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축사 내 병원체 검출용 랩온어칩 개발에 관한 연구(Development of lab-
학
과제명
on-a-chip to detect infectious agents within barns)
술
발
Detection of porcine reproductive and respiratory syndrome virus using Mie
발표제목
scattering immunoagglutination assay in a microfluidic chip
표
?
발표자
최은진
국
발표처
국제동물전문가학회(CRWAD)
발표시기
2011년 12월
외
?
수록잡지
세계 동물질병전문가회의 (CRWAD) 초록집
Abstract：
Purpose: A microfluidic immunosensor utilizing Mie scattering immunoaggultination assay
was developed for rapid and sensitive detection of porcine reproductive and respiratory
syndrome virus (PRRSV) from lung tissue samples of domesticated pigs.
Methods: Antibodies against PRRSV were conjugated to the surface of highly carboxylated
polystyrene microparticles (diameter = 920 nm) and mixed with the diluted PRRSV tissue
samples in a Y-shaped microchannel. Antibody-antigen binding induced microparticle
immunoagglutination, which was detected by measuring the forward 45° light scattering of
380 nm incident beam using microcallipered, proximity fiber optics. For comparison, multi-well
experiments were also performed using the same optical detection setup.
Results: The detection limit was determined to be 10-3 TCID50㎖-1 for PRRSV dissolved in
PBS, while those of previous RT-PCR studies for PRRSV were 101 TCID50㎖-1
(conventional assays) or
simulations were able to predict the shape of the PRRSV standard curve, indicating that any
non-linearity of the standard curve can be interpreted purely as an optical phenomenon. Each
assay took less than 5 min. A strong correlation could be found between RT-PCR and this
method for the lung tissue samples, even though their respective detection mechanisms are
fundamentally different (nucleic acids for RTPCR and virus antigens for light scattering
immunoagglutination assay). Several different dilution factors were also tested for tissue
samples, and 1/10 and 1/100 were found to be usable. If the microfluidic chips are used only
once (i.e. without re-using them), both superior sensitivity and satisfactory specificity can be
demonstrated.
Conclusions: This work demonstrates the potential of the Mie scattering immunoassay on a
microfluidic chip towards real-time detection system for viral pathogens in domesticated
animals.
799
학
과제명
on-a-chip to detect infectious agents within barns)
술
발
Detection of porcine reproductive and respiratory syndrome virus using Mie
발표제목
scattering immunoagglutination assay in a microfluidic chip
표
?
발표자
최은진
국
발표처
국제동물전문가학회(CRWAD)
발표시기
2011년 12월
외
?
수록잡지
세계 동물질병전문가회의 (CRWAD) 초록집
Abstract：
Purpose: A microfluidic immunosensor utilizing Mie scattering immunoaggultination assay
was developed for rapid and sensitive detection of porcine reproductive and respiratory
syndrome virus (PRRSV) from lung tissue samples of domesticated pigs.
Methods: Antibodies against PRRSV were conjugated to the surface of highly carboxylated
polystyrene microparticles (diameter = 920 nm) and mixed with the diluted PRRSV tissue
samples in a Y-shaped microchannel. Antibody-antigen binding induced microparticle
immunoagglutination, which was detected by measuring the forward 45° light scattering of
380 nm incident beam using microcallipered, proximity fiber optics. For comparison, multi-well
experiments were also performed using the same optical detection setup.
Results: The detection limit was determined to be 10-3 TCID50㎖-1 for PRRSV dissolved in
PBS, while those of previous RT-PCR studies for PRRSV were 101 TCID50㎖-1
(conventional assays) or
simulations were able to predict the shape of the PRRSV standard curve, indicating that any
non-linearity of the standard curve can be interpreted purely as an optical phenomenon. Each
assay took less than 5 min. A strong correlation could be found between RT-PCR and this
method for the lung tissue samples, even though their respective detection mechanisms are
fundamentally different (nucleic acids for RTPCR and virus antigens for light scattering
immunoagglutination assay). Several different dilution factors were also tested for tissue
samples, and 1/10 and 1/100 were found to be usable. If the microfluidic chips are used only
once (i.e. without re-using them), both superior sensitivity and satisfactory specificity can be
demonstrated.
Conclusions: This work demonstrates the potential of the Mie scattering immunoassay on a
microfluidic chip towards real-time detection system for viral pathogens in domesticated
animals.
799
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