Proper Collection, HandlingProper Collection, Handlingand Storage of Respiratoryand Storage of Respiratory

SpecimensSpecimensSpecimensSpecimens

National Influenza CenterNational Influenza CenterResearch Institute for Tropical MedicineResearch Institute for Tropical Medicine

AlabangAlabang, , MuntinlupaMuntinlupa CityCity

1. Preferred RespiratorySpecimens:

1.1 nasopharyngeal swab and oropharyngeal swab

1.2 nasopharyngeal aspirate 1.2 nasopharyngeal aspirate 1.3 nasal wash/aspirate

Supplies/Equipment NeededSupplies/Equipment Needed Case Report Form (CRF)Case Report Form (CRF) Virus Transport Media (VTM)Virus Transport Media (VTM) Nasopharyngeal swab, Sterile Dacron/Rayon swab Nasopharyngeal swab, Sterile Dacron/Rayon swab

with pliable shaftwith pliable shaft Oropharyngeal swab, Sterile Dacron/Rayon swab Oropharyngeal swab, Sterile Dacron/Rayon swab

with plastic shaftwith plastic shaft Sterile tongue depressorSterile tongue depressor Sterile tongue depressorSterile tongue depressor Thermo box with 4Thermo box with 4--6 frozen ice packs6 frozen ice packs Test Tube rackTest Tube rack Zip Lock bagsZip Lock bags Parafilm Parafilm Masking tapeMasking tape Permanent tube markerPermanent tube marker PPE (gloves, mask, goggles and lab.gown)PPE (gloves, mask, goggles and lab.gown) RefrigeratorRefrigerator

Details of ProcedureDetails of Procedure

1.1. PrePre-- specimen collectionspecimen collection

1.1. Take out the VTMs from the freezer (1.1. Take out the VTMs from the freezer (--2020ºC) where ºC) where 1.1. Take out the VTMs from the freezer (1.1. Take out the VTMs from the freezer (--2020ºC) where ºC) where they are stored.they are stored.

1.21.2 Frozen VTMs should be thawed just before use. If Frozen VTMs should be thawed just before use. If the collection site is far from a refrigerator, have a the collection site is far from a refrigerator, have a thermo box with 4thermo box with 4--6 frozen ice packs at hand to 6 frozen ice packs at hand to maintain a refrigerated temperature during maintain a refrigerated temperature during

collection.collection.

1.3 1.3 Check VTM for turbidity. The medium Check VTM for turbidity. The medium should be clear and pinkish. should be clear and pinkish. Tap the tube to mix contents.Tap the tube to mix contents.

1.4 Check also the integrity of the swab and 1.4 Check also the integrity of the swab and tongue depressor pouch to ensure sterility. Do not use swabs or tongue depressor which have been opened.

2. Case Investigation and Documentation2. Case Investigation and Documentation

2.1. The physician investigates the case and 2.1. The physician investigates the case and fills out a fills out a Case Report FormCase Report Form ( done before ( done before the collection of specimen) from suspected the collection of specimen) from suspected cases. cases.

2.2. The physician/nurse/med. tech. will collect 2.2. The physician/nurse/med. tech. will collect

specimens from suspected casesspecimens from suspected cases

3. Specimen collection3. Specimen collection

3.1 Label the VTM tube with the patient’s Full Name and date of collection. The information on the label must be legible and should match the information on the Case Report Form. the information on the Case Report Form. Label must remain attached under all conditions of storage and transport.

IMPORTANTIMPORTANT

UseUse onlyonly sterilesterile DacronDacron oror RayonRayon--tiptipswabsswabs withwith plasticplastic shaftshaft .. DoDo notnot useusecottoncotton swabswab withwith woodenwooden shaftshaft asas thisthiscottoncotton swabswab withwith woodenwooden shaftshaft asas thisthiswillwill inactivateinactivate somesome virusesviruses andand willwill alsoalsoinhibi tinhibi t detect iondetect ion inin PCRPCR testingtest ing..

A. Nasopharyngeal swab

3.2 Using a flexible Dacron or Rayon tip swab, measure from the base of the nostril towards the auditory towards the auditory pit. Divide the length into half in order to know into what extent will be inserted into the nostril (usually 5–6 cm in adults) to ensure that it reaches the posterior pharynx.

A.Nasopharyngeal swab

3.3. With the patient seated, tilt the head slightly backwards. slightly backwards. Insert the swab into the nostril parallel to the palate

A.Nasopharyngeal swab

3.4 Rotate swab applying a little force taking large quantities of large quantities of mucosa.

3.5 Repeat procedure in the other nostril using the same swab (if possible)

Nasopharyngeal SwabNasopharyngeal Swab

3.6 Place the nasopharyngeal swab immediately in the VTM tube to avoid drying of the swab.

3.7 Break/cut with scissors the end of the swab that sticks out of the tube and close the tube.

Oropahryngeal swab(Posterior pharyngeal swab)

3.8 With gloved hands hold down the tongue with a sterile tongue depressor.depressor.

3.9 Have the patient say "aahh“to elevate the uvula.

Swab held correctly

Swab held incorrectly

Correctly held swab can slide out of the way.

Swab can injure patient.

Oropahryngeal swab(Posterior pharyngeal swab)

4.0 Use a sweeping motion to swab the posterior pharyngeal wall and tonsilar pillars. Apply a little force, taking large quantities of mucosa.taking large quantities of mucosa.

4.1 Avoid swabbing the soft palate and do not touch the tongue with the swab tip.

(N.B. This procedure can induce the gag reflex);

4.2 Place the oropharyngeal swab immediately in the same VTM tubewith the nasopharyngeal swab.

4.3 Break/cut with scissors the end of the swab that sticks out of the tube the swab that sticks out of the tube and close the tube tightly. Secure the cap with parafilm to prevent leakage during transport.

4.4 Wrap VTM tubes with specimens in tissue paper or any absorbent material;place upright in a separate 50 ml centrifuge separate 50 ml centrifuge tube or any leak/puncture proof container. Place the 50 ml tube or any container in a resealable plastic bag (ZiplockTM).

4.5 Store inside the refrigerator (2-8ºC)/thermobox with ice packs while awaiting transport.transport.

4.6 Transport to RITM immediately or should reach RITM within 72 hours after collection