Instituto Nacional de Tecnología Agropecuaria (INTA), CICVyA, Instituto de Virología, Buenos Aires, Argentina
Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Buenos Aires, Argentina.
J Clin Microbiol. 2018 Jun 25;56(7). doi: 10.1128/JCM.00304-18. Print 2018 Jul.
Quantitative real-time PCR (qPCR) is increasingly being used for the detection of bovine leukemia virus (BLV) proviral DNA. Nevertheless, quality control for the validation and standardization of such tests is currently lacking. Therefore, the present study was initiated by three Office International des Epizooties (OIE) reference laboratories and three collaborating laboratories to measure the interlaboratory variability of six already developed and available BLV qPCR assays. For that purpose, an international panel of 58 DNA samples reflecting the dynamic range of the majority of the assays was distributed to six testing centers. Based on qualitative results, the overall agreement among all six laboratories was moderate. However, significant variability in the measurement of the BLV proviral DNA copy number was observed among different laboratories. Quantitative PCR assays, even when performed by experienced staff, can yield large variability in BLV proviral DNA copy numbers without harmonization. Further standardization of different factors (i.e., utilization of unified protocols and unique calibrators) should increase interlaboratory agreement.
定量实时 PCR（qPCR）越来越多地用于检测牛白血病病毒（BLV）前病毒 DNA。然而，目前缺乏此类检测的验证和标准化的质量控制。因此，本研究由三个国际动物卫生组织（OIE）参考实验室和三个合作实验室发起，旨在测量六个已开发和可用的 BLV qPCR 检测方法的实验室间变异性。为此，一个反映大多数检测方法动态范围的 58 个 DNA 样本国际小组分发给六个测试中心。基于定性结果，所有六个实验室之间的总体一致性为中等。然而，在不同实验室之间观察到 BLV 前病毒 DNA 拷贝数的测量存在显著差异。定量 PCR 检测方法，即使由经验丰富的人员进行操作，也可能在没有协调的情况下导致 BLV 前病毒 DNA 拷贝数的大量变异性。进一步标准化不同因素（即，利用统一的协议和独特的校准品）应提高实验室间的一致性。
