猴T淋巴细胞趋向性病毒1型双抗原夹心ELISA检测试剂盒的研制

Development of Double-antigen Sandwich ELISA Kit of Simian T-lymphotropic Virus Type-I

目的研制灵敏度和特异性高的检测实验猴血清中T淋巴细胞趋向性病毒-1型(STLV-1型)抗体的双抗原夹心ELISA(dsELISA)检测试剂盒。方法采用经原核表达系统表达并纯化的人T淋巴细胞白血病病毒-1型(HTLV-1型)的Env蛋白作为包被用抗原,建立了检测STLV-1的dsELISA诊断方法。通过优化反应条件和筛选试剂,确定了dsELISA诊断试剂盒的相关条件,并经敏感性、特异性和重复性试验考查该试剂盒质量。结果试剂盒特异性好,批内重复试验变异系数(CV)<7%,批间重复试验CV<10%。对200份猴血清进行随机检测,与国际公认的诊断试剂盒(美国BioReliance公司)的符合率为97%。结论本试剂盒可初步应用于临床上实验猴STLV-1型抗体的检测。

Objective To develope a sensitive and specific double-antigen sandwich ELISA（dsELISA） kit for detection of simian T-lymphotropic Virus type 1 （STLV-1） serum antibody. Methods The envelope protein of human T-lymphotropic virus type 1 （HTLV-1）expressed in E. coil was used as detective antigen in dsELISA kit. The dsELISA method was established by optimization reaction conditions and selection of reagents. The sensitivity, specificity and reproducibility of the kit were also studied in this paper. Results The kit has good specificity and reproducibility. The variant coefficients of the OD value within assay and between assay were less than 7% and 10%, respectively. 200 samples of monkey sera were tested with this kit, and results showed that it has 97 % identification with the standard kit（ BioReliance company, USA）. Conclusion The kit developed in this paper has several advantages compared with standard kit, and can be applied in STLV-1 antibody detection and research.