Detection of VR-2332 Strain of Porcine Reproductive and Respiratory Syndrome Virus Type II Using an Aptamer-Based Sandwich-Type Assay

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• 作者：Su Jin Lee ; Young Seop Kwon ; Ji-eun Lee ; Eun-Jin Choi ; Chang-Hee Lee ; Jae-Young Song ; Man Bock Gu
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：January 2, 2013
• 年：2013
• 卷：85
• 期：1
• 页码：66-74
• 全文大小：515K
• 年卷期：v.85,no.1(January 2, 2013)
• ISSN：1520-6882

文摘

Porcine reproductive and respiratory syndrome virus (PRRSV) causes porcine reproductive and respiratory syndrome disease (PRRS), a disease that has a significant and economic impact on the swine industry. In this study, single-stranded DNA (ssDNA) aptamers with high specificity and affinity against VR-2332 strain of PRRSV type II were successfully obtained. Of 19 candidates, the LB32 aptamer was found to be the most specific and sensitive to VR-2332 strain according to an aptamer-based surface plasmon resonance (SPR) analysis. The detection of VR-2332 of PRRSV type II was successfully accomplished using the enzyme-linked antibody-aptamer sandwich (ELAAS) method. The detection limit of ELAAS was 4.8 脳 10⁰ TCID₅₀/mL that is comparable to some of the previous reports of the PCR-based detection but does not require any complicated equipment or extra costs. Moreover, this ELAAS-based PRRSV detection showed similar sensitivity for both the VR-2332 samples spiked in diluted swine serum and in buffer. Therefore, this VR-2332 strain-specific aptamer and its assay method with high specificity can be used as an alternative method for the fast and precise detection of PRRSV.