Naslov
Evaluation Of Consensus Primer Sensitivity For Human Papillomavirus (HPV) Detection By Polymerase Chain Reaction (PCR)

Autori
Sabol, Ivan ; Golubić Talić, Jasminka ; Grce, Magdalena

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

Izvornik
Ninth International Summer School on Biophysics - Book of Abstracts / Pifat-Mrzljak, Greta ; Ilakovac Kveder, Marina - Zagreb : Institut Ruđer Bošković, 2006

Skup
Ninth International Summer School on Biophysics - Supramolecular Structure and Function

Mjesto i datum
Rovinj, Hrvatska, 16.09.2006. - 28.09.2006

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
HPV

Sažetak
Human papillomaviruses (HPVs) belong to the family Papovaviridae. HPVs are strictly species specific. They infect epithelial cells either of the skin or the anogenital and oropharyngeal mucosa. Until now, 130 HPV types have been identified and fully sequenced. Approximately 40 HPV types infect the anogenital tract, of which 15 to 18 high risk types are commonly found in anogenital cancer biopsy specimens, notably cervical cancer. Detection and typing is useful and important for the diagnosis of HPV associated diseases, especially cervical precancerous lesions and cervical cancer. The molecular methods used for HPV testing are based on the method of hybridization, DNA amplification (polymerase chain reaction – PCR) or both. The PCR method is, actually, the most specific and the most sensitive for revealing the presence of otherwise undetectable quantities of HPV DNA. The method allows detection of wide spectrum of HPVs by using consensus (general) primers. The identification of HPV types may be performed either by hybridization with type-specific probes or by PCR using type-specific primers. The aim of this study was to evaluate the sensitivity of HPV detection by PCR using different consensus primers, i.e. MY09/11, PGMY09/11, L1C1/L1C2 and GP5+/6+ concensus primers. DNA isolated from HeLa cell line (derived from human adenocarcinoma, containing HPV 18 DNA), SiHa cell line (derived from cervical squamous cell carcinoma, containing HPV 16 DNA) and pBR322 plasmid DNA containing HPV types 16, 18 and 33 and pUC19 plasmid DNA containing HPV type 6 were used as positive controls, while A431 cell line DNA (derived from human epidermoid carcinoma) was used as negative control. The amplification by each set of primers was performed on different quantities (2000, 500, 20, 10 and 2 pg/ul) of HeLa or SiHA DNA diluted by A431 DNA in a final concentration of 40, 20, 10 or 2 ng/ul. Futhermore, different concentration (30, 3, 0.3 and 0.03 fg/ul) of HPV cloned DNA dissolved in 2 ng/ul of A431 DNA was also tested by each set of primers. The evaluation of each primers set will be presented.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

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