Forslund Ola, Antonsson Annika, Nordin Peter, Stenquist Bo, Göran Hansson Bengt
Department of Medical Microbiology, Clinical Virology Section, University of Lund, University Hospital, S-205 02 Malmö, Sweden1.
Departments of Dermatology2 and Surgery3, Lundby Hospital, Gothenburg, Sweden.
J Gen Virol. 1999 Sep;80 ( Pt 9):2437-2443. doi: 10.1099/0022-1317-80-9-2437.
A pair of degenerate PCR primers (FAP59/64) was designed from two relatively conserved regions of the L1 open reading frame of most human papillomaviruses (HPV). The size of the generated amplicon was about 480 bp. PCR using these primers was found capable of amplifying DNA from 87% (65/75) of the HPV types tested, its sensitivity being 1-10 copies for HPV-5, -20 and -30 clones. HPV was found in 63% (5/8) of tumour samples and in 63% (5/8) of normal skin biopsies from patients with various cutaneous tumours. HPV-5, HPV-8, HPV-12, HPVvs20-4 and six putatively novel HPV types were identified. No correlation was found to exist between specific HPV and tumour types. Skin surface swab samples from one or more sites on three of four healthy volunteers were found to contain HPV, types 12 and 49 being identified, as well as eight novel HPV types, two of which were also found among the patients. In all, HPV was detected in 75% (9/12) of those tested, five HPV types and 12 novel candidate types being identified, and 37% (7/19) of HPV-positive samples were found to manifest more than one HPV type. All the HPV detected manifested high degrees of nucleotide sequence similarity with HPV types associated with skin lesions and epidermodysplasia verruciformis. The overall HPV finding in the skin samples was 50% (20/40) using the FAP primers as compared to 18% (7/40) using another PCR test designed for skin types. The results thus suggest the new method to be sensitive and generally applicable for detecting cutaneous HPV.
从大多数人乳头瘤病毒(HPV)的L1开放阅读框的两个相对保守区域设计了一对简并PCR引物(FAP59/64)。产生的扩增子大小约为480 bp。发现使用这些引物进行的PCR能够扩增所测试的87%(65/75)的HPV类型的DNA,其对HPV-5、-20和-30克隆的灵敏度为1-10个拷贝。在63%(5/8)的肿瘤样本和63%(5/8)的来自各种皮肤肿瘤患者的正常皮肤活检样本中发现了HPV。鉴定出了HPV-5、HPV-8、HPV-12、HPVvs20-4以及六种可能的新型HPV类型。未发现特定HPV与肿瘤类型之间存在相关性。在四名健康志愿者中的三名志愿者的一个或多个部位的皮肤表面拭子样本中发现含有HPV,鉴定出了12型和49型,以及八种新型HPV类型,其中两种也在患者中发现。总体而言,在75%(9/12)的受测者中检测到了HPV,鉴定出了五种HPV类型和12种新型候选类型,并且发现37%(7/19)的HPV阳性样本表现出不止一种HPV类型。所有检测到的HPV与与皮肤病变和疣状表皮发育不良相关的HPV类型表现出高度的核苷酸序列相似性。使用FAP引物在皮肤样本中的总体HPV检出率为50%(20/40),而使用另一种针对皮肤类型设计的PCR检测方法的检出率为18%(7/40)。因此,结果表明该新方法灵敏且普遍适用于检测皮肤HPV。