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通过竞争性聚合酶链反应对意大利注射吸毒者、中非人和美洲印第安人的外周血单个核细胞中的HTLV-II前病毒拷贝进行定量分析。

Quantification of HTLV-II proviral copies by competitive polymerase chain reaction in peripheral blood mononuclear cells of Italian injecting drug users, central Africans, and Amerindians.

作者信息

Cimarelli A, Duclos C A, Gessain A, Cattaneo E, Casoli C, Biglione M, Mauclère P, Bertazzoni U

机构信息

Istituto di Genetica Biochimica ed Evoluzionistica del CNR, Pavia, Italy.

出版信息

J Acquir Immune Defic Syndr Hum Retrovirol. 1995 Oct 1;10(2):198-204. doi: 10.1097/00042560-199510020-00014.

Abstract

To better correlate the burden of human T cell leukemia virus type I (HTLV-I) and type II (HTLV-II) infection with diagnostic and prognostic markers, we developed a new competitive polymerase chain reaction (PCR) assay for the quantitative determination of proviral copy numbers in infected cells. A competitive plasmid was constructed that carried a 112-bp fragment from a highly conserved region of the HTLV tax gene and that was further modified by inserting a sequence of 24 bp. This competitive PCR assay system can be used for the quantification of HTLV-I and HTLV-II proviral DNA as demonstrated by using HTLV-I- and HTLV-II-infected cell lines and/or patient material. We determined the HTLV-II proviral load in peripheral blood mononuclear cells (PBMCs) of 11 Italian injecting drug users (IDUs) infected by this virus and in PBMCs of 10 seropositive Amerindian and Central African individuals from endemically infected ethnic groups. A great variation was observed in the number of HTLV-II proviral sequences in the PBMCs of Italian drug abusers, ranging from 5-10 to 16,239 copies/10(5) cells. There was no clear-cut correlation between proviral load, CD8 count, stage of HIV-1 infection, and therapy. A considerable variation in HTLV-II proviral load was also observed in PBMCs of Amerindians and Central Africans with no correlation between the amount of HTLV-II provirus and the geographic origin of the infected individuals.

摘要

为了更好地将人类I型嗜T细胞病毒(HTLV-I)和II型嗜T细胞病毒(HTLV-II)感染的负担与诊断和预后标志物相关联,我们开发了一种新的竞争性聚合酶链反应(PCR)检测方法,用于定量测定感染细胞中的前病毒拷贝数。构建了一个竞争性质粒,其携带来自HTLV tax基因高度保守区域的112 bp片段,并通过插入一个24 bp的序列进行了进一步修饰。如使用HTLV-I和HTLV-II感染的细胞系和/或患者材料所证明的,这种竞争性PCR检测系统可用于HTLV-I和HTLV-II前病毒DNA的定量。我们测定了11名感染该病毒的意大利注射吸毒者(IDU)外周血单个核细胞(PBMC)中的HTLV-II前病毒载量,以及10名来自地方性感染族群的血清阳性美洲印第安人和中非个体的PBMC中的HTLV-II前病毒载量。在意大利吸毒者的PBMC中观察到HTLV-II前病毒序列数量有很大差异,范围从5 - 10到16,239拷贝/10(5)细胞。前病毒载量、CD8计数、HIV-1感染阶段和治疗之间没有明确的相关性。在美洲印第安人和中非人的PBMC中也观察到HTLV-II前病毒载量有相当大的差异,HTLV-II前病毒数量与受感染个体的地理来源之间没有相关性。

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