Wang J, Cai X, Rivas G, Shiraishi H, Farias P A, Dontha N
Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces 88003, USA.
Anal Chem. 1996 Aug 1;68(15):2629-34. doi: 10.1021/ac9602433.
An electrochemical biosensor for the detection of short DNA sequences related to the human immunodeficiency virus type 1 (HIV-1) is described. The sensor relies on the immobilization and hybridization of the 21- or 42-mer single-stranded oligonucleotide from the HIV-1 U5 long terminal repeat (LTR) sequence at carbon paste or strip electrodes. The extent of hybridization between the complementary sequences is determined by the enhancement of the chronopotentiometric peak of the Co(phen)3(3+) indicator. Numerous factors affecting the probe immobilization, target hybridization, and indicator binding reactions are optimized to maximize the sensitivity and speed the assay time. A detection limit of 4 x 10(-9) M HIV-1 U5 LTR segment is reported following a 30 min hybridization. The hybridization biosensor format obviates the use of radioisotopes common in radioactive methods for the detection of HIV-1 DNA. We also report on the direct adsorptive chronopotentiometric stripping measurements of trace levels of various HIV-1 DNAs.
本文描述了一种用于检测与人类免疫缺陷病毒1型（HIV-1）相关的短DNA序列的电化学生物传感器。该传感器依赖于来自HIV-1 U5长末端重复序列（LTR）的21或42聚体单链寡核苷酸在碳糊或条形电极上的固定和杂交。互补序列之间的杂交程度由Co(phen)3(3+)指示剂的计时电位峰增强来确定。对影响探针固定、靶标杂交和指示剂结合反应的众多因素进行了优化，以最大限度地提高灵敏度并缩短检测时间。在30分钟杂交后，报告的HIV-1 U5 LTR片段检测限为4×10(-9) M。这种杂交生物传感器形式避免了在检测HIV-1 DNA的放射性方法中常用的放射性同位素的使用。我们还报告了各种HIV-1 DNA痕量水平的直接吸附计时电位溶出测量。
