基于上转化发光免疫层析技术建立发热伴血小板减少综合征病毒总抗体快速检测方法

Rapid detection of severe fever with thrombocytopenia syndrome virus(SFTSV)total antibodies by up-converting phosphor technology(UPT)-based lateral-flow assay

目的基于上转化发光(UPT)免疫层析技术,建立发热伴血小板减少综合征病毒(SFTSV)总抗体的现场快速检测方法。方法将SFTSV重组NP蛋白与上转化发光颗粒(UCP)偶联,制备UCP-NP免疫层析试纸条,评价该试纸条检测SFTSV总抗体的灵敏性、特异性和稳定性,并检测SFTSV血清254份,与酶联免疫法(ELISA)比较。结果该方法可在15min内完成SFTSV总抗体检测,可检测1∶500稀释度的SFTSV阳性血清,与其他出血热病毒无交叉反应,加样14d内稳定性较高。UPT免疫层析法与ELISA法检测临床血清样品一致性极高(Kappa=0.967),约登指数为0.973。结论建立了基于UPT免疫层析技术的SFTSV总抗体快速检测方法,该方法灵敏、特异,且操作简便、快速,结果稳定,适合在基层门诊和体检现场推广。

Objective To develop a method for rapid and on-site detection of severe fever with thrombocytopenia syndrome virus （SFTSV） total antibodies by up-converting phosphor technology （UPT） -based lateral-flow assay. Methods SFTSV re- combinant NP protein was conjugated to up-converting phosphor particles （UCP）, UPT-NP analysis strips were assembled. The sensitivity, specificity and stability of UPT method to detect total antibodies were evaluated; 254 clinical sera were tested by UPT method and ELISA to study the consistence of above 2 methods. Results UPT method can analyze total antibodies a- gainst SFTSV in 15 minutes, with the detection limit of 1：500 dilution of original positive SFTSV sera samples; no cross reac- tion with other hemorrhagic fever virus were detected. UPT method exhibited high stability in 14 days upon sample loading. The results of UPT method were highly consistent with ELISA （Kappa=0. 967）, with Youden＇s index of 0. 973. Conclusion Rapid detection of SFTSV total antibodies by UPT-based lateral-flow assay was developed successfully, which was sensitive , specific , rapid ,stable and easy to operate. It is especially suitable to be promoted in local clinics and on-site physical examina- tions.