Jurriaans S, Dekker J T, de Ronde A
Human Retrovirus Laboratory, University of Amsterdam, The Netherlands.
AIDS. 1992 Jul;6(7):635-41. doi: 10.1097/00002030-199207000-00004.
To determine viral DNA load in peripheral blood mononuclear cells (PBMC) from HIV-1-infected individuals.
HIV-1 copy numbers were determined using a quantitative polymerase chain reaction (PCR), the PCR-aided template titration assay (PATTY). PATTY utilizes an internal plasmid control DNA, which is amplified within the same tube and using the same primers as the PBMC target DNA. HIV-1 copy numbers were confirmed by limiting-dilution PCR analysis.
PBMC viral load of 19 long-term (greater than 4 years) HIV-1-infected individuals ranged from 0.8 to 100 copies per 10(3) PBMC. Significantly higher copy numbers were found among p24-antigen-positive than among p24-antigen-negative individuals. In addition, the PBMC viral load of two HIV-1-infected individuals was monitored during the first 3 months after acute infection. For both patients, the HIV-1 copy numbers were shown to peak at the time of HIV-1-antibody seroconversion and decline subsequently (range, 0.6-10 copies per 10(3) PBMC).
PATTY is a useful method for assessing the HIV-1 copy numbers in PBMC DNA. Viral DNA load peaks shortly after infection and reaches an individual specific level that is probably stable within a few months of infection. Viral DNA load in PBMC varies widely among long-term HIV-1-infected individuals.
测定HIV-1感染个体外周血单个核细胞(PBMC)中的病毒DNA载量。
采用定量聚合酶链反应(PCR)即PCR辅助模板滴定法(PATTY)测定HIV-1拷贝数。PATTY利用内部质粒对照DNA,其在同一管内与PBMC靶DNA使用相同引物进行扩增。通过有限稀释PCR分析确认HIV-1拷贝数。
19名长期(超过4年)HIV-1感染个体的PBMC病毒载量为每10³个PBMC中0.8至100拷贝。p24抗原阳性个体的拷贝数显著高于p24抗原阴性个体。此外,对两名HIV-1感染个体在急性感染后的前3个月进行了PBMC病毒载量监测。两名患者的HIV-1拷贝数均在HIV-1抗体血清转化时达到峰值,随后下降(范围为每10³个PBMC中0.6至10拷贝)。
PATTY是评估PBMC DNA中HIV-1拷贝数的有用方法。病毒DNA载量在感染后不久达到峰值,并在感染后几个月内达到个体特异性水平且可能保持稳定。长期HIV-1感染个体的PBMC中病毒DNA载量差异很大。