Liu Tao, Liu Ji Feng, Yu Hua, Si Guo Jing, Hu Jun, Li Jun
Microbiology Laboratory, Hangzhou Center for Disease Control and Prevention, Hangzhou, People's Republic of China.
Department of Dermatology, The Third People's Hospital of Hangzhou, Hangzhou, People's Republic of China.
Singapore Med J. 2015 Jun;56(6):346-52. doi: 10.11622/smedj.2014197.
Herpes simplex virus type 2 (HSV-2) is the most common cause of genital herpes. Glycoprotein G (gG) is a prototype antigen for type-specific serodiagnosis distinguishing between HSV type 1 (HSV-1) and HSV-2 infections. As immunological diagnosis kits for accurate differentiation between HSV-1 and HSV-2 antibodies can be expensive, there is a need to develop a convenient, sensitive, specific and cost-effective serodiagnostic kit.
We successfully expressed a fragment of gG comprising residues 321-580 of HSV-2 with histidine tag (gG(321-580His)) in a Bac-to-Bac baculovirus expression system, which had an antigenicity similar to its native counterpart. An indirect enzyme-linked immunosorbent assay (ELISA) was developed using gG(321-580His) as the diagnostic antigen and evaluated by comparison with a commercial HerpeSelect 2 ELISA immunoglobulin G kit as reference.
In testing 318 field serum samples, the diagnostic relative sensitivity and specificity of the developed gG(321-580His)-ELISA test in qualitative comparison with the commercial kit were 93.81% and 96.74%, respectively, and the accuracy was 94.65%.
The study indicates that gG(321-580His) has a high diagnostic potential for HSV-2 virus serodiagnosis in humans.
单纯疱疹病毒2型(HSV-2)是生殖器疱疹最常见的病因。糖蛋白G(gG)是用于区分1型单纯疱疹病毒(HSV-1)和HSV-2感染的型特异性血清学诊断的原型抗原。由于用于准确区分HSV-1和HSV-2抗体的免疫诊断试剂盒可能价格昂贵,因此需要开发一种方便、灵敏、特异且经济高效的血清学诊断试剂盒。
我们在杆状病毒表达系统中成功表达了包含HSV-2第321-580位残基且带有组氨酸标签的gG片段(gG(321-580His)),其抗原性与其天然对应物相似。使用gG(321-580His)作为诊断抗原开发了一种间接酶联免疫吸附测定(ELISA),并与商用HerpeSelect 2 ELISA免疫球蛋白G试剂盒作为参考进行比较评估。
在检测318份现场血清样本时,与商用试剂盒进行定性比较,所开发的gG(321-580His)-ELISA检测的诊断相对敏感性和特异性分别为93.81%和96.74%,准确性为94.65%。
该研究表明gG(321-580His)在人类HSV-2病毒血清学诊断中具有很高的诊断潜力。