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Journal of Clinical Microbiology, March 2009, p. 541-546, Vol. 47, No. 3
0095-1137/09/$08.00+0     doi:10.1128/JCM.02007-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Modified General Primer PCR System for Sensitive Detection of Multiple Types of Oncogenic Human Papillomavirus{triangledown}

Anna Söderlund-Strand,1,2 Joyce Carlson,3 and Joakim Dillner1,2*

WHO HPV LabNet Global Reference Laboratory, Department of Clinical Microbiology,1 Department of Laboratory Medicine, Lund University, Malmö University Hospital, SE-20502 Malmö, Sweden,2 Department of Clinical Chemistry and Pharmacology, Lund University Hospital, SE-22185 Lund, Sweden3

Received 17 October 2008/ Returned for modification 12 December 2008/ Accepted 6 January 2009

Human papillomavirus (HPV) infection is a necessary cause of cervical cancer and cervical dysplasia. Accurate and sensitive genotyping of multiple oncogenic HPVs is essential for a multitude of both clinical and research uses. We developed a modified general primer (MGP) PCR system with five forward and five reverse consensus primers. The MGP system was compared to the classical HPV general primer system GP5+/6+ using a proficiency panel with HPV plasmid dilutions as well as cervical samples from 592 women with low-grade cytological abnormalities. The reference method (GP5+/6+) had the desirable high sensitivity (five copies/PCR) for five oncogenic HPV types (HPV type 16 [HPV-16], HPV-18, HPV-56, HPV-59, and HPV-66). The MGP system was able to detect all 14 oncogenic HPV types at five copies/PCR. In the clinical samples, the MGP system detected a significantly higher proportion of women with more than two concomitant HPV infections than did the GP5+/6+ system (102/592 women compared to 42/592 women). MGP detected a significantly greater number of infections with HPV-16, -18, -31, -33, -35, -39, -42, -43, -45, -51, -52, -56, -58, and -70 than did GP5+/6+. In summary, the MGP system primers allow a more sensitive amplification of most of the HPV types that are established as oncogenic and had an improved ability to detect multiple concomitant HPV infections.

* Corresponding author. Mailing address: WHO HPV LabNet Global Reference Laboratory, Department of Clinical Microbiology, Lund University, Malmö University Hospital, SE-20502 Malmö, Sweden. Phone: 46 40 338126. Fax: 46 40 337312. E-mail: joakim.dillner{at}med.lu.se

{triangledown} Published ahead of print on 14 January 2009.

Journal of Clinical Microbiology, March 2009, p. 541-546, Vol. 47, No. 3
0095-1137/09/$08.00+0     doi:10.1128/JCM.02007-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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