Heneine W, Yamamoto S, Switzer W M, Spira T J, Folks T M
Retrovirus Diseases Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA.
J Infect Dis. 1995 May;171(5):1210-6. doi: 10.1093/infdis/171.5.1210.
In an ultrasensitive assay for reverse transcriptase (RT), an in vitro-transcribed heteropolymeric RNA sequence was used as a template and polymerase chain reaction (PCR) amplification with Southern blot hybridization served as a detection system for the cDNA reaction product. The assay, called Amp-RT, detected 9 tested retroviruses in unconcentrated culture supernatants diluted 10(2)- to 10(5)-fold. A comparative analysis using human immunodeficiency virus type 1 (HIV-1) revealed that Amp-RT was 100,000 times more sensitive than the standard RT assay, 10,000 times more sensitive than p24 antigen capture and branched DNA assays, and 100 times more sensitive than RT-PCR or TCID50 assays. Analysis of serum specimens from 42 HIV-1-infected persons by Amp-RT showed that 36 samples (85.7%) were RT-positive. In contrast, 41 serum specimens from persons seronegative for HIV-1 and human T lymphotropic virus types I and II were all Amp-RT-negative.
在一种用于逆转录酶（RT）的超灵敏检测方法中，体外转录的异聚RNA序列被用作模板，以聚合酶链反应（PCR）扩增结合Southern印迹杂交作为cDNA反应产物的检测系统。这种名为Amp-RT的检测方法，能在稀释10²至10⁵倍的未浓缩培养上清液中检测出9种受试逆转录病毒。使用1型人类免疫缺陷病毒（HIV-1）进行的对比分析表明，Amp-RT的灵敏度比标准RT检测方法高100,000倍, 比p24抗原捕获和分支DNA检测方法高10,000倍，比RT-PCR或TCID50检测方法高100倍。通过Amp-RT对42名HIV-1感染者的血清样本进行分析，结果显示36份样本（85.7%）RT呈阳性。相比之下，来自HIV-1以及I型和II型人类嗜T细胞病毒血清学阴性者的41份血清样本Amp-RT均为阴性。
