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核酸序列依赖性扩增(NASBA)HIV-1 RNA扩增系统针对不同HIV-1亚型的反应性和扩增效率。

Reactivity and amplification efficiency of the NASBA HIV-1 RNA amplification system with regard to different HIV-1 subtypes.

作者信息

Gobbers E, Fransen K, Oosterlaken T, Janssens W, Heyndrickx L, Ivens T, Vereecken K, Schoones R, van de Wiel P, van der Groen G

机构信息

Organon Teknika, Turnhout, Belgium.

出版信息

J Virol Methods. 1997 Jul;66(2):293-301. doi: 10.1016/s0166-0934(97)00072-4.

Abstract

In view of the genetic diversity of the human immunodeficiency virus Type 1, we assessed the sensitivity and quantification efficiency of the HIV-1 RNA NASBA amplification system with respect to different HIV-1 subtypes and recombinants. Twenty cell culture supernatants representing 17 HIV-1 group M and 3 group O strains were tested, and NASBA RNA loads were compared with results obtained with a RT-PCR based HIV-1 RNA quantitation method, with p24-antigen concentrations and with the infective dose. The current HIV-1 RNA NASBA seemed suitable to quantitate representatives of different HIV-1 M subtypes. Differences between NASBA and RT-PCR loads were observed for certain HIV-1 M strains. Significantly lower RT-PCR loads were measured for most gag A, gag B and gag F strains, whereas NASBA detected lower copy numbers in 1 gag H strain and 1 gag H/env G recombinant. NASBA was not able to quantify 1 HIV-1 group M recombinant. Some of these differences could be explained by the presence and position of mismatches with primers. HIV-1 group O strains were not detectable by both RNA amplification methods. A firm correlation was not observed between the measured RNA loads and either the p24-antigen concentration or the infective dose.

摘要

鉴于人类免疫缺陷病毒1型(HIV-1)的基因多样性,我们评估了HIV-1 RNA核酸序列扩增技术(NASBA)扩增系统针对不同HIV-1亚型和重组体的敏感性及定量效率。对代表17种HIV-1 M组和3种O组毒株的20份细胞培养上清液进行了检测,并将NASBA RNA载量与基于逆转录聚合酶链反应(RT-PCR)的HIV-1 RNA定量方法所得结果、p24抗原浓度以及感染剂量进行了比较。当前的HIV-1 RNA NASBA似乎适合对不同HIV-1 M亚型的代表毒株进行定量。对于某些HIV-1 M毒株,观察到了NASBA与RT-PCR载量之间的差异。对于大多数gag A、gag B和gag F毒株,RT-PCR检测到的载量显著更低,而NASBA在1株gag H毒株和1株gag H/env G重组体中检测到较低的拷贝数。NASBA无法对1株HIV-1 M组重组体进行定量。其中一些差异可以通过与引物错配的存在及位置来解释。两种RNA扩增方法均无法检测到HIV-1 O组毒株。在所测RNA载量与p24抗原浓度或感染剂量之间未观察到确定的相关性。

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