title: Detection of VR-2332 strain of porcine reproductive and respiratory syndrome virus type II using an aptamer-based sandwich-type assay.
abstract: 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 × 10(0) TCID(50)/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.
authors.affiliation: College of Life Sciences and Biotechnology, Korea University, Anam-Dong, Seongbuk-Gu, Seoul, Republic of Korea.
authors.forename: Su Jin
authors.initials: SJ
authors.lastname: Lee
authors.forename: Young Seop
authors.initials: YS
authors.lastname: Kwon
authors.forename: Ji-eun
authors.initials: JE
authors.lastname: Lee
authors.forename: Eun-Jin
authors.initials: EJ
authors.lastname: Choi
authors.forename: Chang-Hee
authors.initials: CH
authors.lastname: Lee
authors.forename: Jae-Young
authors.initials: JY
authors.lastname: Song
authors.forename: Man Bock
authors.initials: MB
authors.lastname: Gu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000737:chemistry; D007163:Immunosorbent Techniques / Q000295:instrumentation; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000032:analysis* / Q000097:blood; D020349:Surface Plasmon Resonance; D013552:Swine
chemical_list: D000911:Antibodies, Monoclonal; D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D012367:RNA, Viral