基于核酸适体LYGV1c的酶联吸附法检测石斑鱼虹彩病毒感染OA北大核心CHSSCDCSTPCD

[Objective]To develop the aptamer LYGV1c-based enzyme-linked apta-sorbent assay(LYGV1c-ELASA)for accurate and rapid detection of Singapore grouper iridovirus Guangxi strain(SGIV-Gx),and to provide theoretical support for the development of rapid detection system technology in aquaculture.[Method]An aptamer enzyme-linked adsorption method was developed based on biotin-labeled LYGV1c.The specificity,sensitivity and stability of bio-LYGV1c were analyzed in SGIV-Gx infection.Hybrid Grouper(Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂)were infected by SGIV-Gx,and the relative expression of MCP of SGIV-Gx was determined by fluorescence quantitative PCP(qPCR),the results of qPCR were compared with that of LYGV1c-ELASA to verify the reliability of ELASA.[Result]LYGV1c-ELASA could detect SGIV infection with high specificity,and the working concentration was 500 nmol/L,incubation was 20 min,the suitable binding temperature was 4‒28℃,and the detection limit was 5×103 cells/mL.The results of infection test showed that the OD450 value of SGIV-Gx increased with the increase of virus concentration in vivo.The qPCR result was the same as LYGV1c-ELASA detection results in vivo.[Conclusion]LYGV1c-ELASA technology can be used for the detection of SGIV-Gx in vitro and in vivo.LYGV1c-ELASA is a technique for rapid diagnosis and real-time monitor of SGIV-Gx infection.