Cervical cross sectional study will be carried out

Cervical intraepithelial neoplasia (CIN) is an important public health problem. It is the leading cancer in Indian women and second most common malignancy among women worldwide 1. The major cause of CIN is chronic infection of cervix with the sexually transmitted Human papillomavirus 2. To date more than 150 HPV genotypes have been characterized, and about 40 of them infect the genital tract 3. The current estimates indicate approximately 1, 32,000 new cases of cervical cancer which are diagnosed and 74,000 deaths annually in India 4.Although new HPV infections are most common in young sexually active women, women aged over 25 years remain at risk of HPV infection. HPV rarely causes symptoms in early stages and so many women go undiagnosed 5. Indian women usually report late to the G-OPD and at an advanced stage. CIN can be diagnosed by biopsy and colposcopy hence, this screening of HPV 6 and its genotype in cervical lesion can provide useful information about the malignant potential of precursor lesion of cervical carcinoma and also a relation between CIN and HPV.It will also benefit these patients in the early detection and timely treatment of CIN.Objectives:• To screen for cervical intraepithelial neoplasia.• To screen for cervical lesion through histopathology.  • To evaluate the HPV infection in cervical lesion using PCR method.Methodology:A prospective and cross sectional study will be carried out for 2 months  on 30 women (aged 17-50yrs) who will be examined in the gynecology OPD of a tertiary care hospital, their medical history will be taken. Pap smear will be collected and colposcopy will be done. Those positive by Reid Index on colposcopy and willing for biopsy will be taken for biopsy in OT under anesthesia after informing them and obtaining a written consent.Biopsy will be taken from the aceto- white areas of cervix, will be stored and will be sent for histopathology at the department for the HPV screening. Following methods will be used:DNA ExtractionAreas selected by the reviewing pathologist will be used to isolate the DNA. Deparaffinization will be done with xylene and Proteinase K lysis will be done to extract the Genomic DNA form the cells. The isolated DNA will be resuspended in water and store at 4°C. Quantification of DNA will be done by UV Spectrophotometry.Agarose Gel Electrophoresis                                                                                                                   Gel electrophoresis of the genomic DNA will be carried out for qualitative analysis of samples prepared. A good DNA preparation appears as a sharp single band. A submarine horizontal agarose slab gel apparatus. 7 HPV GenotypingViral genotyping will be performed using the commercial kit of HPV Genotyping with positive control. Amplification reactions for PCR analysis were carried out as per the standardized protocol with a final volume of PCR reaction mixture at 25 ?l. 8  Implications:Our main aim in this study will be to screen for HPV in cervical lesions of women attending G-OPD. Our study will help in screening major HPV genotypes which causes CIN. This study will help the patient in many ways as it will help in early detection and timely treatment of the HPV infection which is asymptomatic and is the major cause for CIN. It will also help in collection of data regarding which age groups have more chances of getting infected from HPV