Establishment of a genotyping method for the detection of 32 types of genital tract human papillomaviruses based on a PCR reverse dot blot array

Objective To establish a genotyping method for the detection of 32 types of genital tract human papillomaviruses( HPV)based on a PCR reverse dot blot( PCR-RDB) assay. Methods HPV genotype-specific primers and probes based on MY09 /11 were designed and synthesized,and a genotyping method based on PCR-RDB was established. Then,the established method was evaluated by HPV vector and used to detect the types of 300 clinical cervical cells samples. The obtained results were compared with those from a commercial PCR-RDB method. Results The established HPV PCR-RDB method had good specificity,accuracy and repeatability.The lowest HPV detection limits were 50 copies / reaction and 5 copies / reaction for 39 and 51 types of HPV and the other types,respectively. The total positive rate,single infection rate and multiple infection rate for 300 clinical samples detected by the established PCRRDB method were 18. 67%( 56 /300),12. 33%( 37 /300) and 6. 33%( 19 /300),respectively,and there was no statistical significance when compared with the results from the commercial PCR-RDB method( χ2values were 0. 744,0. 597 and 0. 118,respectively,and all of P 0. 05). The Kappa value for the consistency test of the two methods was 0. 889( P 0. 01). Conclusion A PCR-RDB method for the genotyping of 32 types of genital tract HPVs is successfully established,which has fine sensitivity,specificity,and repeatability,and is suitable for the detection of clinical samples.