Surentheran T, Harwood C A, Spink P J, Sinclair A L, Leigh I M, Proby C M, McGregor J M, Breuer J
Department of Academic Dermatology, Royal Hospitals NHS Trust, London, UK.
J Clin Pathol. 1998 Aug;51(8):606-10. doi: 10.1136/jcp.51.8.606.
To develop a unified diagnostic approach for the detection of human papillomavirus (HPV) DNA in skin and mucosal biopsies from both immunocompetent and immunosuppressed individuals using a degenerate polymerase chain reaction (PCR) method.
The sensitivity and specificity of three published degenerate primer sets (HVP2/B5 and F14/B15; MY09/MY11; CP62/69 outer and CP65/68 nested primer pairs) were evaluated in PCR reactions with serial dilutions of 12 representative cloned HPV types. This combination of primers was then used to detect HPV DNA in 49 benign and malignant lesions of cutaneous and mucosal origin from immunosuppressed, immunocompetent, and epidermodysplasia verruciformis (EV) patients, and compared with detection rates using single primer sets alone.
The observed sensitivity of MY09/MY11 and CP62/69 + CP65/68 was high for mucosal and EV HPV types, respectively. The sensitivity of all primer sets for cutaneous types was low, but nonetheless the use of this combination of primers allowed HPV DNA detection in all of the benign warts analysed. Several mixed infections were also identified. A high prevalence of HPV DNA was similarly detected in squamous cell carcinomas from immunocompromised patients; the HPV types found were exclusively EV related.
The use of a combined degenerate primer PCR approach considerably improves HPV DNA detection over individual primer sets and allows detection of mixed infections. The findings may help explain the discrepancies in published reports relating to HPV DNA detection in benign and malignant skin lesions. Further modifications to this method are in progress which should significantly improve comprehensive HPV detection and typing for diagnostic purposes.
运用简并聚合酶链反应（PCR）方法，开发一种统一的诊断方法，用于检测免疫功能正常和免疫抑制个体的皮肤及黏膜活检组织中的人乳头瘤病毒（HPV）DNA。
在PCR反应中，使用12种具有代表性的克隆HPV类型的系列稀释液，评估三种已发表的简并引物组（HVP2/B5和F14/B15；MY09/MY11；CP62/69外部引物对和CP65/68巢式引物对）的敏感性和特异性。然后，使用该引物组合检测免疫抑制、免疫功能正常和疣状表皮发育不良（EV）患者的49例皮肤和黏膜来源的良性和恶性病变中的HPV DNA，并与单独使用单个引物组的检测率进行比较。
观察到MY09/MY11和CP62/69 + CP65/68分别对黏膜和EV型HPV具有较高的敏感性。所有引物组对皮肤型HPV的敏感性较低，但尽管如此，使用该引物组合仍能在所有分析的良性疣中检测到HPV DNA。还鉴定出了几种混合感染。在免疫功能低下患者的鳞状细胞癌中同样检测到HPV DNA的高患病率；所发现的HPV类型均与EV相关。
与单个引物组相比，使用简并引物PCR联合方法可显著提高HPV DNA的检测率，并能检测混合感染。这些发现可能有助于解释关于良性和恶性皮肤病变中HPV DNA检测的已发表报告中的差异。目前正在对该方法进行进一步改进，这应能显著提高用于诊断目的的HPV综合检测和分型。
