 Mycobacterium tuberculosis khng thuc PZA mi trng lng

27 Mycobacterium tuberculosis pyrazinamidase

28 Mycobacterium tuberculosis nh danh v khng RMP Xpert

29 Mycobacterium tuberculosis a khng LPA

30 Mycobacterium tuberculosis siu khng LPA

31 Mycobacterium tuberculosis PCR h thng t ng

32 Mycobacterium tuberculosis Real-time PCR

33 Mycobacterium tuberculosis spoligotyping

34 Mycobacterium tuberculosis RFLP typing

35 NTM (Non tuberculosis mycobacteria) nui cy mi trng lng

36 NTM (Non tuberculosis mycobacteria) nui cy mi trng c

37 NTM (Non tuberculosis mycobacteria) nh danh LPA

38 Mycobacterium leprae nhum soi

39 Mycobacterium leprae PCR

40 Mycobacterium leprae mnh sinh thit

C. Vibrio cholerae

41 Vibrio cholerae nhum soi

42 Vibrio cholerae nhum hunh quang

43 Vibrio cholerae nui cy, nh danh v khng thuc

44 Vibrio cholerae PCR

45 Vibrio cholerae gii trnh t gene

D. Neisseria gonorrhoeae

46 Neisseria gonorrhoeae nhum soi

47 Neisseria gonorrhoeae nui cy, nh danh v khng thuc

48 Neisseria gonorrhoeae PCR

49 Neisseria gonorrhoeae Real-time PCR

E. Neisseria meningitidis

50 Neisseria meningitidis nhum soi

51 Neisseria meningitidis nui cy, nh danh v khng thuc

52 Neisseria meningitidis PCR

53 Neisseria meningitidis Real-time PCR

F. Cc vi khun khc

54 Chlamydia test nhanh

55 Chlamydia nhum hunh quang

56 Chlamydia Ab min dch bn t ng

57 Chlamydia PCR

58 Chlamydia Real-time PCR

59 Chlamydia Real-time PCR h thng t ng

60 Clostridium nui cy, nh danh

61 Clostridium difficile min dch bn t ng

62 Clostridium difficile PCR

63 Leptospira test nhanh

64 Mycoplasma pneumoniae Real-time PCR

65 Mycoplasma hominis nui cy, nh danh v khng thuc

66 Rickettsia Ab min dch bn t ng

67 Salmonella Widal

68 Streptococcus pyogenes ASO

69 Treponema pallidum soi ti

70 Treponema pallidum nhum soi

71 Treponema pallidum RPR nh tnh v nh lng

72 Treponema pallidum TPHA nh tnh v nh lng

73 Ureaplasma urealyticum nui cy, nh danh v khng thuc

Danh mc xt nghim virus

A. Virus chung

74 Virus test nhanh

75 Virus Ag min dch bn t ng

76 Virus Ab min dch bn t ng

B. Hepatitis virus

77 HBsAg test nhanh

78 HBsAg min dch bn t ng

79 HBsAg min dch t ng

80 HBsAg khng nh

81 HBsAg nh lng

82 HBsAb min dch bn t ng

83 HBsAb nh lng

84 HBc IgM min dch bn t ng

85 HBc IgM min dch t ng

86 HBc total min dch bn t ng

87 HBc total min dch t ng

88 HBeAg min dch bn t ng

89 HBeAg min dch t ng

90 HBeAb min dch bn t ng

91 HBeAb min dch t ng

92 HBV o ti lng Real-time PCR

93 HBV o ti lng h thng t ng

94 HBV genotype PCR

95 HBV genotype Real-time PCR

96 HBV genotype gii trnh t gene

97 HBV khng thuc Real-time PCR (cho 1 loi thuc)

98 HBV khng thuc gii trnh t gene

99 HCV Ab min dch bn t ng

100 HCV Ab min dch t ng

101 HCV Ag/Ab min dch bn t ng

102 HCV Core Ag min dch t ng

103 HCV PCR

104 HCV o ti lng Real-time PCR

105 HCV o ti lng h thng t ng

106 HCV genotype Real-time PCR

107 HCV genotype gii trnh t gene

108 HAV IgM min dch bn t ng

109 HAV IgM min dch t ng

110 HAV total min dch bn t ng

111 HAV total min dch t ng

112 HDV Ag min dch bn t ng

113 HDV IgM min dch bn t ng

114 HDV Ab min dch bn t ng

115 HEV IgM test nhanh

116 HEV IgM min dch bn t ng

C. HIV

117 HIV Ab test nhanh

118 HIV Ag/Ab test nhanh

119 HIV Ab min dch bn t ng

120 HIV Ag/Ab min dch bn t ng

121 HIV Ag/Ab min dch t ng

122 HIV khng nh

123 HIV o ti lng Real-time PCR

124 HIV o ti lng h thng t ng

125 HIV genotype gii trnh t gene

126 HIV khng thuc gii trnh t gene

D. Dengue virus

127 Dengue virus NS1Ag test nhanh

128 Dengue virus NS1Ag/IgM/IgG test nhanh

129 Dengue virus IgM/IgG test nhanh

130 Dengue virus IgM min dch bn t ng

131 Dengue virus IgG min dch bn t ng

132 Dengue virus PCR

133 Dengue virus serotype PCR

E. Herpesviridae

134 CMV IgM min dch bn t ng

135 CMV IgM min dch t ng

136 CMV IgG min dch bn t ng

137 CMV IgG min dch t ng

138 CMV PCR

139 CMV Real-time PCR

140 CMV o ti lng h thng t ng

141 CMV Avidity

142 HSV 1+2 IgM min dch bn t ng

143 HSV 1+2 IgG min dch bn t ng

144 HSV Real-time PCR

145 VZV Real-time PCR

146 EBV-VCA IgM min dch bn t ng

147 EBV-VCA IgG min dch bn t ng

148 EBV EA-D IgG min dch bn t ng

149 EBV EB-NA IgG min dch bn t ng

150 EBV PCR

151 EBV Real-time PCR

E. Enterovirus

152 EV71 PCR

153 EV71 Real-time PCR

154 EV71 genotype gii trnh t gene

155 Enterovirus PCR

156 Enterovirus genotype gii trnh t gene

F. Cc virus khc

157 Adenovirus Real-time PCR

158 BK/JC virus PCR

159 HPV PCR

160 HPV Real-time PCR

161 HPV genotype Real-time PCR

162 HPV genotype PCR h thng t ng

163 HPV genotype gii trnh t gene

164 Influenza virus A, B test nhanh

165 Influenza virus A, B Real-time PCR

166 Influenza virus A, B gii trnh t gene

167 JEV IgM min dch bn t ng

168 Measles virus Ab min dch bn t ng

169 Rotavirus test nhanh

170 RSV Ab min dch bn t ng

171 RSV Real-time PCR

172 Rubella virus IgM min dch bn t ng

173 Rubella virus IgM min dch t ng

174 Rubella virus IgG min dch bn t ng

175 Rubella virus IgG min dch t ng

176 Rubella virus PCR

177 Rubella virus gii trnh t gene

Danh mc xt nghim KST

A. K sinh trng trong phn

178 Hng cu, bch cu trong phn soi ti

179 Hng cu trong phn test nhanh

180 n bo ng rut soi ti

181 n bo ng rut nhum soi

182 Trng giun, sn soi ti

183 Trng giun soi tp trung

184 Strongyloides stercoralis (giun ln) u trng soi ti

B. K sinh trng trong mu

185 Angiostrogylus cantonensis (Giun trn chut) Ab min dch bn t ng

186 Clonorchis/Opisthorchis (Sn l gan nh) Ab min dch bn t ng

187 Cysticercus cellulosae (Sn ln) Ab min dch bn t ng

188 Entamoeba histolytica (Amip) Ab min dch bn t ng

189 Fasciola (Sn l gan ln) Ab min dch bn t ng

190 Filaria (Giun ch) u trng trong mu nhum soi

191 Gnathostoma Ab min dch bn t ng

192 Plasmodium (K sinh trng st rt) nhum soi nh tnh

193 Plasmodium (K sinh trng st rt) nhum soi nh lng

194 Plasmodium (K sinh trng st rt) Ag test nhanh

195 Strongyloides stercoralis (Giun ln) Ab min dch bn t ng

196 Toxocara (Giun a ch, mo) Ab min dch bn t ng

197 Toxoplasma IgM min dch t ng

198 Toxoplasma IgG min dch t ng

199 Toxoplasma Avidity

C. K sinh trng ngoi da

200 Demodex soi ti

201 Demodex nhum soi

202 Phthirus pubis (Rn mu) soi ti

203 Phthirus pubis (Rn mu) nhum soi

204 Sarcoptes scabies hominis(Gh) soi ti

205 Sarcoptes scabies hominis(Gh) nhum soi

D. K sinh trng trong cc bnh phm khc

206 Cysticercus cellulosae (Sn ln) u trng soi mnh sinh thit

207 Gnathostoma u trng soi mnh sinh thit

208 Pneumocystis jirovecii nhum soi

209 Taenia (Sn dy) soi ti nh danh

210 Toxocara (Giun a ch, mo) soi mnh sinh thit

211 Trichinella spiralis (Giun xon) soi mnh sinh thit

212 Trichomonas vaginalis soi ti

213 Trichomonas vaginalis nhum soi

Danh mc xt nghim Vi nm

214 Vi nm soi ti

215 Vi nm test nhanh

216 Vi nm nhum soi

217 Vi nm nui cy v nh danh phng php thng thng

218 Vi nm nui cy v nh danh h thng t ng

219 Vi nm nui cy, nh danh v khng thuc h thng t ng

220 Vi nm khng nh

221 Vi nm khng thuc nh lng (MIC) (cho 1 loi khng sinh)

222 Vi nm PCR

223 Vi nm gii trnh t gene

Danh mc xt nghim nh gi nhim khun bnh vin

224 Vi sinh vt cy kim tra khng kh

225 Vi sinh vt cy kim tra bn tay

226 Vi sinh vt cy kim tra dng c tit trng

227 Vi sinh vt cy kim tra b mt

228 Vi sinh vt cy kim tra nc sinh hot

229 Vi sinh vt cy kim tra nc thi

230 Vi khun khng thuc - Pht hin ngi mang

231 Vi khun gy nhim trng bnh vin - Pht hin ngun nhim

(Tng s 231 quy trnh k thut) KT. B TRNG

TH TRNG

Nguyn Th Xuyn

MC LC

XT NGHIM VI KHUN

1. Vi khun nhum soi .......................................................................................... 1

2. Vi khun test nhanh ........................................................................................... 4

3. Vi khun nui cy v nh danh phng php thng thng ........................... 7

4. Vi khun nui cy, nh danh h thng t ng ............................................. 10

5. Vi khun nui cy, nh danh v khng thuc h thng t ng ................... 14

6. Vi khun khng thuc nh tnh ...................................................................... 18

7. Vi khun khng thuc h thng t ng ......................................................... 21

8. Vi khun khng thuc nh lng MIC (cho 1 loi khng sinh) .................... 24

9. Vi khun khng sinh phi hp ........................................................................ 28

10. Vi khun k kh nui cy v nh danh ......................................................... 31

11. Vi khun khng nh ..................................................................................... 34

13. Vi khun nh danh gii trnh t gene........................................................... 40

14. Vi khun khng thuc PCR ........................................................................... 44

15. Vi khun khng thuc gii trnh t gene ....................................................... 47

16. Vi h ng rut ........................................................................................... 51

17. AFB trc tip nhum Ziehl-Neelsen ............................................................. 54

18. AFB trc tip nhum hunh quang ............................................................... 58

19. Mycobacterium tuberculosis nui cy mi trng lng ............................... 61

20. Mycobacterium tuberculosis nui cy mi trng c ................................. 66

21. Mycobacterium tuberculosis Mantoux ......................................................... 71

22. Mycobacterium tuberculosis khng thuc hng 1 mi trng c ............... 74

23. Mycobacterium tuberculosis khng thuc hng 1 mi trng lng ............. 79

24. Mycobacterium tuberculosis khng thuc hng 2 mi trng c ............... 83

25. Mycobacterium tuberculosis khng thuc hng 2 mi trng lng ............. 88

26. Mycobacterium tuberculosis khng thuc PZA mi trng lng ................ 92

27. Mycobacterium tuberculosis pyrazinamidase ............................................... 96

28. Mycobacterium tuberculosis nh danh v khng RMP XPERT ................. 99

29. Mycobacterium tuberculosis a khng LPA ............................................... 102

30. Mycobacterium tuberculosis siu khng LPA ............................................ 107

31. Mycobacterium tuberculosis PCR h thng t ng .................................. 113

32. Mycobacterium tuberculosis Real-time PCR ............................................. 116

33. Mycobacterium tuberculosis Spoligotyping ............................................... 120

34. Mycobacterium tuberculosis RFLP typing ................................................. 125

35. NTM (Non Tuberculosis Mycobacteria) nui cy mi trng lng .......... 131

36. NTM (Non Tuberculosis Mycobacteria) nui cy mi trng c ............ 136

37. NTM (Non Tuberculosis Mycobacteria) LPA ............................................ 141

38. Mycobacterium leprae nhum soi .............................................................. 145

39. Mycobacterium leprae PCR ....................................................................... 148

40. Mycobacterium leprae mnh sinh thit ...................................................... 152

41. Vibrio cholerae nhum soi ......................................................................... 154

42. Vibrio cholerae nhum hunh quang ......................................................... 157

43. Vibrio cholerae nui cy, nh danh v khng thuc ................................. 160

44. Vibrio cholerae PCR ................................................................................... 163

45. Vibrio cholerae gii trnh t gene ............................................................... 166

46. Neisseria gonorrhoeae nhum soi .............................................................. 169

47. Neisseria gonorrhoeae nui cy, nh danh v khng thuc ..................... 172

48. Neisseria gonorrhoeae PCR ....................................................................... 175

49. Neisseria gonorrhoeae Real-time PCR ...................................................... 178

50. Neisseria meningitidis nhum soi ............................................................... 182

51. Neisseria meningitidis nui cy, nh danh v khng thuc ...................... 185

52. Neisseria menigititdis PCR ......................................................................... 188

54. Chlamydia test nhanh .................................................................................. 195

55. Chlamydia nhum hunh quang ................................................................. 198

56. Chlamydia Ab min dch bn t ng ........................................................ 201

57. Chlamydia PCR .......................................................................................... 205

58. Chlamydia Real-time PCR.......................................................................... 209

59. Chlamydia Real-time PCR h thng t ng ............................................. 213

60. Clostridium nui cy, nh danh ................................................................. 216

61. Clostridium difficile min dch bn t ng ............................................... 219

62. Clostridium difficile PCR............................................................................ 222

63. Leptospira test nhanh .................................................................................. 225

64. Mycoplasma pneumoniae Real-time PCR .................................................. 227

65. Mycoplasma hominis nui cy, nh danh v khng thuc ........................... 231

66. Ricketsia Ab min dch bn t ng ........................................................... 234

67. Salmonella Widal ........................................................................................ 237

68. Streptococcus pyogenes ASO ..................................................................... 240

69. Treponema pallidum soi ti ...................................................................... 243

70. Treponema pallidum nhum soi ................................................................. 245

71. Treponema pallidum RPR nh tnh v nh lng .................................... 248

72. Treponema pallidum TPHA nh tnh v nh lng ................................. 251

73. Ureaplasma urealyticum nui cy, nh danh v khng thuc ...................... 255

XT NGHIM VIRUS

74. Virus test nhanh ........................................................................................... 258

75. Virus Ag min dch t ng ........................................................................ 260

76. Virus Ab min dch t ng ........................................................................ 264

77. HBsAg test nhanh ....................................................................................... 268

78. HBsAg min dch bn t ng .................................................................... 271

79. HBsAg min dch t ng........................................................................... 275

80. HBsAg khng nh ...................................................................................... 279

81. HBsAg nh lng ...................................................................................... 283

82. HBsAb min dch bn t ng .................................................................... 287

83. HBsAb nh lng ...................................................................................... 291

84. HBc IgM min dch bn t ng ................................................................ 295

85. HBc IgM min dch t ng ....................................................................... 299

86. HBc total min dch bn t ng ................................................................ 303

87. HBc total min dch t ng ....................................................................... 307

88. HBeAg min dch bn t ng ................................................................... 311

89. HBeAg min dch t ng .......................................................................... 315

90. HBeAb min dch bn t ng ................................................................... 318

91. HBeAb min dch t ng .......................................................................... 322

92. HBV o ti lng Real-time PCR .............................................................. 325

93. HBV o ti lng h thng t ng ............................................................ 329

94. HBV genotype PCR .................................................................................... 333

95. HBV genotype Real-time PCR ................................................................... 336

96. HBV genotype gii trnh t gene ................................................................ 340

97. HBV khng thuc Real-time PCR (Cho 1 loi thuc) ................................ 343

98. HBV khng thuc gii trnh t gene ........................................................... 346

99. HCV Ab min dch bn t ng ................................................................. 349

100. HCV Ab min dch t ng ...................................................................... 353

101. HCV Ag/Ab min dch bn t ng ......................................................... 357

102. HCV Core Ag min dch t ng ............................................................. 361

103. HCV PCR .................................................................................................. 365

104. HCV o ti lng Real-time PCR ............................................................ 368

105. HCV o ti lng h thng t ng ......................................................... 372

106. HCV genotype Real-time PCR ................................................................. 375

107. HCV genotype gii trnh t gene .............................................................. 379

108. HAV IgM min dch bn t ng ............................................................. 382

109. HAV IgM min dch t ng .................................................................... 386

110. HAV total min dch bn t ng ............................................................. 390

111. HAV total min dch t ng ................................................................... 394

112. HDV Ag min dch bn t ng ............................................................... 398

113. HDV IgM min dch bn t ng ............................................................. 402

114. HDV Ab min dch bn t ng ............................................................... 406

115. HEV IgM test nhanh ................................................................................. 409

116. HEV IgM min dch bn t ng ............................................................. 412

117. HIV Ab test nhanh .................................................................................... 415

118. HIV Ag/Ab test nhanh .............................................................................. 418

120. HIV Ag/Ab min dch bn t ng .......................................................... 425

121. HIV Ag/Ab min dch t ng ................................................................. 429

122. HIV khng nh ........................................................................................ 433

123. HIV o ti lng Real-time PCR.............................................................. 437

124. HIV o ti lng h thng t ng ........................................................... 440

125. HIV genotype gii trnh t gene ............................................................... 444

128. Dengue virus NS1Ag/IgM/IgG test nhanh ............................................... 453

129. Dengue virus IgM/IgG test nhanh ............................................................ 456

130. Dengue virus IgM min dch bn t ng ................................................ 459

131. Dengue virus IgG min dch bn t ng ................................................. 463

132. Dengue virus PCR .................................................................................... 467

133. Dengue virus serotype PCR ...................................................................... 470

134. CMV IgM min dch bn t ng ............................................................. 473

135. CMV IgM min dch t ng ................................................................... 477

136. CMV IgG min dch bn t ng ............................................................. 481

137. CMV IgG min dch t ng .................................................................... 485

138. CMV PCR ................................................................................................. 489

139. CMV Real-time PCR ................................................................................ 492

140. CMV o ti lng h thng t ng ......................................................... 495

141. CMV Avidity ............................................................................................ 498

142. HSV 1 + 2 IgM min dch bn t ng .................................................... 502

143. HSV 1+ 2 IgG min dch bn t ng ...................................................... 506

144. HSV Real-time PCR ................................................................................. 510

145. VZV Real-time PCR ................................................................................. 514

146. EBV-VCA IgM min dch bn t ng .................................................... 518

147. EBV-VCA IgG min dch bn t ng ..................................................... 522

148. EBV EA-D IgG min dch bn t ng .................................................... 526

149. EBV EB-NA IgG min dch bn t ng ................................................. 530

150. EBV PCR .................................................................................................. 534

151. EBV Real-time PCR ................................................................................. 537

152. EV71 PCR ................................................................................................. 541

155. Enterovirus PCR........................................................................................ 551

156. Entrovirus genotype gii trnh t gene ...................................................... 554

157. Adenovirus Real-time PCR ....................................................................... 557

158. BK/JC virus PCR ...................................................................................... 561

159. HPV PCR .................................................................................................. 565

160. HPV Real-time PCR ................................................................................. 568

161. HPV Genotype Real-time PCR ................................................................. 572

162. HPV genotype PCR h thng t ng ...................................................... 575

163. HPV genotype gii trnh t gene ............................................................... 579

164. Influenza virus A, B test nhanh ................................................................. 582

165. Influenza virus A, B Real-time PCR ......................................................... 584

166. Influenza virus A, B gii trnh t gene ..................................................... 587

167. JEV IgM min dch bn t ng ............................................................... 590

168. Measle virus Ab min dch bn t ng ................................................... 593

169. Rotavirus test nhanh .................................................................................. 596

170. RSV Ab min dch bn t ng ................................................................ 599

171. RSV Real-time PCR ................................................................................. 602

172. Rubella virus IgM min dch bn t ng ................................................ 605

173. Rubella virus IgM min dch t ng ....................................................... 609

174. Rubella virus IgG min dch bn t ng ................................................. 613

175. Rubella virus IgG min dch t ng ....................................................... 617

176. Rubella virus PCR .................................................................................... 621

177. Rubella virus gii trnh t gene................................................................. 624

XT NGHIM K SINH TRNG - VI NM

178. Hng cu, bch cu trong phn soi ti ................................................... 627

179. Hng cu trong phn test nhanh ............................................................... 630

180. n bo ng rut soi ti ..................................................................... 633

181. n bo ng rut nhum soi ................................................................ 636

182. Trng giun, sn soi ti ............................................................................ 639

183. Trng giun soi tp trung ........................................................................... 642

184. Strongyloides stercoralis (Giun ln) u trng soi ti ........................... 645

185. Angiostrongylus cantonensis (Giun trn chut) Ab min dch bn t

ng .......................................................................................................... 648

186. Clonorchis/Opisthorchis (Sn l gan nh) Ab min dch bn t

ng .......................................................................................................... 651

187. Cysticercus cellulosae (Sn ln) Ab min dch bn t ng.................... 654

188. Entamoeba histolytica (Amip) Ab min dch bn t ng ....................... 657

189. Fasciola (Sn l gan ln) Ab min dch bn t ng ............................... 660

190. Filaria (Giun ch) u trng trong mu nhum soi .................................... 663

191. Gnathostoma(Giun u gai) Ab Min dch bn t ng.......................... 666

192. Plasmodium (K sinh trng st rt) nhum soi nh tnh ........................ 669

193. Plasmodium (K sinh trng st rt) nhum soi nh lng ..................... 672

194. Plasmodium (K sinh trng st rt) Ag test nhanh .................................. 675

195. Strongyloides stercoralis (Giun ln) Ab min dch bn t ng ........... 677

196. Toxocara (Giun a ch mo) Ab min dch bn t ng ....................... 680

197. Toxoplasma IgM min dch t ng ......................................................... 683

198. Toxoplasma IgG min dch t ng ......................................................... 687

199. Toxoplasma Avidity min dch t ng ................................................... 691

200. Demodex soi ti ...................................................................................... 695

201. Demodex nhum soi .................................................................................. 698

202. Phthirus pubis (Rn mu) soi ti .............................................................. 701

203. Phthirus pubis (Rn mu) nhum soi ......................................................... 703

204. Sarcoptes scabies hominis (Gh) soi ti ................................................. 706

205. Sarcoptes scabies hominis (Gh) nhum soi ............................................ 709

206. Cysticercus cellulosae (Sn ln) u trng soi mnh sinh thit ................. 712

207. Gnathostoma u trng soi mnh sinh thit ............................................... 715

208. Taenia (Sn dy) soi ti nh danh ........................................................ 718

209. Toxocara (Giun a ch, mo) soi mnh sinh thit ................................ 721

210. Trichinella spiralis (Giun xon) soi mnh sinh thit ................................ 724

211. Trichomonas vaginalis soi toi ................................................................ 727

212. Trichomonas vaginalis nhum soi ............................................................ 730

213. Pneumocystis jirovecii nhum soi............................................................. 733

214. Vi nm soi ti .......................................................................................... 736

215. Vi nm test nhanh...................................................................................... 739

216. Vi nm nhum soi ..................................................................................... 742

217. Vi nm nui cy v nh danh bng phng php thng thng ............. 745

218. Vi nm nui cy v nh danh h thng t ng ...................................... 748

219. Vi nm nui cy, nh danh v khng thuc h thng t ng ................ 751

220. Vi nm khng nh (tham chiu)............................................................... 754

221. Vi nm khng thuc nh lng (MIC) (Cho 1 loi khng sinh) ............. 757

222. Vi nm PCR .............................................................................................. 760

223. Vi nm gii trnh t gene .......................................................................... 763

XT NGHIM NH GI NHIM KHUN BNH VIN

224. Vi sinh vt cy kim tra khng kh ........................................................... 766

225. Vi sinh vt cy kim tra bn tay ................................................................ 770

226. Vi sinh vt cy kim tra dng c tit trng .......................................... 773

227. Vi sinh vt cy kim tra b mt................................................................. 776

228. Vi sinh vt cy kim tra nc sinh hot .................................................... 779

229. Vi sinh vt cy kim tra nc thi ............................................................ 783

230. Vi khun khng thuc - Pht hin ngi mang ........................................ 787

231. Vi khun gy nhim trng bnh vin - Pht hin ngun nhim ............... 841

CC T VIT TT

BCN: Bch cu a nhn

CLIA: Chemiluminescence immunoassay

CLSI: Clinical and laboratory standards institute

CO: Cut off

DNA: Deoxyribonucleic acid

ELISA: Enzyme-linked immunosorbant assay

EQAS: External quality assessment scheme

HBV: Hepatitis B virus

HCV: Hepatitis C virus

HIV: Human immunodeficiency virus

HPC: High positive control

HT: Huyt thanh

I: Intermediate

IU: International unit

KKT: Khng khng th

KN-KT: Khng nguyn - Khng th

KST: K sinh trng

LPC: Low positive control

NC: Negative control

OD: Optical density

PC: Positive control

PCR: Polymerase chain reaction

QC: Quality control

R: Resistant

RNA: Ribonucleic acid

RT PCR: Reverse transcription polymerase chain reaction

S: Susceptible

VD: V d

VK: Vi khun

VSV: Vi sinh vt

1

1. Vi khun nhum soi

I. MC CH V NGUYN L

1. Mc ch

Nhn nh s b hnh nh vi khun v cc hnh nh t bo (nu c) trc

tip t bnh phm.

2. Nguyn l

nh gi hnh th, kch thc, tnh cht bt mu, cch sp xp ca vi

khun v cc hnh nh t bo (nu c) bng k thut nhum v soi di knh

hin vi quang hc.

II. CHUN B

1. Ngi thc hin

- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.

- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.

2. Phng tin, ha cht

Phng tin, ha cht nh v d di y hoc tng ng.

2.1. Trang thit b

- My ly tm (cn cho mt s loi bnh phm)

- T an ton sinh hc cp 2

- Knh hin vi quang hc

- Dng c sy lam (nu c)

2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)

STT Chi ph ha cht, vt t tiu hao n v S lng

1 L ly bnh phm L 1,000

2 Que cy Ci 1,000

3 Lam knh Ci 2,000

4 Du soi knh ml 1,000

5 Cn 96 lau knh ml 1,000

6 Nc mui sinh l ml 5,000

7 Thuc nhum fuchsin ml 5,000

8 Thuc nhum tm gentian ml 5,000

9 Cn ty 95% ml 10,000

2

10 Lugol ml 5,000

11 Thuc nhum xanh methylen ml 5,000

12 Bng Kg 0,001

13 Cn 90 (v sinh dng c) ml 10,000

14 n cn Ci 0,0001

15 Panh Ci 0,0001

16 Khay ng bnh phm Ci 0,0001

17 Hp vn chuyn bnh phm Ci 0,0001

18 M Ci 0,020

19 Khu trang Ci 0,020

20 Gng tay i 3,000

21 Gng tay x l dng c i 0,020

22 Qun o bo h B 0,001

23 Acid ngm lam ml 10,000

24 ng nghim thy tinh ng 1,000

25 Bt vit knh Ci 0,020

26 Bt bi Ci 0,010

27 Bt la Ci 0,010

28 S lu kt qu xt nghim T 0,001

29 Cn st trng tay nhanh ml 1,000

30 Dung dch nc ra tay ml 8,000

31 Khn lau tay Ci 0,030

32 Giy tr kt qu xt nghim T 2,000

33 QC (nu thc hin) * 0,1

34 EQAS (nu thc hin) * 0,005

* Ghi ch:

- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni

kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng

10 mu cho 1 ln tin hnh k thut).

- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh

ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi

s ln ngoi kim trung bnh 2 ln/1 nm).

3. Bnh phm

3

Cc loi bnh phm c ch nh xt nghim vi khun.

4. Phiu xt nghim

in y thng tin theo mu phiu yu cu.

III. CC BC TIN HNH

Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.

- Nh dung dch tm gentian, ph kn ni dn phin, duy tr 1 - 2 pht

- dung dch tm gentian, ra tiu bn di vi nc chy nh

- Nh dung dch lugol, 30 giy

- dung dch lugol, ra nc

- Ty mu: nh vi git cn 95% ln tiu bn, nghing i nghing li cho

cn chy t cnh n sang cnh kia. Khi thy mu tm trn lam knh va phai

ht th ra nc ngay.

- Nh dung dch fuchsin, 1 - 2 pht

- Ra nc k, kh tiu bn, soi knh hin vi.

IV. NHN NH KT QU

Ch c kt qu khi QC t tiu chun.

1. nh gi hnh nh vi khun trn tiu bn nhum Gram

- Soi di vt knh du (x100)

- Din gii v c kt qu:

Vi khun Gram (+) bt mu tm sm ca gentian.

Vi khun Gram (-) bt mu ca fuchsin.

2. nh gi hnh nh t bo trn tiu bn nhum n (nu c).

V. NHNG SAI ST V X TR

Gram dng gi: Ty cn cha thi gian

Gram m gi:

+ Tui ca mu cy vi khun nh hng ln tnh cht nhum Gram cc mu cy vi khun thi gian qu lu.

+ Ty cn qu lu v trng khng k.

Nhum li tiu bn khi nghi ng kt qu khng chnh xc.

4

2. Vi khun test nhanh

I. MC CH V NGUYN L

1. Mc ch

Xc nh nhanh s c mt ca vi khun gy bnh trong bnh phm thng

qua pht hin s c mt ca khng nguyn vi khun.

2. Nguyn l

Pht hin khng nguyn vi khun da trn phn ng kt hp c hiu

gia khng nguyn vi khng huyt thanh mu.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1 . Trang thit b



1 Bng Kg 0,001

2 Dy gar Ci 0,001

3 Cn ml 1,000

4 Bm kim tim Ci 1,000

5 Panh Ci 0,0001



8 Tube ng bnh phm Ci 1,000

9 Sinh phm chn on Test 1,000

10 u cn 200 ul Ci 2,000

11 Giy thm Cun 0,100

12 Giy xt nghim T 2,000


5


15 Bt bi Ci 0,010

16 M Ci 0,020


18 Gng tay i 0,100


20 Qun o bo h B 0,005



23 Dung dch kh trng ml 10,000




Thc hin xt nghim 05 mu bnh phm/ln.

* Ghi ch:







3. Bnh phm

Cc loi bnh phm ch nh lm test nhanh.

4. Phiu xt nghim

in y thng tin theo mu phiu yu cu

III. CC BC THC HIN


1. Ly bnh phm

Theo ng quy nh ca chuyn ngnh Vi sinh (Xem Ph lc 1).

2. Tin hnh k thut

B sinh phm Slidex Meningite Kit 5 (VD) pht hin 5 cn nguyn vi

khun gy vim mng no.

6

Cc bc

thc hin

K thut

1. a sinh phm ra ngoi nhit phng

nh s th t cc bnh phm v s th t trn tm knh

2. Lc nh nhng l cha ht latex, khng c lc qu mnh

3. Dng pipet nh 1 git dch no ty ca ngi bnh vo tng ng nh s trn phin knh. Nh chng dng v

chng m

4. Lc nh nhng l c cha ht latex v nh mt git vo cnh git dch no ty ca ngi bnh

5. Trn u 2 loi vi nhau bng que trn ph u b mt ca mi .

6. Lc u c phin knh bng tay hoc dng my lc 80-100 vng trong 10 pht

7. c kt qu

IV. NHN NH KT QU

- chng dng: c nhin tng ngng kt ht latex

- chng m: khng c nhin tng ngng kt, hn dch nhn thy mn, ng nht

- khng c hin tng ngng kt, hn dch nhn thy mn, ng nht: m tnh

- c hin tng ngng kt, hn dch nhn thy th, c ht ngng kt r trn nn en.


Lng bnh phm a vo qu nhiu hoc qu t u c th nh hng

n kt qu.

Do vy, phi tin hnh ly th tch bnh phm ng theo yu cu (Xem

Ph lc 6).

7

3. Vi khun nui cy v nh danh

phng php thng thng

I. MC CH V NGUYN L

1. Mc ch

Pht hin v nh danh vi khun gy bnh bng phng php nui cy

kinh in.

2. Nguyn l

Vi khun c nh danh da vo c im nui cy, mt s tnh cht

chuyn ha, cc c im v hnh thi hc v c th kt hp vi tnh cht khng

nguyn.

II. CHUN B

1. Ngi thc hin



3. Phng tin, ha cht


2.1 .Trang thit b



- T m thng

- T m CO2

- My tnh ci phn mm c API (nu c)

2.2 .Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)



2 Que cy Ci 1,000

3 Lam knh Ci 2,000




8



9 Cn ty 95 ml 10,000

10 Lugol ml 5,000


12 Mi trng nui cy a 1,500

13 B sinh vt ha hc (b API ) Test 0,500

14 Bng Kg 0,001

15 Cn 90 (v sinh dng c) ml 10,000

16 n cn Ci 0,0001

17 Panh Ci 0,0001



20 M Ci 0,020


22 Gng tay i 3,000


24 Qun o bo h B 0,001




28 Bt bi Ci 0,010

29 Bt la Ci 0,010








Mi trng nui cy v ha cht nh danh vi khun c tnh trn t l

dng tnh trung bnh l 50 % cho cc loi bnh phm.

9

* Ghi ch:







3. Bnh phm

Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh.

4. Phiu xt nghim


III. CC BC TIN HNH


1. Ly bnh phm


2. Tin hnh k thut

- Nhum soi bnh phm, nh gi s b

- Nui cy bnh phm vo mi trng phn lp

- m qua m

- Bt khun lc nghi ng

- Nhum soi, th nghim cc th nghim sinh vt ha hc n gin v nh danh bng cc b sinh vt ha hc (b API )

IV. NHN NH KT QU

- Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi.

- m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh.


Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy,

khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c

ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn

nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny.

Nu c yu cu tm cn nguyn vi khun him gp, phi ghi yu cu c

th.

Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a

n kt qu m tnh hoc dng tnh gi (Xem Ph lc 6).

10

4. Vi khun nui cy, nh danh h thng t ng

I. MC CH V NGUYN L

1. Mc ch

Pht hin vi khun gy bnh bng phng php nui cy, nh danh trn

h thng t ng.

2. Nguyn l

Vi khun c nh danh bng my t ng da trn mt s tnh cht

chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu.

II. CHUN B

1. Ngi thc hin

- Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.


2. Phng tin, ha cht


2.1. Trang thit b


- T m thng

- T m CO2

- H thng nh danh t ng

- My o c

- L hp t





2 Que cy Ci 2,000

3 Lam knh Ci 2,000




11



9 Cn ty 95 ml 10,000

10 Lugol ml 5,000



13 Panel nh danh theo h thng my t ng Test 0,500

14 B Panel chng B 0,0002


16 Bng Kg 0,001

17 Cn 90 (v sinh dng c) ml 10,000

18 n cn Ci 0,0001

19 Panh Ci 0,0001



22 M Ci 0,020


24 Gng tay i 3,000


26 Qun o bo h B 0,001




30 Bt bi Ci 0,010

31 Bt la Ci 0,010








12

Mi trng nui cy c tnh trn t l dng l 50 % so vi tng s

bnh phm gi xt nghim.

B Panel s dng kim chun c tnh trn 5000 Test/1 nm.

* Ghi ch:







3. Bnh phm


4. Phiu xt nghim


II. CC BC TIN HNH


1. Ly bnh phm

Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 1).

2. Tin hnh k thut



- m qua m


- Nhum soi, th nghim cc tnh cht sinh vt ha hc n gin v nh danh bng h thng t ng.

IV. NHN NH KT QU








Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th.

13

Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a n

kt qu m tnh hoc dng tnh gi (Xem Ph lc 6).

14

5. Vi khun nui cy, nh danh v khng thuc h thng

t ng

I. MC CH V NGUYN L

1. Mc ch

Pht hin vi khun gy bnh bng phng php nui cy, nh danh v

khng thuc trn h thng t ng.

2. Nguyn l


chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu; v

xc nh mc nhy cm vi khng sinh bng phng php khng sinh pha

long.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1. Trang thit b


- T m thng

- T m CO2


- My o c

- L hp t





2 Lam knh Ci 2,000

3 Que cy Ci 2,000


15



7 Thuc nhum Fuchsin ml 5,000

8 Thuc nhum tm Gentian ml 5,000

9 Cn ty 95 ml 10,000

10 Lugol ml 5,000






16 Bng Kg 0,001

17 Cn 90 (v sinh dng c) ml 10,000

18 n cn Ci 0,0001

19 Panh Ci 0,0001



22 M Ci 0,020


24 Gng tay i 3,000


26 Qun o bo h B 0,001




30 Bt bi Ci 0,010

31 Bt la Ci 0,010






16



Mi trng nui cy c tnh trn t l dng l 50 % so vi tng s

bnh phm gi xt nghim.


* Ghi ch:







3. Bnh phm


4. Phiu xt nghim


III. CC BC TIN HNH


1. Ly bnh phm


2. Tin hnh k thut



- m qua m



IV. NHN NH KT QU






17



S dng sai panel nh danh cho loi vi khun cn xc nh. Cn chn la

chnh xc panel nh danh cho tng loi vi khun.




18

6. Vi khun khng thuc nh tnh

I. MC CH V NGUYN L

1. Mc ch

Xc nh mc nhy cm vi khng sinh ca cc chng vi khun gy

bnh.

2. Nguyn l

Mc nhy cm vi khng sinh ca cc chng vi khun th nghim

c nh gi bng phng php khng sinh khuch tn trong thch. S pht

trin ca vi khun s b c ch khi khng sinh t n mt nng nht nh.

ng knh vng c ch t l thun vi mc nhy cm c phin gii ra

cc phn loi S (sensitive nhy cm), I (intermediate trung gian), hoc R

(resistant khng) khi so snh vi bng chun CSLI cp nht hng nm.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1 . Trang thit b

- T m


- My lc

- ng c chun McFarland 0.5



1 Khoanh giy khng sinh Khoanh 22,000

2 Que cy Ci 1,000

3 Thch Muller Hinton a 3,000

4 Thch mu a 1,000

5 Giy khng sinh T 2,000

6 Bng Kg 0,001

7 Cn 90 (v sinh dng c) ml 10,000

19

8 Panh Ci 0,0001



11 M Ci 0,050


13 Gng tay i 3,000


15 Qun o bo h B 0,001



18 Tm bng v trng Ci 1,000


20 Bt bi Ci 0,010

21 Bt la Ci 0,010

22 S lu kt qu xt nghim Quyn 0,001






* Ghi ch:







3. Bnh phm

Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi.

4. Phiu xt nghim


III. CC BC TIN HNH


20

1. Ly bnh phm

Chng vi khun cn th nghim c nui cy thun nht trong iu

kin ti u v ang giai on pht trin mnh (nui cy sau 16-24 gi).

2. Tin hnh k thut

2.1. Pha huyn dch vi khun

Dng que cy ly vi khun t 3-5 khun lc c hnh thi ging nhau nghin

u vo ng nc mui sinh l 5 ml, lc u trn my lc c huyn dch

ng nht. So snh c ca huyn dch vi khun vi c ca ng

McFarland 0,5.

2.2 . Dn u canh khun ln mt a thch

Dng tm bng v trng nhng vo ng huyn dch vi khun pha trn,

p nh v xoay trn tm bng trn thnh bn ca ng huyn dch vi khun

loi bt phn huyn dch vi khun thm vo u tm bng. Sau , ria u

que tm bng trn ton b mt a thch Mueller-Hinton sao cho vi khun c

dn u ln trn ton b b mt a thch

2.3. t cc khoanh giy khng sinh ln mt thch

Cc khoanh giy sau khi t cn c n xung va phi m bo chng

c tip xc hon ton vi mt thch. Trong vng 15 pht sau khi t khoanh

giy khng sinh, cc a thch phi c lt p trong t m 35C.

2.4. a thch trong t m 16 - 24 gi

IV. NHN NH KT QU

Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t.

o ng knh vng c ch (bao gm c ng knh ca khoanh giy

khng sinh) tnh theo mm.

Phin gii ng knh vng c ch ra kt qu S, I, R theo hng dn ca

CLSI cp nht hng nm.


Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d

nui cy, khng p dng cho cc vi khun k kh bt buc.

Sai st c th gp khi:

- Ln hai hay nhiu chng vi khun

- Vi khun mc qu dy hoc qu tha.

Phi tin hnh lm li khi thy cc hin tng nh trn.

21

7. Vi khun khng thuc h thng t ng

I. MC CH V NGUYN L

1. Mc ch

Xc nh mc nhy cm vi khng sinh ca cc chng vi khun gy

bnh.

2. Nguyn l

Mc nhy cm vi khng sinh ca cc chng vi khun th nghim

c nh gi da trn gi tr ca nng c ch ti thiu - MIC. Gi tr MIC

s c phin gii ra phn loi S (sensitive nhy cm), I (intermediate trung

gian), hoc R (resistant khng) nh h thng t ng.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1 . Trang thit b

- My nh danh, khng sinh t ng (v d H thng Vitek 2 Compact)


- My vi tnh

- My in

2.2 . Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)


1 Th khng sinh Th 1,000

2 Que cy Ci 1,000

3 Blood agar base 500G ml 75,000

4 Dng c ly bnh phm L 1,000

5 Mac Conkey agar ml 75,000


7 M Ci 0,030


22

9 Gng tay i 3,000


11 Qun o bo h Ci 0.0005





16 Presept Vin 1,000



* Ghi ch:







3. Bnh phm


4. Phiu xt nghim


III. CC BC TIN HNH


1. Ly bnh phm

Chng vi khun cn th nghim c nui cy thun nht trong iu

kin ti u v ang giai on pht trin mnh (nui cy sau 16-24 gi).

2. Tin hnh k thut

- Chun b mu

- Np ng nghim v card test ng loi vo cassette

- Nhp mu vo my

- Khai bo thng tin

- To thng tin bnh nhn

- Xem kt qu ca mu

23

IV. NHN NH KT QU


Ghi nhn cc kt qu S, I hay R cng vi gi tr MIC.




Sai st c th gp khi:


- Chn th lm khng sinh khng ph hp vi chng vi khun cn th

nghim

- Chng vi khun sau khi pha khng c a vo my trong vng 30 pht


24

8. Vi khun khng thuc nh lng MIC

(cho 1 loi khng sinh)

I. MC CH V NGUYN L

1. Mc ch

Xc nh mc nhy cm v nng c ch ti thiu ca khng sinh

vi chng vi khun gy bnh.

2. Nguyn l

Etest l thanh plastic kch thc 5 x 57 mm c phn b di MIC theo

g/mL v c m code tn khng sinh. C 15 nng khng sinh c pha

long bc 2 gn c nh mt mt ca bng plastic. Khi bng Etest c t ln

mt thch dn vi khun, khng sinh c bc nng nhanh chng khuch

tn trong thch. Sau khi nui cy qua m, s xut hin vng c ch hnh elip

i xng qua thanh plastic. Gi tr MIC c xc nh trc tip ti im ct ca

hnh elip vi thanh Etest. Gi tr MIC s c phin gii ra phn loi S

(sensitive nhy cm), I (intermediate trung gian), hoc R (resistant

khng) khi so snh vi bng chun CLSI cp nht hng nm.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1 . Trang thit b

- T m


- My lc


2.2 . Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)


1 Thanh giy khng sinh Ci 1,000

2 Que cy Ci 1,000


4 Thch mu a 1,000

25


6 Bng Kg 0,001


8 Panh Ci 0,0001



11 M Ci 0,050


13 Gng tay i 3,000


15 Qun o bo h B 0,001





20 Bt bi Ci 0,010

21 Bt la Ci 0,010







* Ghi ch:







3 Bnh phm


26

4 Phiu xt nghim


III. CC BC TIN HNH


1. Ly bnh phm

Chng vi khun cn th nghim c nui cy thun nht trong iu kin ti

u v ang giai on pht trin mnh (nui cy sau 16-24 gi).

2. Tin hnh k thut

2.1. Pha huyn dch vi khun

Dng que cy ly vi khun t 3-5 khun lc c hnh thi ging nhau nghin

u vo ng nc mui sinh l 5 ml, lc u trn my lc c huyn dch

ng nht. So snh c ca huyn dch vi khun vi c ca ng

McFarland 0,5.

2.2. Dn u canh khun ln mt a thch

Dng tm bng v trng nhng vo ng huyn dch vi khun pha trn,

p nh v xoay trn tm bng trn thnh bn ca ng huyn dch vi khun

loi bt phn huyn dch vi khun thm vo u tm bng. Sau , ria u

que tm bng trn ton b mt a thch Mueller-Hinton sao cho vi khun c

dn u ln trn ton b b mt a thch

2.3. t cc khoanh giy khng sinh ln mt thch

t thanh Etest ln mt thch sao cho mt c ghi di nng hng ln trn

v phi m bo ton b b mt ca thanh Etest c tip xc hon ton vi

mt thch. Khi t xong thanh Etest khng c dch chuyn thanh Etest

khi v tr.

2.4. a thch trong t m 16 - 24 gi

IV. NHN NH KT QU

1. Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t.

2. c kt qu

Sau m 16 24 h v khi thy r vi khun mc, c gi tr MIC im ct

ca hnh elip vi thanh E-test. Lm trn gi tr MIC im gia hai bc pha

long ln gi tr cao hn mt bc trc khi phin gii kt qu.

Phin gii kt qu MIC ra gi tr S, I hoc R theo ti liu ca CLSI cp

nht hng nm.

Vi cc khng sinh dit khun, phi c gi tr MIC ti im vi khun b

c ch hon ton. Vi cc khng sinh km khun, phi c gi tr MIC ti im

vi khun b c ch 80%.


27



Sai st c th gp khi:




28

9. Vi khun khng sinh phi hp

I. MC CH V NGUYN L

1. Mc ch

Tm cp khng sinh phi hp c tc dng hip ng trn chng vi khun

gy bnh.

2. Nguyn l

Xc nh tc dng hip ng, i khng hoc khng khc bit (gi tr

MIC) khi th nghim phi hp 2 khng sinh khc nhau vi vi khun gy bnh

bng thanh E-test da trn ch s FIC (Fractional Inhibitory Concentration).

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1. Trang thit b

- T m


- My lc




1 Thanh giy khng sinh Ci 4,000


3 Que cy Ci 1,000

4 Thch mu a 2,000


6 Bng Kg 0,001


8 Panh Ci 0,0001


29


11 M Ci 0,050


13 Gng tay i 3,000


15 Qun o bo h bo h B 0,001





20 Bt bi Ci 0,010

21 Bt la Ci 0,010







* Ghi ch:







3. Bnh phm


4. Phiu xt nghim


III. CC BC TIN HNH


- Pha huyn dch vi khun c c tng ng vi c McFarland 0.5

30

- Dn u canh khun ln mt 2 a thch Mueller-Hinton v cy thun trn 1 a thch mu

- t 2 thanh E-test ln 2 a thch dn u vi khun

- cc a thch trong t m 16 - 18 gi

- c kt qu gi tr MIC ca c hai khng sinh

- Pha huyn dch vi khun c c tng ng vi c McFarland 0.5 ri dn u canh khun ln mt 1 a thch Mueller-Hinton v cy thun trn 1

a thch mu

- t 2 thanh E-test vung gc vi nhau, giao nhau ti gi tr MIC ca mi thanh c bc 5 ln a thch dn u vi khun

- cc a thch trong t m 16 - 18 gi

- c kt qu gi tr MIC ca c hai khng sinh

IV. NHN NH KT QU


c kt qu:

- Tnh ch s FIC (Fractional Inhibitory Concentration)

FIC ca khng sinh A = MIC ca khng sinh A trong th nghim phi

hp/MIC ca khng sinh A th nghim ring

FIC ca khng sinh B = MIC ca khng sinh B trong th nghim phi

hp/MIC ca khng sinh B th nghim ring

Ch s FIC = FIC ca khng sinh A/ FIC ca khng sinh B

- Phin gii kt qu ch s FIC

o Hai khng sinh c tc dng hip ng nu ch s FIC 0.5

o Hai khng sinh c tc dng i khng nu ch s FIC > 4

o Hai khng sinh phi hp khng c tc dng khc bit nu ch s FIC 4 v > 0.5




Nhng trng hp tc dng i khng yu c th khng pht hin c v

vng c ch nm di vng bt cho ca hai thanh E-test.

Sai st c th gp khi:



- Phi tin hnh lm li khi thy cc hin tng nh trn.

31

10. Vi khun k kh nui cy v nh danh

I. MC CH V NGUYN L

1. Mc ch

Pht hin v nh danh vi khun k kh gy bnh bng phng php nui

cy kinh in.

2. Nguyn l

Vi khun c nh danh da vo c im nui cy, mt s tnh cht

chuyn ha, cc c im v hnh thi hc v c th kt hp vi tnh cht khng

nguyn.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1. Trang thit b



- H thng my nui cy vi khun k kh




2 Lam knh Ci 2,000

3 Que cy Ci 2,000






9 Cn ty 95 ml 10,000

10 Lugol ml 5,000

32


12 Mi trng nui cy k kh chai 1,000

13 Mi trng thch mu thng a 2,000

14 Mi trng thch mu k kh a 2,000

15 B gi ng API Test 1,000

16 Bnh kh trn (tnh theo gi) Gi 168,000


18 Bng Kg 0,001

19 Cn 90 (v sinh dng c) ml 10,000

20 n cn Ci 0,0001

21 Panh Ci 0,0001



24 M Ci 0,020


26 Gng tay i 3,000


28 Qun o bo h bo h B 0,001




32 Bt bi Ci 0,010

33 Bt la Ci 0,010








* Ghi ch:

33







3. Bnh phm

Tt c cc loi bnh phm c ch nh nui cy tm vi khun k kh gy bnh.

4. Phiu xt nghim


III. CC BC TIN HNH


1. Ly bnh phm


2. Tin hnh k thut



- m qua m


- Nhum soi, th nghim cc th nghim sinh vt ha hc n gin v nh danh bng cc b sinh vt ha hc chuyn dng.

- Tt c cc bc tin hnh v m u c thc hin bn trong h thng my nui cy vi khun k kh nhm m bo iu kin mi trng k kh.

IV. NHN NH KT QU




Quy trnh ny ch p dng tm vi khun k kh bt buc v k kh ty tin.

Kt qu m tnh khng c ngha l khng c vi khun gy bnh trong bnh

phm m l khng tm thy cn nguyn vi khun gy bnh c th phn lp c

bng quy trnh nui cy ny.

Tt c cc thao tc k thut phi din ra bn trong h thng my to mi

trng k kh m bo iu kin nui cy thch hp cho vi khun k kh mc.

Kh trng s dng nui cy vi khun phi m bo ng nng .


34

11. Vi khun khng nh

I. MC CH V NGUYN L

1. Mc ch

Pht hin vi khun gy bnh bng phng php nui cy, nh danh trn

h thng t ng.

2. Nguyn l


chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1. Trang thit b


- T m thng

- T m CO2


- My o c

- L hp t





2 Lam knh Ci 2,000

3 Que cy Ci 1,000




35

7 Thuc nhum Fuchsin ml 5,000

8 Thuc nhum tm Gentian ml 5,000

9 Cn ty 95 ml 10,000

10 Lugol ml 5,000






16 Bng Kg 0,001

17 Cn 90 (v sinh dng c) ml 10,000

18 n cn Ci 0,0001

19 Panh Ci 0,0001



22 M Ci 0,020


24 Gng tay i 3,000


26 Qun o bo h B 0,001




30 Bt bi Ci 0,010

31 Bt la Ci 0,010








36


* Ghi ch:







3. Bnh phm


4. Phiu xt nghim


III. CC BC TIN HNH


1. Ly bnh phm


2. Tin hnh k thut



- m qua m



IV. NHN NH KT QU











37

12. Vi khun nh danh PCR

I. MC CH V NGUYN L

1. Mc ch

Chn on nhanh vi khun trc tip t bnh phm.

2. Nguyn l

Xc nh s c mt ca gene c trng cho vi khun bng k thut PCR.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1. Trang thit b

- T an ton Sinh hc cp 2

- My PCR

- My in di

- My c in di

- My ly tm 25000 x g

- My nhit

- My vortex

- My ly tm dng cho tube 0,2 ml

- Micropipette

- T m su (-200C)

- B lu in.


nh mc sinh phm v vt t tiu hao cho 2 mu/ln thc hin (VD)


1 Bng kg 0,001

2 Cn ml 1,000


38

4 Panh Ci 0,0001


6 Hp vn chuyn bnh phm Test 0,001


8 Gng khng c bt (DNase-RNase free) Ci 0,500


10 Khu hao sinh phm cho kim tra li Test 1,350

11 Kit tch chit DNA Test 2,350

12 DNA marker B 1,000

13 Primer 1 ml 0,0001

14 Primer 2 ml 0,0001

15 Ependoff 1,7ml Tube 3,000

16 Ependoff 0,2ml Tube 1,000

17 u cn 10 l c lc Ci 1,000

18 u cn 30 l Ci 1,200

19 u cn 200 l c lc Ci 2,200

20 u cn 1 ml c lc Ci 3,200

21 Ethanol BDH ml 0,500

22 Water-DEPC Treated ml 2,000





27 Bt bi Ci 0,010

28 M Ci 0,020


30 Gng tay i 0,100


32 Qun o bo h B 0,005




39

36 Khn lau tay ci 0,010

37 QC (nu thc hin) * 0,100


* Ghi ch:







3. Bnh phm

Vi khun thun v bnh phm phn lp

4. Phiu xt nghim


III. CC BC TIN HNH


1. Ly bnh phm


2. Tin hnh k thut

- Tch chit DNA tng s

- Thc hin PCR

- in di kim tra sn phm

- nh gi v kt lun

IV. NHN NH KT QU

Sn phm PCR phi c mt bng c hiu duy nht, r nt, khng b t gy

v c kch thc tng ng vi on gene ch cn khuch i.


Khi kt qu khi chng m v/hoc chng dng khng hp l, cn phi

kim tra li qu trnh tch chit DNA tng s, cht lng primers v master

mix, sau thc hin li xt nghim.

40

13. Vi khun nh danh gii trnh t gene

I. NGUYN L

1. Mc ch

nh danh mi cn nguyn vi khun gy bnh n mc chi v hoc

loi.

2. Nguyn l

nh danh chnh xc cc loi vi khun da trn trnh t nucleotide c

trng ca gene m ha cho 16S rRNA ca vi khun bng k thut gii trnh t

gene.

II. CHUN B

1.Ngi thc hin



2. Phng tin, ha cht


2.1. Trang thit b


- My PCR

- My c in di

- My gii trnh t gen


- My nhit

- My vortex


- Micropipette

- B lu in




1 Bng kg 0,001

2 Cn ml 1,000

41


4 Panh Ci 0,0001









13 Primer 1 (b primer c hiu) ml 0,0001




17 ng Eppendorf 1,5 ml Tube 3,000


19 u cn 10 l c lc Ci 1,000

20 u cn 30 l Ci 1,200

21 u cn 200 l c lc Ci 2,200

22 u cn 1 ml c lc Ci 3,200







29 Bt bi Ci 0,010

30 M Ci 0,020


32 Gng tay i 0,100


34 Qun o bo h B 0,005

42




38 Khn lau tay ci 0,010

39 QC (nu thc hin) * 0,100


* Ghi ch:







3. Bnh phm

Chng vi khun nui cy thun.

4. Phiu xt nghim


III. CC BC TIN HNH


1. Ly bnh phm


2. Tin hnh k thut

- Tch chit DNA tng s

- Thc hin PCR gene 16S ARN ribosome

- in di kim tra sn phm

- Gii trnh t gene

- Kim tra tng ng DNA

IV. NHN NH KT QU

Sn phm PCR phi c mt bng c hiu duy nht, r nt v khng b

t gy. Trnh t DNA ca gene ch c so snh vi cc trnh t ca gene m

ha cho 16S rRNA c trn Genebank nh danh cn nguyn vi khun cn

xc nh. Trnh t nucleotide phi c tng ng 97 % mi c th kt lun

c n mc chi v/hoc loi.

43


- Trong trng hp khng c sn phm PCR, cn phi kim tra li qu

trnh tch chit DNA tng s, cht lng primers v master mix, v thc hin

li.

- Nu trnh t DNA b nhiu cn phi kim tra li c hiu ca sn

phm PCR hoc qu trnh chy PCR sequencing b nhim cho.

44

14. Vi khun khng thuc PCR

I. MC CH V NGUYN L

1. Mc ch

Pht hin c tnh khng thuc ca vi khun thng qua xc nh s c

mt ca gene khng thuc vi khun.

2. Nguyn l

S dng cp mi c trnh t c hiu xc nh s c mt ca gene

khng thuc trong vi khun gy bnh bng k thut PCR.

II. CHUN B

1. Ngi thc hin



2. Phng tin, ha cht


2.1. Trang thit b


- My PCR

- My c in di


- My nhit

- My vortex


- Micropipette

- B lu in.




1 Bng kg 0,001

2 Cn ml 1,000


4 Panh Ci 0,0001


45








13 Primer 1 ml 0,0001

14 Primer 2 ml 0,0001



17 u cn 10 l c lc Ci 1,000

18 u cn 30 l Ci 1,200

19 u cn 200 l c lc Ci 2,200

20 u cn 1 ml c lc Ci 3,200







27 Bt bi Ci 0,010

28 M Ci 0,020


30 Gng tay i 0,100


32 Qun o bo h B 0,005


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