Development of a multiplex PCR method for detecting and typing human papillomaviruses in verrucae vulgaris.

ABSTRACT

The methods for detecting and typing human papillomavirus (HPV) in most molecular epidemiological surveys of verrucae vulgaris were based on PCR followed by sequencing or hybridization. However, the amplification efficacies of different assays for the detection of HPV DNAs varied largely. In this study, a novel multiplex PCR method to detect and type the HPVs (HPV-1, -2, -27 and -57) related to verrucae vulgaris was described. This method allows detecting and typing HPV DNA simultaneously in one reaction based on the length of the PCR products after electrophoresis. The sensitivity and specificity of this multiplex PCR method was assessed with the standard template panels and the spiking sample panels, and evaluated with the clinical samples, compared with PCR assay with primer MY09/11. The results showed the novel method had reliable clinical sensitivity (97.6%) and specificity (100%), significantly higher than that of the PCR using consensus primer, MY09/11. In addition, this method can effectively detect multiple HPV infection within the lesions. This simplified, economic and time-saving multiplex PCR method provides a useful additional tool for the clinical epidemiological study of verrucae vulgaris.