AMBIOR, Laboratory for Cell Biology and Histology, University of Antwerp, Antwerp, Belgium.
Laboratory of Molecular Pathology, AML, Antwerp, Belgium.
J Med Virol. 2021 Jun;93(6):3841-3848. doi: 10.1002/jmv.26623. Epub 2021 Mar 1.
Cutaneous warts are infectious disorders caused by human papillomavirus (HPV). A recent study revealed that the HPV genotype influences the natural course and response to treatment for plantar warts, suggesting that HPV genotyping could potentially be used to optimize wart treatment schemes. For this purpose, a wart-associated HPV genotyping assay was developed. The assay was subjected to an intensive validation process including, i.a., empiric determination of the annealing temperature, primer-probe optimization, evaluation of the analytical specificity and sensitivity, viral load quantification, and qualitative as well as quantitative analysis of intra-run repeatability and inter-run reproducibility. The newly developed assay was employed in a small-scale HPV genotyping study of wart biopsies (n = 50). The assay exhibited an analytical type-specific sensitivity and specificity of 100% (95% confidence interval [CI]: 83.9%-100%). The limit of quantification of the tested sequences corresponded to less than 17 viral copies/µl, while the limit of detection was less than 5 copies/µl. Very good to excellent agreements were gained between intra- and inter-run measurements (κ = 0.85-1.00) and coefficients of variation of the quantitative agreements were less then 3%. 22.5% (95% CI: 11%-39%) of the analyzed biopsies were negative for the tested HPV types, while 35% (95% CI: 21%-52%) contained multiple infections. The wart-associated HPV quantitative polymerase chain reaction assay was proven to be highly sensitive and specific. Multiple HPV infections were detected in 35% of lesions, contradicting the current literature claiming that in immunocompetent patients only 4%-16% of warts exhibit multiple HPV infections. This assay is qualified to be implemented in development of future genotype specific wart treatment strategies.
皮肤疣是由人乳头瘤病毒（HPV）引起的传染性疾病。最近的一项研究表明，HPV 基因型会影响足底疣的自然病程和治疗反应，这表明 HPV 基因分型可能用于优化疣治疗方案。为此，开发了一种与疣相关的 HPV 基因分型检测方法。该方法经过了严格的验证，包括确定退火温度、优化引物探针、评估分析特异性和灵敏度、病毒载量定量以及分析内运行重复性和外运行重现性的定性和定量分析。新开发的检测方法应用于对 50 例疣活检样本进行 HPV 基因分型的小型研究。该检测方法表现出 100%（95%置信区间 [CI]：83.9%-100%）的分析型特异性灵敏度和特异性。所检测序列的定量下限低于 17 个病毒拷贝/µl，检测下限低于 5 个拷贝/µl。内运行和外运行测量之间的一致性非常好（κ=0.85-1.00），定量一致性的变异系数小于 3%。22.5%（95% CI：11%-39%）的分析活检标本对所检测的 HPV 类型呈阴性，而 35%（95% CI：21%-52%）包含多重感染。与疣相关的 HPV 定量聚合酶链反应检测方法被证明具有高度的灵敏度和特异性。在 35%的病变中检测到多重 HPV 感染，这与目前的文献声称在免疫功能正常的患者中，只有 4%-16%的疣存在多重 HPV 感染相矛盾。该检测方法有资格用于制定未来的基因型特异性疣治疗策略。
