Naslov
HPV Genotyping by PCR, RFLP, Sequencing and Line Probe Assay

Autori
Milutin, Nina ; Matovina, Mihaela ; Husnjak, Koraljka ; Grce, Magdalena

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Final Programme and Abstracts book of the 5th International Multidisciplinary Congress Eurogin 2003 / Monsonego, Joseph - Pariz : Eurogin, 2003

Skup
5th International Multidisciplinary Congress Eurogin 2003 "Preventing and Controlling Cervical Cancer in the New Millenium - Human Papillomavirus Infection and Neoplasia, a New Era - Sexually Transmitted Infections, the Global Picture"

Mjesto i datum
Pariz, Francuska, 13.04.2003. - 16.04.2003

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
Papillomavirus; Genotyping

Sažetak
Cervical cancer was in the third place following breast and stomach cancer in Croatia in 1993. HPV infection is a major risk factor for cervical cancer development. The objective of this study was to compare different methods of HPV genotyping: polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) analysis, sequencing and Line probe assay (LiPA, Innogenetics). The presence of HPV was evaluated in cervical samples collected at different gynaecological clinics from Zagreb (Croatia). HPV DNA was detected by PCR using two sets of universal primers (MY09/MY11 and L1C1/L1C2) and type-specific primers for HPV 6/11, 16, 18, 31 and 33 as previously described (1). A subset of MY09/MY11-positive high-grade neoplasia samples was analysed by RFLP and LiPA test. Undetermined HPV types by RFLP and/or LiPA were further analysed by sequencing. Out of 1669 specimens, 65% were positive, of which 61% were determined by types-specific directed-PCR and 38% remained untyped. The most frequently observed type was HPV 16 in 28% of HPV-positive samples. Among high-grade cervical intraepithelial neoplasia HPV-positive but untyped specimens 35 were chosen for further analysed by RFLP and LiPA of which some of them were also sequenced. One type of HPV was determined in 20 and 21 cases, multiple infections in 4 and 10 cases, by RFLP and LiPA, respectively. RFLP and LiPA did not determined HPV types in 11 and 4 cases, which were finally defined by sequencing. The advantage of sequencing and RFLP of PCR products over LiPA is to allow genotyping of more different HPV types, but the disadvantage is not to be able to discriminate multiple HPV infections. Thus, the advantage of LiPA over sequencing and RFLP is to be able to identify multiple HPV infections, which are obviously frequent (10/35) within high-grade neoplesia specimens shown herein.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

POVEZANOST RADA