Evaluation of a Novel Broad-Spectrum PCR-Multiplex Genotyping Assay for Identification of Cutaneous Wart-Associated Human Papillomavirus Types
Evaluation of a Novel Broad-Spectrum PCR-Multiplex Genotyping Assay for Identification of Cutaneous Wart-Associated Human Papillomavirus Types
复制标题
DOI：
10.1128/jcm.02122-09
复制
发表时间：
2010-05-01
影响因子：
9.4
通讯作者：
Quint, Wim G. V.
中科院分区：
文献类型：
--
作者：
de Koning, Maurits N. C.;ter Schegget, Jan;Quint, Wim G. V.
大量的人乳头瘤病毒（HPV）类型，分布在五个乳头瘤病毒属中，可在皮肤中检测到。在皮肤疣中检测到属于α、γ和μ属的HPV类型。虽然疣是一种非常普遍的皮肤疾病，特别是在儿童（33%）和器官移植受者（45%）中，但目前还不存在针对这些皮肤疣相关HPV类型的最先进HPV基因分型检测。皮肤疣再次成为人们关注的焦点，因为随着慢性免疫抑制患者数量的增加，其临床相关性也在增加。本研究的目的是使用PCR和Luminex xMAP技术开发和评估所有已知皮肤疣相关HPV类型的基于DNA的基因分型系统。广谱PCR扩增了所有已知疣相关HPV类型的DNA，包括α（HPV 2、3、7、10、27、28、29、40、43、57、77、91和94）、γ（HPV 4、65、95、48、50、60和88）、μ（HPV 1和63）和nu（HPV 41）。使用质粒HPV DNA和一组45个先前表征的皮肤疣活检标本显示出高特异性的探针进行了评估。在100份非生殖器皮肤疣拭子中，96%也发现了HPV。HPV 1、2、27和57型是89%拭子中检出的最常见HPV类型。总之，这种基于Luminex的基因分型系统可识别所有已知的皮肤疣HPV类型，包括遗传学相关类型，具有高度HPV类型特异性，适用于大规模流行病学研究。
A large number of human papillomavirus (HPV) types, distributed over five papillomavirus genera, are detectable in the skin. HPV types belonging to the alpha, gamma, and mu genera have been detected in cutaneous warts. A state-of-the-art HPV genotyping assay for these cutaneous wart-associated HPV types does not exist although warts constitute a highly prevalent skin condition, especially in children (33%) and organ transplant recipients (45%). Cutaneous warts are again the focus of attention as their clinical relevance rises with the increasing number of chronically immunosuppressed patients. The objective of this study was to develop and evaluate a DNA-based genotyping system for all known cutaneous wart-related HPV types using PCR and Luminex xMAP technology. The broad-spectrum PCR amplified DNA of all known wart-associated HPV types from the genera alpha (HPVs 2, 3, 7, 10, 27, 28, 29, 40, 43, 57, 77, 91, and 94), gamma (HPVs 4, 65, 95, 48, 50, 60, and 88), mu (HPVs 1 and 63), and nu (HPV41). The probes were evaluated using plasmid HPV DNA and a panel of 45 previously characterized cutaneous wart biopsy specimens showing high specificity. HPV was also identified in 96% of 100 swabs from nongenital cutaneous warts. HPV types 1, 2, 27, and 57 were the most prevalent HPV types detected in 89% of the swabs. In conclusion, this Luminex-based genotyping system identifies all known cutaneous wart HPV types including phylogenetically related types, is highly HPV type specific, and is suitable for large-scale epidemiological studies.