Journal of Virological Methods 122 (2004) 87–93 Nested PCR with the PGMY09/11 and GP5 + /6 + primer sets improves detection of HPV DNA in cervical samples Andrea L. Fuessel Haws a , Qin He a , Peter L. Rady c , Lifang Zhang b , James Grady b , Thomas K. Hughes a , Kendra Stisser a , Rolf Konig a , Stephen K. Tyring a,c,∗ a Departments of Microbiology/Immunology, University of Texas Medical Branch, Galveston, TX 77030, USA b The Office of Biostatistics, University of Texas Medical Branch, Galveston, TX 77030, USA c Department of Dermatology, University of Texas Medical School at Houston, Houston, TX 77030, USA Received 31 March 2004; received in revised form 4 August 2004; accepted 5 August 2004 Abstract Based on epidemiological and research evidence, HPV has a causal role in cervical carcinogenesis. Several HPV detection methods exist to date; the most commonly used method for detection of genital HPVs consists of nested PCR using the MY09/11 and GP5 + /6 + primer sets (MY/GP + ). Recently, the PGMY09/11 primer set, a modified version of the MY09/11 primer set, was introduced for single PCR and was found to detect a wider range of HPV types. The next logical step was taken and the efficacy of nested PCR using the PGMY09/11 and GP5 + /6 + primer sets (PGMY/GP + ) to detect HPV in cervical samples was evaluated. In this comparative study, nested PCR using the novel PGMY/GP + primer set combination was found to be more type sensitive than the nested PCR with the MY/GP + primer sets, detecting a wider range of HPV types, low copy HPVs, and better characterizing samples infected with multiple strains of HPV. Standardization and use of the PGMY/GP + PCR system could aid physicians in providing more efficient HPV screening and better treatment for patients. © 2004 Elsevier B.V. All rights reserved. Keywords: Human papillomavirus; Detection; PCR; PGMY; GP + 1. Introduction Human papillomavirus (HPV) infection remains a promi- nent concern in both the research and medical fields. It is capable of infecting the basal epithelial cells of all men and women worldwide, regardless of age or race, and can cause a wide variety of clinical manifestations, which can be either benign or malignant (Burd, 2003). Although HPV has been associated with many diseases, cervical cancer has particu- lar significance, being the second most common cancer in women worldwide, the third most common cancer in women in the United States, and the principal cancer of women in most developing countries (Burd, 2003; Munoz et al., 2003). ∗ Corresponding author. Tel.: +1 713 528 8818; fax: +1 281 335 4605. E-mail address: stephen.k.tyring@uth.tmc.edu (S.K. Tyring). More than 230 HPV types have been recognized to date on the basis of DNA sequence data showing genomic differences; 85 HPV genotypes are fully characterized (Burd, 2003; de Villiers, 1999). There are approximately 40 known genital HPVs, which are further divided into high-risk (HR-HPV; oncogenic: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, 82; probably carcinogenic: 26, 53, 66) and low risk (LR-HPV; e.g. 6, 11, 40, 42, 43, 44, 54, 61, 70, 72, 81, and candHPV89/CP6108) groups for cervical cancer (Terai and Burk, 2002; Munoz et al., 2003). HPV type 16 is the most common HPV type detected in cervical cancers worldwide, followed by HPV 18 (Munoz, 2000). Several PCR based HPV detection methods exist to date; however the most commonly used PCR methods for the detection of genital HPVs have used the MY09/11 and GP5 + /6 + primer sets (MY/GP + )(Strauss et al., 2000; 0166-0934/$ – see front matter © 2004 Elsevier B.V. All rights reserved. doi:10.1016/j.jviromet.2004.08.007