重组人类T淋巴细胞白血病病毒抗原及双抗原夹心法ELISA抗体诊断试剂盒的研制

Development of Recombinant Antigen of Human T-lymphotropic Virus and Double-antigen Sandwich ELISA

为研制灵敏、特异的抗人类T淋巴细胞白血病病毒(HTLV)抗体诊断试剂,将一段HTLV Ⅰ和HTLV Ⅱenv区嵌合基因在大肠杆菌中表达,获得的重组抗原具有良好活性。将该抗原作为酶标记抗原建立双抗原夹心法ELISA(dsELISA),对31份HTLV Ⅰ型血清和19份HTLV Ⅱ型血清均能100%检出,而在5065份各种阴性血清中特异度为99 94%。用dsELISA试剂与进口间接法试剂(GeneLabs试剂)平行检测18份HTLV Ⅰ参比血清、17份HTLV Ⅱ参比血清和1024份献血员血清,结果dsELISA试剂正确率为100%,对HTLV Ⅰ型血清和HTLV Ⅱ型血清的反应性基本相同,平均s/co值显著高于GeneLabs试剂。而GeneLabs试剂对HTLV Ⅱ型血清的反应性显著弱于Ⅰ型,并有2份BBI参比血清中的Ⅱ型血清漏检。另外,GeneLabs试剂在献血员血清中出现9例假阳性,特异性显著低于dsELISA试剂。这些结果表明:所研制的dsELISA试剂可用于HTLV Ⅰ型和Ⅱ型血清的检测,其灵敏度和特异度均优于进口间接法诊断试剂。

For development of a sensitive and specific diagnostic kit of the human T-lymphotropic virus (HTLV), a chimeric envelope gene of the HTLV-Ⅰ and HTLV-Ⅱ was expressed in E. coli. The recombinant antigen, ENV168 (reacted good with anti-HTLV and was used as peroxidase conjugated antigen), together with another recombinant antigen was developed as a double-antigen sandwich ELISA(dsELISA). The dsELISA could detect out all the 31 HTLV-Ⅰ positive sera and 19 HTLV-Ⅱ positive sera, and the sensitivity of the assay on 5,065 negative sera was 99.94%. A parallel detection of 18 HTLV-Ⅰ positive sera, 17 HTLV-Ⅱ positive sera and 1,024 donors′ sera using dsELISA and a commercial kit (GeneLabs kit) was performed. The results showed that both the sensitivity and specificity of the dsELISA were 100%, and the reactivity (mean s/co value) to HTLV-Ⅰ samples was similar as to HTLV-Ⅱ samples, both were significantly higher than those of GeneLabs kit. For GeneLabs kit, the reactivity with HTLV-Ⅰ samples was significantly higher than that with HTLV-Ⅱ samples. The specificity of dsELISA was significantly higher than GeneLabs kit. These results suggest that the dsELISA kit is useful on the diagnosis of the HTLV-Ⅰ infection as well as HTLV-Ⅱ infection.