Manocha M, Chitralekha K T, Thakar M, Shashikiran D, Paranjape R S, Rao D N
Department of Biochemistry, All India Institute of Medical Sciences, Ansari Nagar, 110029, New Delhi, India.
Immunol Lett. 2003 Feb 3;85(3):275-8. doi: 10.1016/s0165-2478(02)00257-2.
Serological diagnosis of human immunodeficiency virus (HIV) based on detection of HIV antibodies is one of the easiest, cheapest and simplest assay. Synthetic peptides corresponding to immunodominant regions of envelope glycoprotein (gp41, V3 loop for HIV-1 and gp36 for HIV-2) were used in the present study, to detect the anti-HIV antibodies in sera of Sexually Transmitted Diseases (STD), Tuberculosis (TB), Anti-Natal Care (ANC) patients. About 550 serum samples were tested using Enzyme Linked Immunosorbent Assay (ELISA) technique. The human sera positive for antibody to HIV-1 and HIV-2, reacted to different degrees with these peptides when used as a plain peptide with or without CGG motif/biotin motif at the amino terminus. The selected sequences are of Indian strain with 'C' serotype. The results showed a 100% sensitivity and specificity for V3 loop peptide and 98% sensitivity and specificity for gp41 peptide containing CGG moiety while the plain peptides showed similar sensitivities but low specificity's, i.e. 98% for V3 loop peptide and 42% for gp41 peptide when reacted with HIV-1 positive sera. The presence of biotin at the amino terminus did not provide any beneficial effect in increasing the sensitivity although the specificity was enhanced for both the peptide sequences, i.e. gp41 and V3 loop peptide. Furthermore, the gp36 peptide containing CGG moiety detected the HIV-2 sera with 100% sensitivity and 98% specificity while the sensitivity and specificity of gp36 plain peptide was reduced to 98 and 90%. Thus the study overall highlighted the importance of synthetic peptides containing CGG moiety as a capture antigen in detecting both HIV-1 & 2 sera using an indigenously built ELISA system which is simple, cheap, sensitive and cost effective for rural areas.
基于检测人类免疫缺陷病毒(HIV)抗体的血清学诊断是最简单、最便宜且最简便的检测方法之一。本研究使用了与包膜糖蛋白免疫显性区域相对应的合成肽(HIV-1的gp41、V3环以及HIV-2的gp36),以检测性传播疾病(STD)、结核病(TB)、产前保健(ANC)患者血清中的抗HIV抗体。使用酶联免疫吸附测定(ELISA)技术检测了约550份血清样本。当作为在氨基末端带有或不带有CGG基序/生物素基序的普通肽使用时,对HIV-1和HIV-2抗体呈阳性的人血清与这些肽有不同程度的反应。所选序列为具有“C”血清型的印度毒株。结果显示,V3环肽的敏感性和特异性均为100%,含CGG部分的gp41肽的敏感性和特异性分别为98%,而普通肽的敏感性相似但特异性较低,即与HIV-1阳性血清反应时,V3环肽为98%,gp41肽为42%。氨基末端存在生物素虽然增强了两种肽序列(即gp41和V3环肽)的特异性,但在提高敏感性方面没有任何有益效果。此外,含CGG部分的gp36肽检测HIV-2血清的敏感性为100%,特异性为98%,而gp36普通肽的敏感性和特异性分别降至98%和90%。因此,该研究总体突出了含CGG部分的合成肽作为捕获抗原在使用本土构建的ELISA系统检测HIV-1和HIV-2血清中的重要性,该系统简单、便宜、灵敏且对农村地区具有成本效益。