PCR detection of human papillomavirus: comparison between MY09/MY11 and GP5+/GP6+ primer systems

TLDR

Human papillomavirus is the leading cause of cervical cancer and the most common sexually transmitted infection in women, and PCR amplification is the most sensitive method for detecting cervicovaginal HPV, though primer system sensitivities vary and must be considered when comparing studies. The study compared MY09/MY11 and GP5+/GP6+ primer sets by testing 208 cervicovaginal lavage samples with probes for 39 HPV types. Both primer sets detected a broad range of HPV genotypes with comparable overall prevalences (45% vs 43%), yet MY-PCR identified more multiple infections and differed in detecting types 35, 53, and 61, as confirmed by serial dilution showing a 3‑log reduction for type 35 by MY-PCR and for types 53 and 61 by GP+-PCR.

Abstract

Human papillomavirus (HPV) is an etiologic agent of cervical cancer and is the most common sexually transmitted disease in women. PCR amplification of HPV genomes is the most sensitive method for the detection of cervicovaginal HPV. We have compared the two most commonly used PCR primer sets, MY09/MY11 (MY-PCR) and GP5+/GP6+ (GP+-PCR), for the detection of HPV DNA in cervicovaginal lavage samples from 208 women. Oligonucleotide probes for 39 different HPV types were used. Both primer sets amplified a wide spectrum of HPV genotypes and detected similar overall prevalences of 45% (94 of 208) and 43% (89 of 208), respectively. The MY-PCR system detected 27 of 30 (90%) samples with multiple HPV types, whereas the GP+-PCR system detected 14 of 30 (47%) samples with multiple HPV types. Differences in the detection of HPV types 35, 53, and 61 were noted between the two primer systems. Serial dilution of plasmid templates indicated a 3-log decrease in the amplification of HPV type 35 by MY-PCR and HPV types 53 and 61 by GP+-PCR. These results indicate that although the MY-PCR and GP+-PCR identified nearly equivalent prevalences of HPV in a set of clinical samples, differences in the detection of specific types and infections with multiple types were found. Differences in the sensitivities and characteristics of the PCR systems for the detection of HPV within clinical samples should be considered when comparing data between studies and/or in designing new studies or clinical trials.