Molecular variants of human papilloma viruses type 16 and 6 in women with different cytological results detected by RFLP analysis [Detekcija molekularnih varijanti humanih papiloma virusa tipova 16 i 6 u žena s različitim citološkim promjenama RFLP analizom]

Žele-Starčević, Lidija and Plečko, Vanda and Skerlev, Mihael and Tripković, Vesna and Ljubojević, Suzana and Budimir, Ana and Mareković, Ivana and Lipozenčić, Jasna (2010) Molecular variants of human papilloma viruses type 16 and 6 in women with different cytological results detected by RFLP analysis [Detekcija molekularnih varijanti humanih papiloma virusa tipova 16 i 6 u žena s različitim citološkim promjenama RFLP analizom]. Collegium Antropologicum, 34 (3). pp. 887-891. ISSN 0350-6134

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Abstract

HPV infections are common and the presence of the same high-risk type in cervical specimens can be due to reinfection or persistence. Persistent infection is the most important predictor for development of cervical carcinoma. The aim of this study was to validate PCR-RFLP with two sets of primers: MY09/MY11 that amplify a fragment of L1 and P1/P2 that amplify a fragment of E1 ORF. PCR product of MY09/MY11 was digested with a set of 6 restriction enzymes (RE) and PCR product of P1/P2 with a set of 12 RE. Cervical samples from 110 women patients of the University Gynecologic Clinic CHC Zagreb were analyzed. There were 98 (89.1%) PCR positive samples detected with P1/P2 primers, and 94 (85.5%) PCR positive samples detected with MY09/MY11 primers. Seven HPV types were detected with P1/P2-RFLP technique and 17 with MY09/MY11-RFLP PCR positive samples amplified with both primer pairs agreed with each other in 82 samples; 16 samples were only positive with P1/P2 and 12 samples were only positive by MY09/MY11. HPV 16 was detected in 39 samples with MY09/11-RFLP, out of these two variants (two different patterns) were found with P1/P2 using Dde I, Hae III and Eco I. HPV 6 was detected in 9 samples with MY09/11-RFLP, out of these two variants were found with P1/P2 using HinfI. Combining these two PCR-RFLP methods subtypes of HPV 16 and HPV 6 were detected.

Abstract in Croatian

Infekcije uzrokovane humanim papiloma virusima (HPV) su česte te prisutnost istog genotipa visokog rizika u cervikalnim uzorcima može značiti reinfekciju ili perzistenciju. Perzistentna infekcija je najvažniji prediktor za razvoj karcinoma vrata maternice. Cilj ovog istraživanja je validacija dva seta primera: MY09/11 koji amplificiraju dio gena L1 i P1/P2 koji amplificiraju dio gena E1. Produkt amplifikacije s primerima MY09/11 izložen je digestiji sa 6 restrikcijskih enzima (RE), a produkt amplifikacije s P1/P2 s 12 RE. Analizirani su uzorci od 110 žena koje su se liječile u Klinici za ženske bolesti i porode KBC Zagreb. Amplifikacijom s P1/P2 dobiveno je 98 (89,1%) pozitivnih uzoraka, a s MY09/11 94 (85,5%). Sa MY09/11-RFLP tehnikom dobiveno je 17 različitih genotipova, a sa P1/P2-RFLP tehnikom 7. S oba para primera bilo je pozitivno 82 uzorka; 16 uzoraka bilo je pozitivno samo s P1/P2, a 12 samo s MY09/11. HPV 16 je nađen u 39 uzoraka tehnikom MY09/11-RFLP; od toga su P1/P2-RFLP tehnikom nađene 2 varijante (2 različita obrasca razgradnje) koristeći 3 RE (Dde I, Hae III and Eco I). HPV 6 je nađen u 9 uzoraka tehnikom MY09/11-RFLP; od toga su P1/P2-RFLP tehnikom nađene 2 varijante koristeći samo jedan RE (Hinf I). Kombinacijom 2 PCR-RFLP tehnike otkriveni su subtipovi HPV 16 i HPV 6.

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