PCR and genotyping for HPV in cervical cancer patients

PCR and genotyping for HPV in cervical cancer patients

Blog Article

Aims: To devise nested multiplex polymerase chain reaction (NMPCR) protocol for detection of mucosal human papilloma viruses (HPVs) and typing of HPV-16 and -18 in formalin-fixed, paraffin-embedded (FFPE) tissues of carcinoma cervix (CaCx).Settings and Design: Cross-sectional observational study.Materials and Methods: NMPCR was done for simultaneous detection of HPV, targeting 134 bp L1 capsid gene employing GP+/mGP+ primers and typing of genotypes-16 and -18, targeting E6/E7 gene from 34 FFPE tissue blocks of CaCx and cervical intraepithelial neoplasia (CIN).

Detection of 142 bp consensus sequence of L1 capsid gene was performed by nested PCR employing MY/GP+ primers.Sequencing of selected PCR amplicons of the later protocol obtained from control cell line DNA and 5 select samples were done for validation of the NMPCR luau thank you cards protocol.Statistical Analysis Used: Calculation of percentage from the Microsoft Excel Software.

Results: Of 26 FFPE samples of CaCx, 17 (65.3%) samples were found positive for HPV by NMPCR.Amplicons of 142 bp L1 capsid gene employing MY/GP+ primers were observed in 11 (42.

3%) samples of CaCx.Nearly 25% samples of CIN were positive for HPV.On sequence analysis, it was observed that the sample typed as HPV-16 by NMPCR was found to be the same on sequencing of amplicons obtained after MY/GP+ nested PCR.

Conclusions: This study indicates the usefulness of our NMPCR protocol for detection of mucosal HPVs and typing of HPV-16 and -18 from FFPE tissue samples of CaCx.The NMPCR protocol may be used to detect HPV and type common earthbath facial wipes genotypes-16 and -18 in fresh tissue of cervical biopsy or scrape samples for screening of CaCx.