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

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.

High frequency of multiple HPV types detection in Fanconi anemia patients oral swabs

“…In fact, other studies showed that when multiple HPV DNAs with different amplification efficiencies are present in the same sample, selective amplification of one HPV DNA over another can occur. 27,28 The fact that some HPV genotypes that were detected by Sanger 22 sequencing with random primers were not detected by reverse hybridization may be explained, at least in part, by variations during PCR due to differences in primers and probes sets between the two methodologies, when working with multiple HPV infected samples. This could also explain the finding of HPV-type 89 using hybridization, a type that was not identified by Sanger 22 sequencing, and could also justify the differences regarding the number of HPV types found when multiple HPV types are present in the same sample.…”

Reliable detection of Human papillomavirus in recurrent laryngeal papillomatosis and associated carcinoma of archival tissue

“…Broad-spectrum PCRs like GP5þ/6þ and MY09/11 amplify the widely conserved viral gene L1 open reading frame. Both methods are considered to be highly sensitive and specific [Qu et al, 1997]. On the other hand, compared to type-specific PCRs the use of broadspectrum test to detect HPV genotypes may underestimate the prevalence of certain genotypes, due to competition between genotypes in mixed infections [Qu et al, 1997;Molijn et al, 2005;Depuydt et al, 2007].…”

Cervical HPV DNA Detection as a Predictor of a Recurrent SIL Diagnosis Among Untreated Women

“…At enrollment, cervical specimens collected by cervicovaginal lavage were tested for a diverse set of HPV types, followed by individual type determinations, using a HPV DNA-specific assay Hybrid Capture Tube Test (HC) (Digene Corp., Gaithersburg, MD) [4]. Consensus primer PCR assays were performed to increase the sensitivity of HPV DNA detection [5]. We combined the results of the assays such that a positive signal in either or both assays was considered HPV DNA-positive.…”