虽然 HC2 和 GP5 /6 PCR-EIA 在新 HPV 检测的测试验证中至关重要，但它们代表了基于不再广泛使用的技术的第一代比较测试。在当前的指南中，提出了第二代比较测试的标准，其中包括更详细的 HPV 基因型解析。第二代比较试验应优先仅针对被归类为致癌的 12 种基因型（IARC-I 组），并且与第一代比较试验相比，对 CIN2 和 CIN3 表现出一致的非劣效敏感性，对 ≤CIN1 具有特异性。至少三项验证研究使用 0.95 的相对敏感性基准和 0.98 的相对特异性基准。验证应考虑使用的存储介质和其他样品处理程序。进行荟萃分析以确定满足这些严格标准的检测方法。四项检测符合新标准：(1) 实时高风险 HPV 检测 (Abbott)、(2) Cobas-4800 HPV 检测（罗氏分子系统）、(3) Onclarity HPV 检测 (BD Diagnostics) 和 (4) Anyplex II HPV HR 检测 (Seegene)，每项均在三到六项研究中进行评估。虽然四种检测针对 14 种致癌基因型，但前两种检测分别识别 HPV16 和 18，第三种检测分别识别五种类型，第四种检测分别识别所有类型。© 2024 作者。 《医学病毒学杂志》由 Wiley periodicals LLC 出版。

While HC2 and GP5+/6+ PCR-EIA were pivotal in test validation of new HPV assays, they represent the first generation of comparator tests based upon technologies that are not in widespread use anymore. In the current guideline, criteria for second-generation comparator tests are presented that include more detailed resolution of HPV genotypes. Second-generation comparator tests should preferentially target only the 12 genotypes classified as carcinogenic (IARC-group I), and show consistent non-inferior sensitivity for CIN2+ and CIN3+ and specificity for ≤CIN1 compared to one of the first-generations comparators, in at least three validation studies using benchmarks of 0.95 for relative sensitivity and 0.98 for relative specificity. Validation should take into account used storage media and other sample handling procedures. Meta-analyses were conducted to identify the assays that fulfill these stringent criteria. Four tests fulfilled the new criteria: (1) RealTime High-Risk HPV Test (Abbott), (2) Cobas-4800 HPV test (Roche Molecular System), (3) Onclarity HPV Assay (BD Diagnostics), and (4) Anyplex II HPV HR Detection (Seegene), each evaluated in three to six studies. Whereas the four assays target 14 carcinogenic genotypes, the first two identify separately HPV16 and 18, the third assay identifies five types separately and the fourth identifies all the types separately.© 2024 The Author(s). Journal of Medical Virology published by Wiley Periodicals LLC.