Evaluation of the Roche COBAS^® TaqMan^® HIV-1 test for quantifying HIV-1 RNA in infected cells and lymphoid tissue

详细信息    查看全文

• 作者：Cristina Gil^a ; ^{cgil@clinic.ub.es"" rel=""nofollow} ; M. Teresa Garcia^a ; Felipe Garcia^a ; ^b ; Jos&#xe9 ; M. Miró ; ^a ; ^b ; Fernando Agü ; ero^a ; ¹ ; Luc&#xed ; a Aló ; s^d ; Laura Zamora^a ; Alicia Capó ; n^c ; Josep Costa^c ; Tom&#xe1 ; s Pumarola^c ; Jos&#xe9 ; M. Gatell^a ; ^b
• 关键词：HIV-1 RNA quantitation ; Lymphoid tissue ; COBAS TaqMan HIV-1
• 刊名：Journal of Virological Methods
• 出版年：2011
• 出版时间：June 2011
• 年：2011
• 卷：174
• 期：1-2
• 页码：69-76
• 全文大小：285 K

文摘

Lymphoid tissue is the main reservoir of HIV-1 in infected individuals. In this study the COBAS^® TaqMan^® HIV-1 test was evaluated for use with the High Pure System (HPS), for quantifying HIV-1 RNA in infected cells and lymphoid tissue specimens. Serial dilutions of 8E5–LAV1 infected T-cells into SUP-T1 cells and 44 tonsil specimens were examined. Some modifications of the test were required, such as the removal of residual DNA and the HIV-1 RNA output copies were adjusted to the sample input and expressed as HIV-1 RNA copies/μg of total RNA. The Roche COBAS^® TaqMan HIV-1^® (HPS) test proved to be a robust, sensitive, specific and reproducible method for quantifying HIV-1 RNA in infected cells and lymphoid tissue. Linearity and reproducibility were observed in serial dilutions of 8E5–LAV1 infected T-cells (R² > 0.86). High reproducibility was found in clinical tonsil specimens (Wilcoxon test p > 0.05). rDNase I treatment was essential to avoid false positives caused by residual HIV-1 DNA, mainly in tonsil specimens obtained from infected patients receiving effective antiretroviral treatment. Probit analysis determined the limit of detection as 22 HIV-1 RNA copies/μg of total RNA. The Roche COBAS^® TaqMan^® HIV-1 (HPS) test thereby proved to be a helpful tool for measuring the HIV-1 viral load in infected cells and lymphoid tissue reservoirs.