Schutten M, van den Hoogen B, van der Ende M E, Gruters R A, Osterhaus A D, Niesters H G
Department of Virology, Erasmus Medical Centre Rotterdam, The Netherlands.
J Virol Methods. 2000 Jul;88(1):81-7. doi: 10.1016/s0166-0934(00)00177-4.
An assay is described for the quantification of human immunodeficiency virus type 2 (HIV-2) RNA in EDTA plasma based on RT-PCR using the Taqman real-time PCR detection method. As standard, an electron microscopically counted virus stock of HIV-2 strain NIHZ was used. The lower detection limit is 5 # 102 HIV-2 RNA copies per ml of EDTA plasma. The assay is linear within the range required (5 # 102-106 HIV-2 RNA copies/ml of EDTA plasma) with an intra assay variability of 2.5% and an inter-assay variability ranging from 2% at 106 copies to 7.5% at the lower detection limit. Three primer/probe combinations were developed to circumvent false negative samples due to nucleotide variation in the target sequence. Using these primer/probe sets enabled the detection of HIV-2 DNA sequences from all HIV-2 seropositive individuals and two out of five dual human immunodeficiency virus type 1 (HIV-1) and HIV-2 seropositive individuals visiting the University Hospital Rotterdam.
描述了一种基于Taqman实时PCR检测方法的逆转录-聚合酶链反应（RT-PCR），用于定量测定乙二胺四乙酸（EDTA）血浆中的2型人类免疫缺陷病毒（HIV-2）RNA。作为标准品，使用了经电子显微镜计数的HIV-2 NIHZ毒株病毒储备液。检测下限为每毫升EDTA血浆5×10²个HIV-2 RNA拷贝。该检测方法在所需范围内（每毫升EDTA血浆5×10² - 10⁶个HIV-2 RNA拷贝）呈线性，批内变异系数为2.5%，批间变异系数在10⁶个拷贝时为2%，在检测下限处为7.5%。开发了三种引物/探针组合，以规避由于靶序列核苷酸变异导致的假阴性样本。使用这些引物/探针组能够检测所有HIV-2血清学阳性个体以及鹿特丹大学医院就诊的五名1型和2型人类免疫缺陷病毒（HIV-1和HIV-2）双重血清学阳性个体中的两名的HIV-2 DNA序列。
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