HIV核心抗原p24检测方法的建立及应用

Establishment and application of a sandwich ELISA assay for HIV p24

ＨＩＶ感染的检测在预防ＡＩＤＳ传播方面有重要作用。本研究以重组抗原ｐＧ１免疫Ｂａｌｂ／ｃ小鼠，应用淋巴细胞杂交瘤技术建立了５株识别ＨＩＶ核心抗原ｐ２４的杂交瘤细胞株Ｄ３，１Ｈ１，２Ｇ１０，３Ｈ５和４Ｈ６。经鉴定这５株ＭｃＡｂ与其它病毒抗原无交叉反应，分别识别３个不同的抗原表位。应用两株识别不同表位的ＭｃＡｂ２Ｇ１０和３Ｈ５建立了检测ＨＩＶｐ２４抗原的夹心ＥＬＩＳＡ法，并初步检测了２４份ＨＩＶ抗体阳性血清。

Recombinant protein pG1 containing total sequence of HIV core antigen p24 was used as an immunogen to immunize Balb/c mice and five hybridomas secreting McAbs to p24, D3, 1H1, 2G10, 3H5 and 4H6, were raised by conventional B lymphocyte hybridoma technique. The Ig subclass, recognized epitope and specificity of obtained McAbs were identified. The results showed that these five McAbs recognized three different epitopes of p24 molecule respectively and did not crossreact with HBcAg, HCV C region and NS3 region and E.coli protein. Using two McAbs recognizing two distinct epitopes, a sandwich ELISA method for measurement of HIV p24 was established with sensitivity about 1 ng/ml.