用于测定1型人类免疫缺陷病毒RNA的自动样本制备及定量聚合酶链反应LCx检测法的评估
Evaluation of automated sample preparation and quantitative PCR LCx assay for determination of human immunodeficiency virus type 1 RNA.
作者信息
Muller Zsofia, Stelzl Evelyn, Bozic Michael, Haas Josef, Marth Egon, Kessler Harald H
机构信息
Microbiological Laboratory, Regional Public Health Center, H-8000 Szekesfehervar, Hungary.
出版信息
J Clin Microbiol. 2004 Apr;42(4):1439-43. doi: 10.1128/JCM.42.4.1439-1443.2004.
Efforts have been made to achieve full automation of molecular assays for quantitative detection of human immunodeficiency virus type 1 (HIV-1). In the present study, the Abbott LCx HIV RNA Quantitative assay was evaluated in conjunction with automated HIV-1 RNA extraction on the MagNA Pure LC instrument and compared to the conventional LCx HIV RNA Quantitative assay, which uses a manual nucleic acid extraction protocol. Accuracy, linearity, and interassay and intra-assay variations were determined. The performance of the assay in a routine clinical laboratory was tested with a total of 105 clinical specimens. When the accuracy of the LCx HIV RNA Quantitative assay with the automated sample preparation protocol was tested, all results were found to be within +/- 0.5 log unit of the expected results. Determination of linearity resulted in a quasilinear curve over 3.5 log units. For determination of interassay variation, coefficients of variation were found to be between 21 and 66% for the LCx HIV RNA Quantitative assay with the automated sample preparation protocol and between 10 and 69% for the LCx HIV RNA Quantitative assay with manual sample preparation. For determination of intra-assay variation, coefficients of variation were found to be between 7 and 25% for the LCx HIV RNA Quantitative assay with the automated sample preparation protocol and between 7 and 19% for the LCx HIV RNA Quantitative assay with manual sample preparation. When clinical samples were tested by the LCx HIV RNA Quantitative assay with the automated sample preparation protocol and the results were compared with those of the LCx HIV RNA Quantitative assay with manual sample preparation, 95% of all positive results were found to be within +/- 0.5 log unit. In conclusion, the assay with automated sample preparation proved to be suitable for use in the routine diagnostic laboratory and required significantly less hands-on time.
人们一直在努力实现用于定量检测1型人类免疫缺陷病毒（HIV-1）的分子检测的完全自动化。在本研究中，对雅培LCx HIV RNA定量检测法结合MagNA Pure LC仪器上的HIV-1 RNA自动提取进行了评估，并与使用手动核酸提取方案的传统LCx HIV RNA定量检测法进行了比较。测定了准确性、线性以及批间和批内变异。使用总共105份临床标本对该检测法在常规临床实验室中的性能进行了测试。当测试采用自动样品制备方案的LCx HIV RNA定量检测法的准确性时，发现所有结果均在预期结果的±0.5对数单位范围内。线性测定得出了超过3.5对数单位的准线性曲线。对于批间变异的测定，采用自动样品制备方案的LCx HIV RNA定量检测法的变异系数在21%至66%之间，采用手动样品制备的LCx HIV RNA定量检测法的变异系数在10%至69%之间。对于批内变异的测定，采用自动样品制备方案的LCx HIV RNA定量检测法的变异系数在7%至25%之间，采用手动样品制备的LCx HIV RNA定量检测法的变异系数在7%至19%之间。当采用自动样品制备方案的LCx HIV RNA定量检测法对临床样品进行检测并将结果与采用手动样品制备的LCx HIV RNA定量检测法的结果进行比较时，发现所有阳性结果的95%在±0.5对数单位范围内。总之，采用自动样品制备的检测法被证明适用于常规诊断实验室，且所需的实际操作时间显著减少。
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