Qualitative-based Multiplex PCR: Diagnosis of Papilloma virus Types 16 & 18 in Samples Taken from Patients with Malignant Lesions

Abstract Background and Aims: The human papilloma virus was introduced as the major etiological agent in outbreak of cervical cancer in 1970. Since it is very difficult to recognize these viruses and their types using the serological tests and cell culture, molecular methods such as PCR are of great importance. Therefore, in this study, our goal was to use a multiple specific PCR assay on L1 and E6 genes of HPV for molecular detection of this virus and its common type’s detection in the society. Materials and Methods: After collecting the samples from malignant lesions of various patients, the viral DNA was extracted from paraffin blocks of 50 clinical samples and the PCR method was performed on the mentioned samples by specific primers for L1 and E6 genes together with β-globin (as internal control). The PCR product was analyzed on 2℅ agarose gel and the sensitivity of this test was examined. Results: from among 50 samples of the patients, 33 cases were HPV positive and 17 ones were negative. The sensitivity of this test was 20 copies from recombinant construct containing target genes for each reaction. Conclusion: this study confirmed that the designed PCR with specific primers on L1 and E6 genes of HPV proved to be an accurate method for detecting and determining the HPV types.