PCR detection of human papillomavirus of the mucosa: comparison between MY09/11 and GP5+/6+ primer sets.
OBJECTIVE
Human papillomaviruses (HPV) are the causal agent for the development of carcinomas in the cervix uteri and further pathological changes of the skin including mucosa, particularly warts, condylomas and dysplasias. Therefore, we investigated the efficacy of different consensus primers pairs for HPV detection by PCR using brushed samples from the oral cavity in comparison with samples from the cervix uteri.


METHODS
In the present study, we used two well-established sets of PCR primers in different combinations for the detection of HPV DNA in 106 non-invasive brush biopsy samples of the oral mucosa and 56 samples from the cervix uteri. Direct sequencing of PCR products in all cases determined HPV genotype and specificity.


RESULTS
Overall, HPV was detected in 69 of 106 oral mucosa samples. HPV specific amplicons were obtained in 35.8% (N = 38) when using GP5+/6+ primers. The positivity rate was increased to 65.1% in a GP5+/6+ auto-nested PCR approach. In contrast, MY9/11 PCR and nested PCR with MY9/11 outer followed by GP5+/6+ inner primers yielded 2.2% and 16.1%, respectively. In gynaecological samples, PCR results were similar independent of the primer combination used. Thus, DNA quality and DNA content could be additional factors influencing the rate of positivity.


CONCLUSIONS
For oral mucosa samples, auto-nested GP5+/6+ PCR is in our hands the most suitable approach for epidemiological studies because of its high sensitivity, high reliability and reproducibility as well as its relatively simple laboratory procedure.
Remmerbach
Torsten W
TW
Department of Oral, Maxillofacial and Facial Plastic Surgery, University of Leipzig, Leipzig, Germany.
Brinckmann
Ute G
UG
Hemprich
Alexander
A
Chekol
M
M
Kühndel
K
K
Liebert
Uwe Gerd
UG
D001706:Biopsy / Q000379:methods; D002294:Carcinoma, Squamous Cell / Q000821:virology*; D002584:Cervix Uteri / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis / Q000302:isolation & purification; D005260:Female; D006801:Humans; D009061:Mouth Mucosa / Q000821:virology*; D009062:Mouth Neoplasms / Q000821:virology*; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000821:virology
D017931:DNA Primers; D004279:DNA, Viral