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Evidence for missing HPV-45 and -59 positives with the SPF10-DEIA-LiPA25 (version 1) platform compared to the type-specific qPCR assays and the impact on vaccine effectiveness estimates.
Journal of Clinical Microbiology ( IF 6.1 ) Pub Date : 2020-10-21 , DOI: 10.1128/jcm.01626-20
Kahren van Eer 1 , Suzan Leussink 1 , Tim T Severs 1 , Naomi van Marm-Wattimena 1 , Petra J Woestenberg 1 , Johannes A Bogaards 1, 2 , Audrey J King 3
Affiliation  

Human papillomavirus (HPV) epidemiological and vaccine studies require highly sensitive HPV detection systems. The widely used broad-spectrum SPF10-DEIA-LiPA25 (SPF10 method) has reduced sensitivity toward HPV-45 and -59. Therefore, anogenital samples from the PASSYON study were retrospectively analyzed with type-specific (TS) HPV-45 and -59 real-time quantitative PCR (qPCR) assays. The SPF10 method missed 51.1% of HPV-45 and 76.1% of HPV-59 infections that were detected by the TS qPCR assays. The viral copy number (VCn) of SPF10-missed HPV-45 and -59 was significantly lower than SPF10-detected HPV-45 and -59 (P < 0.0001 for both HPV types). Sanger sequencing showed no phylogenetic distinction between SPF10-missed and SPF10-detected HPV-59 variants, but variants bearing the A6562G single-nucleotide polymorphism (SNP) in the SPF10 target region were more likely to be missed (P = 0.0392). HPV cooccurrence slightly influenced the detection probability of HPV-45 and -59 with the SPF10 method. Moreover, HPV-59 detection with the SPF10 method was hampered more in nonvaccinated women than vaccinated women, likely due to a stronger masking effect by increased HPV cooccurrence in the former group. Consequently, the SPF10 method led to a strong negative vaccine effectiveness (VE) of –84.6% against HPV-59, while the VE based on TS qPCR was 3.1%. For HPV-45, the relative increase in detection in nonvaccinated women compared vaccinated women was more similar, resulting in comparable VE estimates. In conclusion, this study shows that HPV-45 and -59 detection with the SPF10 method is dependent on factors including VCn, HPV cooccurrence, and vaccination, thereby showing that knowledge of the limitations of the HPV detection method used is of great importance.

中文翻译:

与特定类型的qPCR分析相比,使用SPF10-DEIA-LiPA25(版本1)平台缺失HPV-45和-59阳性的证据,以及对疫苗有效性估计的影响。

人乳头瘤病毒(HPV)流行病学和疫苗研究需要高度敏感的HPV检测系统。广泛使用的广谱SPF 10 -DEIA-LiPA 25(SPF 10方法)对HPV-45和-59的敏感性降低。因此,使用类型特异性(TS)HPV-45和-59实时定量PCR(qPCR)分析对PASSYON研究中的肛门生殖器样本进行了回顾性分析。通过TS qPCR分析检测到的SPF 10方法漏掉了51.1%的HPV-45和76.1%的HPV-59感染。SPF 10缺失的HPV-45和-59的病毒拷贝数(VCn)明显低于SPF 10缺失的HPV-45和-59(P对于两种HPV类型,均<0.0001)。Sanger测序显示在SPF 10缺失和SPF 10检测到的HPV-59变体之间没有系统发育差异,但是在SPF 10目标区域中带有A6562G单核苷酸多态性(SNP)的变体更容易被遗漏(P = 0.0392) 。HPV的同时出现对使用SPF 10方法检测HPV-45和-59的检测概率有轻微影响。此外,用SPF 10方法检测HPV-59在未接种疫苗的女性中比在接种疫苗的女性中受阻更大,这可能是由于前一组中增加的HPV并发掩盖作用更强。因此,SPF 10该方法导致针对HPV-59的强阴性疫苗效力(VE)为–84.6%,而基于TS qPCR的VE为3.1%。对于HPV-45,与未接种疫苗的妇女相比,未接种疫苗的妇女的检出率相对增加更为相似,因此VE估算值可比。总之,这项研究表明,使用SPF 10方法检测HPV-45和-59取决于VCn,HPV共存和接种疫苗等因素,因此表明了解所用HPV检测方法的局限性非常重要。
更新日期:2020-10-27
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