Abstract: Objective In order to survey and investigate the epidemic character of simian T-cell lymphotropic virus type 1(STLV-1)transmission among wild and experimental monkeys, method of recombinase polymerase amplification (RPA) and fluorescent RPA to detect STLV-1 were established. Method Analysis conservative sequence of STLV-1 gag polyprotein genes separated from different countries and areas, RPA primers and fluorescent probe were designed and RPA methedes were tested by specificity, sensitivity and stability. Result Compared with other simian viruses, new RPA and fluorescent RPA mothedes had highly specific to STLV-1. The limit of detection of RPA and fluorescent RPA mothedes is similar with PCR method. The stability of RPA and fluorescent RPA was comfirmed by detection of 30 postive and negative STLV-1 samples. Conclusion The RPA and fluorescent RPA mothedes to detect STLV-1 were established and proved to be specific with high sensitivity and stability, which could be applied to survey STLV-1 infection among simian.

Key words: simian, simian T-cell lymphotropic virus type 1, recombinase polymerase amplification(RPA), fluorescent RPA

摘要： 目的 为了提高口岸对进出境野生及实验用灵长类动物猴T淋巴细胞趋向性病毒l型(Simian T-cell lymphotropic virus type 1,STLV-1)感染情况的监测和流行病学调查,本研究建立了重组酶聚合酶扩增(RPA)和荧光RPA检测STLV-1的方法.方法 使用软件分析不同国家和地区分离的STLV-1的gag蛋白(gag polyprotein)基因序列,设计RPA引物和探针,建立RPA和荧光RPA检测STLV-1的方法,并对方法的特异性、敏感性和稳定性进行验证.结果 通过将猴其它特定病原体与STLV-1对比检测,证实建立的检测方法 具有良好的特异性;通过敏感性试验,证实建立的RPA和荧光RPA方法检测下限与PCR一致;通过对30份STLV-1阳性和阴性核酸样本的检测,证实建立的RPA和荧光RPA方法,具有PCR方法一样的稳定性和可靠性.结论 本研究建立的RPA和荧光RPA检测STLV-1的方法具有良好的特异性、敏感性和可靠性,可应用于STLV-1的监测和流行病学调查.

关键词: 猴, 猴T淋巴, 细胞趋向性病毒Ⅰ型, RPA技术, 荧光RPA