Detection of human papillomavirus DNA in extragenital Bowen's disease by using a nested PCR / 中华皮肤科杂志

ObjectiveTo develop a new nested PCR to detect human papillomavirus (HPV) DNA in lesions of extragenital Bowen's disease.Methods DNA was extracted from the lesions of 41 patients with extragenital Bowen's disease and suhjected to the amplification of HPV by a nested PCR.Five primers,including CN1FR,CN2FR,CN3FR,CN4FR and CN5FR,were designed and used in the second round PCR.ResultsBased on the ClustalX analysis,69 HPV subtypes,including mucosal types,cutaneous types and epidermodysplasia verruciformis-associated types,could be amplified by using the 5 designed primers.The detection limit varied from 10-3 to 10-2 copies of HPV DNA for this PCR.Of the 41 lesional specimens,5 were positive for HPV DNA,including 3 cases of high-risk HPV types(2 cases of HPV 16,1 case of HPV 33) and 2 cases of cutaneous HPV types(1 case of HPV 27 and 1 case of HPV 76).ConclusionsThe improved nested PCR is highly sensitive and specific for the detection of HPV DNA in lesions of extragenital Bowen's disease.The development of extragenital Bowen's disease may be associated with the infection with high-risk mucosal HPV types in some patients.

Relationship between human papillomavirus and extragenital Bowen's disease / 中华皮肤科杂志

Objective To assess the relationship between human papillomavirus (HPV) and extrageni-tal Bowen's disease. Methods Regular PCR with consensus primers for LI region as well as mix primers and nested PCR were performed to detect the DNA of a broad range of cutaneous and mucosal HPV types in tissue samples from lesions of 41 patients with extragenital Bowen's disease and from normal skin of 48 human controls. Semiquantitative PCR and tyramide-based in situ hybridization (ISH) were also conducted to determine the load and localization of HPV DNA in HPV-positive samples. Results HPV DNA was detected in lesions from 5 (12%) of the 41 patients with extragenital Bowen's disease. Of the 5 HPV-positive patients, 3 carried mucosal HPV types (HPV16 in 2 cases, HPV 33 in 1 case) with a viral load of 101 to 103 copies, 2 cutaneous HPV types (HPV27 in 1 case and HPV76 in 1 case). As ISH showed, there was a generalized expression of mucosal HPV DNA in most tumor cell nuclei but not in peritumoral normal tissue, and no expression of cutaneous HPV DNA was observed in lesions. HPV DNA was detected in 1 (2.1%) control tissue sample, which proved to be epidermodysplasia verruciformis-associated HPV23. There was no significant difference in the detection rate of HPV DNA between the patients and controls. The viral load of cutaneous HPV types amounted to 10-2 to 10-3 copies in the 2 patients, which was similar to that of HPV 23 in the normal control. Conclusion Mucosal HPV types may be closely associated with the development of extragenital Bowen's disease.