Detection of HIV-1 distribution in different blood fractions by two nucleic acid amplification assays.
Detection of HIV-1 distribution in different blood fractions by two nucleic acid amplification assays.
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通过两种核酸扩增测定检测不同血液成分中 HIV-1 的分布。
DOI：
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发表时间：
1993
影响因子：
1.5
通讯作者：
H. Huisman
中科院分区：
文献类型：
--
作者：
S. Bruisten;B. van Gemen;M. Koppelman;M. Rasch;D. van Strijp;R. Schukkink;R. Beyer;H. Weigel;P. Lens;H. Huisman
一种新的扩增方法——核酸序列依赖性扩增（NASBA）与聚合酶链反应（PCR）一起被用于检测感染艾滋病毒和未感染样本的不同血液成分中的艾滋病毒1型（HIV - 1）序列。我们检测了全血、血浆、外周血单个核细胞（PBMC）和血小板。通过PCR、逆转录酶 - PCR（RT - PCR）或NASBA，在37名未感染者的血液成分中均未发现HIV - 1序列。我们发现感染的血浆样本中均不含有HIV - 1 DNA序列。然而，这些血浆样本中有很高比例的HIV - 1 RNA呈阳性：NASBA检测为86%，RT - PCR检测为80%。NASBA和RT - PCR在逐个样本基础上的一致性为91%。只有33%的血浆样本HIV - 1 p24抗原呈阳性，这表明扩增方法具有更高的敏感性。我们发现几乎所有的PBMC成分中的HIV - 1（前）病毒序列都呈阳性（99%的HIV - 1 DNA阳性，91%的HIV - 1 RNA阳性）。大量的血小板成分含有HIV - 1 RNA，这通过RT - PCR和NASBA的阳性结果得以证明。我们发现一方面CD4⁺T细胞计数和T细胞反应性与另一方面在所有血液成分中通过PCR、RT - PCR和NASBA检测到HIV - 1序列之间呈反比关系。对PBMC中HIV - 1 PCR信号的定量显示前病毒滴度与CD4⁺水平呈反比关系。这一发现支持了早期的观察结果，即临床疾病和低CD4⁺细胞计数与病毒载量增加有关。
A new amplification procedure, NASBA (nucleic acid sequence-based amplification), was used together with the polymerase chain reaction (PCR) to detect HIV-1 sequences in different blood fractions of both HIV-infected and uninfected samples. We tested whole blood, plasma, peripheral blood mononuclear cells (PBMCs), and platelets. No HIV-1 sequences were found in blood fractions of 37 uninfected persons either by PCR, reverse transcriptase-PCR (RT-PCR), or NASBA. We found that none of the infected plasma samples contained HIV-1 DNA sequences. However, a high percentage of these plasma samples was positive for HIV-1 RNA: 86% by NASBA and 80% by RT-PCR. The concordance on a sample-to-sample basis of NASBA and RT-PCR was 91%. Only 33% of the plasma samples was HIV-1 p24-antigen positive, demonstrating the superior sensitivity of amplification procedures. We found that almost all PBMC fractions were positive for HIV-1 (pro)viral sequences (99% HIV-1 DNA positive, 91% HIV-1 RNA positive). A large proportion of the platelet fractions contained HIV-1 RNA, as demonstrated by positive RT-PCR and NASBA results. We found an inverse relation between CD4+ T cell count and T cell reactivity on the one hand and detection of HIV-1 sequences by PCR, RT-PCR, and NASBA on the other hand in all blood fractions. Quantification of the HIV-1 PCR signal in PBMCs revealed an inverse relation of proviral titers with CD4+ levels. This finding supports earlier observations that clinical disease and low CD4+ cell counts are related to an increased viral burden.
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影响因子：
39.2
作者：
SCHNITTMAN, SM;GREENHOUSE, JJ;LANE, HC
通讯作者：
LANE, HC
DOI：
10.1073/pnas.86.4.1173
发表时间：
1989-02-01
影响因子：
11.1
作者：
KWOH, DY;DAVIS, GR;GINGERAS, TR
通讯作者：
GINGERAS, TR
影响因子：
158.5
作者：
HO, DD;MOUDGIL, T;ALAM, M
通讯作者：
ALAM, M
DOI：
10.1073/pnas.87.5.1874
发表时间：
1990-03-01
影响因子：
11.1
作者：
GUATELLI, JC;WHITFIELD, KM;GINGERAS, TR
通讯作者：
GINGERAS, TR