Seven biopsies from different carcinomas of the tongue were analyzed for human papilloma virus (HPV) sequences by Southern blot analysis. After hybridization with various types of human papilloma viruses, 3 tumors were found to be positive. Whereas DNA from one tumor hybridized with HPV 2 DNA under conditions of high stringency, the 2 other positive biopsies contained HPV 16 DNA. All positive tumors revealed high copy numbers of the respective DNA. The cleavage pattern of the HPV-2-positive tumor differed from the established HPV 2 prototypes. Minor differences from the HPV 16 prototype were also noted in one of the HPV-16-positive tongue carcinomas.

Seven novel human papillomavirus (HPV) types were isolated and characterized. HPV 94 is related most closely to HPV 10 and belongs to the genus Alphapapillomavirus, whereas HPV 98, HPV 99, HPV 100, HPV 104, HPV 105 and HPV 113 all belong to the genus Betapapillomavirus. These HPV types were isolated from and demonstrated in cutaneous tissue, but HPV 98, HPV 100, HPV 104 and HPV 113 were also detected in malignant oesophageal and oral lesions. The general prevalence of these HPV types in lesions is infrequent.The isolation and characterization of papillomavirus DNA from skin and genital warts revealed the existence of distinct human papillomavirus (HPV) types (zur Hausen et al., 1974;Orth et al., 1977). The large number of HPV types associated with mucosal lesions (reviewed by zur Hausen, 2006) and with the hereditary disease epidermodysplasia verruciformis (Orth, 1986) subsequently confirmed the diversity of the virus family Papillomaviridae. The initial rapid identification and characterization of HPV types associated with genital/mucosal lesions focused attention mainly on the large number of HPV types in the genus Alphapapillomavirus (de Villiers et al., 2004a). Availability of DNA-amplification methods facilitated the demonstration of HPV DNA in cutaneous lesions. Several subsequent studies attempted to associate specific HPV types with the aetiology of non-melanoma skin cancer in immunosuppressed and immunocompetent patients (Shamanin et al., 1994(Shamanin et al., , 1996 Berkhout et al., 1995Berkhout et al., , 2000de Villiers et al., 1997;Forslund et al., 1999Forslund et al., , 2003bForslund et al., , 2007Asgari et al., 2008;Harwood et al., 2000Harwood et al., , 2004, whereas others demonstrated the presence of HPV in and on normal skin (Astori et al., 1998;Antonsson et al., 2000;Hazard et al., 2007;Chen et al., 2008). These studies all resulted in the identification of a vast spectrum of additional putative novel HPV types in the form of PCRamplified partial open reading frame (ORF) L1 sequences (Shamanin et al., 1996;de Villiers et al., 1997; Astori et al., 1998;Antonsson et al., 2000;Forslund et al., 2003a;.Full-length genomes of a relatively small number of these HPV isolates have been cloned and characterized (Delius et al., 1998;Forslund et al., 2003a;Vasiljevic et al., 2007Vasiljevic et al., , 2008. Definition of a papillomavirus type requires the isolation and characterization of a full-length genome in order to verify the genome organization characteristic of papillomaviruses (de Villiers et al., 2004a). We report here the isolation and characterization of seven novel HPV types, all of which were initially identified as partial ORF L1 sequences.The complete genomes of seven novel HPV types were amplified through rolling-circle amplification of cellular DNA and subsequent long PCR amplification (Leppik et al., 2007), using primers that had been designed on the respective partial L1 sequence on which the respective putative HPV types had initially been identified (de Villiers et al., 1997; Astori et...

32P-labelled cloned HPV 6 DNA was used as probe to analyze human genital tumors for DNA sequences homologous to HPV 6 DNA. Ninety three percent of all condylomata acuminata (41 out of 44) were found to harbor HPV 6 DNA. Of the remaining three, one contained HPV 1 DNA. No papillomavirus DNA was identified in the two other tumors. All three invasively growing giant condylomata acuminata (Buschke-Löwenstein tumors) investigated also contained HPV 6 DNA. Two out of six atypical condylomata of the cervix hybridized with HPV 6 DNA under stringent conditions, one only under conditions of low stringency. All DNA preparations from malignant tumors studies (54 cervical carcinomas, 10 penile carcinomas, two vulvar carcinomas) failed to anneal with HPV 6 DNA, even under conditions of low stringency. Although all HPV 6-positive condylomata acuminata analyzed in this study revealed HPV 6 DNA of regular molecular weight (5.1 x 10(6)), two of the Buschke-Löwenstein tumors, as well as one of the two positive atypical condylomata of the cervix, contained HPV 6 DNA with a remarkable size classes occurred in a supercoiled form without evidence for integration into host cell DNA.