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Use of Multiplex Polymerase Chain Reaction for Detection of High-Risk Human Papillomavirus Genotypes in Women Attending Routine Cervical Cancer Screening in Harare.
Intervirology ( IF 4.6 ) Pub Date : 2019-08-14 , DOI: 10.1159/000502206
Takudzwa Marembo 1 , Racheal Dube Mandishora 2 , Margaret Borok 3
Affiliation  

BACKGROUND/AIMS In Zimbabwe, cervical cancer is screened through cytology and visual inspection with acetic acid and cervicography (VIAC). The effectiveness of these methods can be increased if complemented by a human papillomavirus DNA detection tool since most cervical cancer cases are caused by persistent infection with high-risk human papillomavirus (HR-HPV) genotypes. Moreover, the possibility of multiple-genotype HR-HPV infections warrants the need for HPV detection tools with the capacity to detect both single and multiple infections. The aim of this study was to detect HR-HPV genotypes (HPV 16, 18, 31, 33, 35, 45, 51, 52, 56, and 58), using multiplex polymerase chain reaction (PCR), in stored cervicovaginal swabs from both HIV-positive and HIV-negative women reporting for routine cervical cancer screening. METHODOLOGY Stored cervicovaginal swabs from sexually active women who underwent VIAC at the Parirenyatwa Referral Hospital in Harare, Zimbabwe, between February and April 2015 and had received HIV counselling and testing were genotyped for the selected 10 HR-HPV genotypes using in-house multiplex PCR. The results from the multiplex PCR were compared to those previously obtained when the same samples were HPV genotyped with next-generation sequencing (NGS) on an MiSeq platform (Illumina; USA). RESULTS A total of 136 women were recruited and all 10 HR-HPV genotypes were detected. Quality control failed in 3 of the 136 swabs during the multiplex PCR reactions. The prevalence of HR-HPV genotypes in the study subjects was 53% (70/133). HIV-infected women were 1.67 times more likely to be infected with HR-HPV than were HIV-negative women (OR 1.67; p = 0.17). Of the 70 HR-HPV-positive cases, 37% (26/70) had multiple HR-HPV infections, and the majority of them were HIV infected. HIV-infected women were 1.86 times more likely to have multiple HR-HPV infections than HIV-negative women (OR 1.86; p = 0.20). Multiplex PCR and NGS had an almost perfect concordance rate in -HR-HPV detection (κ = 0.960), with only 3 discordant cases (negative with NGS and positive for HPV16 with multiplex PCR). CONCLUSION Multiplex PCR can detect HR-HPV genotypes that are common in Zimbabwe and could be used to detect HR-HPV genotypes from women attending cervical cancer screening programs at the Parirenyatwa VIAC clinic in Harare.

中文翻译:

使用多重聚合酶链反应检测在哈拉雷参加常规宫颈癌筛查的女性的高危人乳头瘤病毒基因型。

背景/目的 在津巴布韦,宫颈癌是通过细胞学和醋酸肉眼检查和宫颈造影 (VIAC) 来筛查的。如果辅以人乳头瘤病毒 DNA 检测工具,可以提高这些方法的有效性,因为大多数宫颈癌病例是由高危人乳头瘤病毒 (HR-HPV) 基因型的持续感染引起的。此外,多基因型 HR-HPV 感染的可能性需要具有检测单一和多重感染能力的 HPV 检测工具。本研究的目的是使用多重聚合酶链反应 (PCR) 检测储存的宫颈阴道拭子中的 HR-HPV 基因型(HPV 16、18、31、33、35、45、51、52、56 和 58) HIV 阳性和 HIV 阴性的妇女都报告进行常规宫颈癌筛查。方法学 对 2015 年 2 月至 4 月在津巴布韦哈拉雷的 Parirenyatwa 转诊医院接受 VIAC 并接受 HIV 咨询和检测的性活跃女性的宫颈阴道拭子进行基因分型,使用内部多重 PCR 对选定的 10 种 HR-HPV 基因型进行基因分型。将多重 PCR 的结果与先前在 MiSeq 平台(Illumina;美国)上使用下一代测序 (NGS) 对相同样本进行 HPV 基因分型时获得的结果进行比较。结果 共招募了 136 名女性,检测了所有 10 种 HR-HPV 基因型。在多重 PCR 反应期间,136 个拭子中有 3 个质量控制失败。研究对象中 HR-HPV 基因型的流行率为 53% (70/133)。感染 HIV 的女性感染 HR-HPV 的可能性是 HIV 阴性女性的 1.67 倍(OR 1.67;p = 0.17)。70例HR-HPV阳性病例中,37%(26/70)为HR-HPV多重感染者,其中以HIV感染者居多。感染 HIV 的女性感染 HR-HPV 的可能性是 HIV 阴性女性的 1.86 倍(OR 1.86;p = 0.20)。多重 PCR 和 NGS 在 -HR-HPV 检测中具有几乎完美的一致性率(κ = 0.960),只有 3 个不一致的病例(NGS 阴性,多重 PCR HPV16 阳性)。结论 多重 PCR 可以检测津巴布韦常见的 HR-HPV 基因型,并可用于检测在哈拉雷的 Parirenyatwa VIAC 诊所参加宫颈癌筛查计划的女性的 HR-HPV 基因型。感染 HR-HPV 的可能性是 HIV 阴性女性的 86 倍 (OR 1.86; p = 0.20)。多重 PCR 和 NGS 在 -HR-HPV 检测中具有几乎完美的一致性率(κ = 0.960),只有 3 个不一致的病例(NGS 阴性,多重 PCR HPV16 阳性)。结论 多重 PCR 可以检测津巴布韦常见的 HR-HPV 基因型,并可用于检测在哈拉雷的 Parirenyatwa VIAC 诊所参加宫颈癌筛查计划的女性的 HR-HPV 基因型。感染 HR-HPV 的可能性是 HIV 阴性女性的 86 倍 (OR 1.86; p = 0.20)。多重 PCR 和 NGS 在 -HR-HPV 检测中具有几乎完美的一致性率(κ = 0.960),只有 3 个不一致的病例(NGS 阴性,多重 PCR HPV16 阳性)。结论 多重 PCR 可以检测津巴布韦常见的 HR-HPV 基因型,并可用于检测在哈拉雷的 Parirenyatwa VIAC 诊所参加宫颈癌筛查计划的女性的 HR-HPV 基因型。
更新日期:2019-11-01
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