Human papillomavirus genotyping using next generation sequencing (NGS) in cervical lesions: Genotypes by histologic grade and their relative proportion in multiple infections.
Human papillomavirus genotyping using next generation sequencing (NGS) in cervical lesions: Genotypes by histologic grade and their relative proportion in multiple infections.
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DOI：
10.1371/journal.pone.0278117
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发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
人乳头瘤病毒 (HPV) 的敏感和特异性基因分型对于疫苗有效性的监测和监测至关重要。在这里，使用巢式 PCR 和下一代测序 (NGS) 技术在 137 个具有不同组织学的宫颈样本（79 个≤CIN1 和 58 个 CIN3+）中鉴定了 HPV 基因型，并计算了多种感染中每种基因型的相对比例。所有样品均已预先通过 PCR 反向印迹杂交 (PCR-RBH) 进行基因分型，从而可以对两种技术之间进行一致性分析。 85% 的 ≤CIN1 病例存在多重感染，而 CIN3+ 病例中只有 41% 存在多重感染 (p<0.001)。考虑到低风险 (LR-) 和高风险 (HR-) HPV 基因型，在 ≤CIN1 病例中观察到显着的基因型多样性；而在 CIN3+ 中，多样性较低，大多数情况下 HR-HPV 占主导地位，尤其是 HPV16。此外，CIN3+病例中HR-HPV基因型在每个样本中所占的比例中占主导地位[(HPV16 (62.5%)，其次是HPV31和HPV58(各8.3%)]，高于≤CIN1病例[(HPV16 (17.7%)，其次是HPV52 (14.7%)和HPV31 (10.3%)]。所有基因型的 PCR-RBH 和 NGS 均高于 90%（总体 Kappa 为 0.7），尽管 NGS 鉴定出 PCR-RBH 未检测到的 89 个 HPV 基因型阳性结果，证明其具有更高的敏感性，这些结果表明基因型多样性的减少和/或多重感染中 HR-HPV 相对比例的增加可被视为恶性进展潜在风险的生物标志物。
Sensitive and specific genotyping of human papillomaviruses (HPVs) is critical for the surveillance and monitoring of the vaccine effectiveness. Here, HPV genotypes were identified in 137 cervical samples with different histology (79 ≤CIN1 and 58 CIN3+) using Nested-PCR followed by Next-Generation sequencing (NGS) and relative proportions for each genotype in multiple infections were computed. All samples had been previously genotyped by PCR-Reverse Blotting Hybridization (PCR-RBH) thus allowing for a concordance analysis between both techniques. Multiple infections were present in 85% of ≤CIN1 cases compared to only 41% in CIN3+ cases (p<0.001). Among ≤CIN1 cases a towering genotypic diversity was observed, considering both low (LR-) and high risk (HR-) HPV genotypes; while among CIN3+, diversity was lower, HR-HPVs prevailing in most cases, especially HPV16. Furthermore, the predominance of HR-HPV genotypes in the proportions identified in each sample was higher in CIN3+ cases [(HPV16 (62.5%), followed by HPV31 and HPV58 (8.3% each)], than in ≤CIN1 cases [(HPV16 (17.7%), followed by HPV52 (14.7%) and HPV31 (10.3%)]. Agreement between PCR-RBH and NGS was higher than 90% for all genotypes (with an overall Kappa of 0.7), even though NGS identified eighty-nine positive results for HPV genotypes that had not been detected by PCR-RBH, evidencing its greater sensitivity. These results suggest that a reduction in genotypic diversity and/or an increase in the relative proportion of HR-HPVs in multiple infections can be considered as a biomarker for the potential risk of malignant progression.
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影响因子：
51.1
作者：
de Sanjose, Silvia;Quint, Wim G. V.;Xavier Bosch, F.
通讯作者：
Xavier Bosch, F.
影响因子：
9.4
作者：
Gravitt, PE;Peyton, CL;Wheeler, CM
通讯作者：
Wheeler, CM
影响因子：
6.4
作者：
De Vuyst, Hugo;Clifford, Gary M.;Franceschi, Silvia
通讯作者：
Franceschi, Silvia
影响因子：
8.8
作者：
Bosch, F. X.;Castellsague, X.;de Sanjose, S.
通讯作者：
de Sanjose, S.
影响因子：
3.4
作者：
Cuschieri, KS;Cubie, HA;McGoogan, E
通讯作者：
McGoogan, E