Detection of human papilloma virus (HPV) in parafin block of cervical cancer patients using multiplex polymerase chain reaction (PCR) and reverse line blot methods

Abstract

ObjectivesThe aim of this study is to detect human papilloma virus (HPV) especially types 16 and 18 through DNA expression from paraffin block preparations of cervical cancer and pre-cancer. MethodsThis study was conducted at the Anatomical Pathology Laboratory and Molecular biology Hasanuddin University Medical Research Center (HUMRC) Laboratory. The samples that used were came from cervical specimens which had been processed into paraffin blocks and had been diagnosed as Cervical Intra-epithelial Neoplasia (CIN) and Cervical carcinoma. HPV DNA was extracted from paraffin blocks using DNA extraction reagents from FFPE (Qiagen). DNA extraction products were then propagated by the polymerase chain reaction (PCR) method. HPV genotyping is done by a reverse line blot method using Ampliquality HPV-Type Express (AB Analitica). ResultsTotally 67 cervical samples from paraffin blocks preparations were diagnosed in Squamous Cell Carcinoma (SCC) and Cervical Intra-epithelial Neoplasia (CIN). The large number of cancer patients was more than 40 years old (42 patients/67.7%), with most of which are in the periods 41–50 years old (25 patients/40.3%). The most common cancer type was Non Keratinizing Squamous Cell Carcinoma (NK-SCC) with 50 (74.6%) samples. The most infectious strains are HPV strains 16, which were found in 53 (79.1%) samples both in single infection and co-infection with other HPV strains. Three of four HPV strains found were 16, 18 and 52 strains that belonging to High Risk strain of HPV, while strain 67 was included in the low risk group. There was also co-infection of 2 different HPV strains which involved 16 & 67 HPV Co-infection and 52 & 67 HPV Co-infection. ConclusionCervical cancer patients were more commonly found in women with more than 30 years old, which the most common type was Non keratinizing squamous cell carcinoma (NK-SCC). HPV infection is strongly associated with cervical cancer. HPV 16 infection was found in many cervical cancer lesions both as a single infection and co-infection with other types of low risk HPV.