Detection of human immunodeficiency virus type 1 reverse transcriptase using aptamers as probes in affinity capillary electrophoresis

被引：88

作者：

Pavski, V ^{[1
]}

Le, XC ^{[1
]}

机构：

[1] Univ Alberta, Dept Publ Hlth Sci, Environm Hlth Sci Program, Edmonton, AB T6G 2G3, Canada

来源：

ANALYTICAL CHEMISTRY | 2001年 / 73卷 / 24期

关键词：

D O I：

10.1021/ac0107305

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

An affinity capillary electrophoresis/laser-induced fluorescence (CE/LIF) assay was developed for direct and specific detection of reverse transcriptase (RI) of the type I human immunodeficiency virus (HIV-1) using fluorescently labeled single-stranded DNA aptamers as probes. The aptamer used (Rr 26) is specific for HIV-1 RT, and it exhibited no cross-reactivity with RTs of the enhanced avian myeloblastosis virus (AMV), the Moloney murine leukemia virus (MMLV), or denatured HIV-1 RT An affinity complex of Rr 26-HIV-1 RT was readily formed, and calibration curves were linear up to 50 nM (6 mug/mL) HIV-1 RT concentration, with both the free probe and complex peak usable for analytical quantitation. Cell culture media (RPMI with 10% fetal bovine serum) interfered with the assay and aptamer-HIV-1 RT binding. Nonspecific binding was observed in low or undiluted culture, necessitating at least 100-fold dilution for analysis of raw culture samples.

引用

页码：6070 / 6076

页数：7

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