ismm2015 kagiyama - 名古屋大学€¦ · -20-15-10-5 0 02 4681012 oscillation circuit...

-20 -15 -10 -5 0 0 2 4 6 8 10 12 Oscillation Circuit Microfluidic Channel PDMS 1 Nucleoprotein(NP) RNA PNA Microfluidic channel QCM PDMS 2 NP antibody Gold colloid QCM Microfluidic chip with QCM for detection of influenza virus subtype ○S. Kagiyama 1 T. Masuda 1 K. Kaihatsu 2 N. Kato 2 F. Arai 1 1 Nagoya University, JAPAN 2 Osaka University, JAPAN Experiment Sauerbrey’s equation Conclusions Power Supply Microfluidic channel Oscillation Circuit Frequency Counter Signal Output Viral RNA amount [pfu] Condition 1 5×10 3 Condition 2 10 4 RNA only(10 6 pfu/mL) Gold colloids only Condition 1 Condition 2 Contact person： Taisuke Masuda E-mail: [email protected], URL: http://www.biorobotics.mech.nagoya-u.ac.jp/ Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan Dept. of Micro-Nano Systems Engineering, Nagoya University TEL: 052-789-5026, FAX : 052-789-5027 Acknowledgements： This work was supported in part by the Nano-Macro Materials, Devices and System Research Alliance Project from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT). Reference [1] M.Niimi et al, “Virus purification and enrichment by hydroxyapatite chromatography on a chip”, volume 201, 2014, P185-190 Sensors and Actuators B [2] K.Kaihatsu et al, ”Sequence-Specific and Visual Identification of the Influenza Virus NS Gene by Azobenzene-Tethered Bis-Peptide Nucleic Acid” 2013, PLOS ONE [3] S.Kagiyama et al, ”Detection of influenza virus subtype using quartz crystal microbalance” 2014, Micro-Nano Mechatronics and Human Science (MHS) Virus purification by Hydroxyapatite Viral DNA/RNA extraction by Sillica Purpose Establish the method to detect viral RNA. Influenza Dengue fever etc… Φ3 mm Thickness : 0.1 mm Fundamental frequency f 0 540 times heavier Mass [g/piece] Size [nm] Viral RNA 1.2×10 -18 40~80 Gold colloid 6.5×10 -16 Φ 40 1. Viral RNA captured specifically by Peptide Nucleic Acid (PNA) . 2. Gold colloids captured to increase sensing mass . RNA PNA K. Kaihatsu et al., 2013, PLOS ONE Oscillation circuit Thickness slip vibration + + Quartz Crystal Microblance Peptide nucleic acid Sensitizer (Gold colloids) Sensitizer Gold colloid RNA Height of channel : 200 μm PDMS 8.4 mm 3.4 mm Infectious disease Background Conventional technique Concept Design QCM Microfluidic channel Quartz Crystal Microbalance 1 : Introduced viral RNA (Condition 1, 2). 2 : Introduced Gold colloids . Δ Frequency [Hz] Time [min] Detection of viral RNA Setting QCM Need simple and accurate Diagnosis Previous method Rapid diagnosis kit 10 mm Microfluidic channel QCM Oscillation circuit Connector ○Rapidly ×Specifically Rapid diagnosis kit (BD DirectigenTM FluA) Our chip Detection limit 10 3 ~ 10 4 5×10 3 Detect subtype × ○ ○ Detected viral RNA using QCM by increasing sensing mass. ○ In our future work, we will integrate all the functions in one chip.

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Page 1: ISMM2015 Kagiyama - 名古屋大学€¦ · -20-15-10-5 0 02 4681012 Oscillation Circuit Microfluidic Channel 1 PDMS Nucleoprotein(NP) RNA PNA Microfluidic channel QCM 2 PDMS Gold

-20

-15

-10

-5

0 2 4 6 8 10 12

OscillationCircuit

MicrofluidicChannel

PDMS1

Nucleoprotein(NP)RNA

PNAMicrofluidic

channel

QCM

PDMS2

NP antibodyGold colloid

QCM

Microfluidic chip with QCMfor detection of influenza virus subtype

○S. Kagiyama1 T. Masuda1 K. Kaihatsu2 N. Kato2 F. Arai11Nagoya University, JAPAN

2Osaka University, JAPAN

ExperimentSauerbrey’s equation

Conclusions

PowerSupply

Microfluidicchannel

OscillationCircuit

FrequencyCounter

Signal Output

Viral RNA amount [pfu]Condition 1 5×103

Condition 2 104

RNA only(106 pfu/mL)

Gold colloids only

Condition 1

Condition 2

Contact person： Taisuke Masuda E-mail: [email protected], URL: http://www.biorobotics.mech.nagoya-u.ac.jp/Furo-cho, Chikusa-ku, Nagoya 464-8603, JapanDept. of Micro-Nano Systems Engineering, Nagoya UniversityTEL: 052-789-5026, FAX : 052-789-5027

Acknowledgements：This work was supported in part by the Nano-Macro Materials, Devices and SystemResearch Alliance Project from the Ministry of Education, Culture, Sports, Scienceand Technology of Japan (MEXT).

Reference [1] M.Niimi et al, “Virus purification and enrichment by hydroxyapatite chromatography on a chip”, volume 201, 2014, P185-190 Sensors and Actuators B[2] K.Kaihatsu et al, ”Sequence-Specific and Visual Identification of the Influenza Virus NS Gene by Azobenzene-Tethered Bis-Peptide Nucleic Acid” 2013, PLOS ONE [3] S.Kagiyama et al, ”Detection of influenza virus subtype using quartz crystal microbalance” 2014, Micro-Nano Mechatronics and Human Science (MHS)

Virus purification by Hydroxyapatite Viral DNA/RNA extraction by Sillica

PurposeEstablish the method to detect viral RNA.

Influenza Dengue fever etc…

Φ3 mm

Thickness : 0.1 mm

Fundamental frequency f0

540 times heavier

Mass [g/piece] Size [nm]

Viral RNA 1.2×10-18 40~80

Gold colloid 6.5×10-16 Φ 40

1. Viral RNA captured specifically by Peptide Nucleic Acid (PNA) .2. Gold colloids captured to increase sensing mass .

RNA