Sensitive Mie scattering immunoagglutination assay of porcine reproductive and respiratory syndrome virus (PRRSV) from lung tissue samples in a microfluidic chip

Jae Young Song, Chang Hee Lee, Eun Jin Choi, Keesung Kim, Jeong-Yeol Yoon

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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. Antibodies against PRRSV were conjugated to the surface of highly carboxylated polystyrene microparticles (diameter=920nm) 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 380nm incident beam using microcallipered, proximity fiber optics. For comparison, multi-well experiments were also performed using the same optical detection setup. The detection limit was determined to be 10 -3TCID 50ml -1 for PRRSV dissolved in PBS, while those of previous RT-PCR studies for PRRSV were 10 1TCID 50ml -1 (conventional assays) or <1TCID 50ml -1 (quantitative real-time assays). Mie scattering 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 5min. A strong correlation could be found between RT-PCR and this method for the lung tissue samples, even though their respective detection mechanisms are different fundamentally (nucleic acids for RT-PCR 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. Specificity studies revealed the presence of Type II PRRSV and non-presence of Type I PRRSV and that the microfluidic chip assay could detect Type II North American strain of PRRSV for the animals tested. 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.

Original languageEnglish (US)
Pages (from-to)31-38
Number of pages8
JournalJournal of Virological Methods
Volume178
Issue number1-2
DOIs
StatePublished - Dec 2011

Fingerprint

Porcine respiratory and reproductive syndrome virus
Microfluidics
Lung
Polymerase Chain Reaction
Optical Phenomena
Light
Antigens
Antibodies
Polystyrenes
Domestic Animals
Computer Systems
Immunoassay
Nucleic Acids
Limit of Detection
Swine
Viruses
Sensitivity and Specificity

Keywords

  • Lab-on-a-chip
  • Latex agglutination test
  • Microfluidic immunosensor
  • One-step RT-PCR
  • PRRSV

ASJC Scopus subject areas

  • Virology

Cite this

Sensitive Mie scattering immunoagglutination assay of porcine reproductive and respiratory syndrome virus (PRRSV) from lung tissue samples in a microfluidic chip. / Song, Jae Young; Lee, Chang Hee; Choi, Eun Jin; Kim, Keesung; Yoon, Jeong-Yeol.

In: Journal of Virological Methods, Vol. 178, No. 1-2, 12.2011, p. 31-38.

Research output: Contribution to journalArticle

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abstract = "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. Antibodies against PRRSV were conjugated to the surface of highly carboxylated polystyrene microparticles (diameter=920nm) 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 380nm incident beam using microcallipered, proximity fiber optics. For comparison, multi-well experiments were also performed using the same optical detection setup. The detection limit was determined to be 10 -3TCID 50ml -1 for PRRSV dissolved in PBS, while those of previous RT-PCR studies for PRRSV were 10 1TCID 50ml -1 (conventional assays) or <1TCID 50ml -1 (quantitative real-time assays). Mie scattering 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 5min. A strong correlation could be found between RT-PCR and this method for the lung tissue samples, even though their respective detection mechanisms are different fundamentally (nucleic acids for RT-PCR 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. Specificity studies revealed the presence of Type II PRRSV and non-presence of Type I PRRSV and that the microfluidic chip assay could detect Type II North American strain of PRRSV for the animals tested. 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.",
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