APPLICATION OF DESIGNED MPCR PRIMER FOR E6 GENE OF HPV TYPE 45 AND 52 IN CERVICAL CANCER PATIENTS
Objective: Cervical cancer is the third disease that causes death in the world. The main cause of cervical cancer is Human papillomavirus (HPV). HPV have E6 and E7 oncogenes that, responsible for cancer incidence. One of molecular biology techniques for HPV identification is PCR. The aimed of this study is to applicate the multiplex PCR primer design for HPV type 45 and 52 identification.
Methods: DNA was isolated from 31 cervical cancer biopsy and cervical smear samples. DNA was amplicated with MY09/MY11 primer. Line probe array used to genotype of HPV. Application of primer design use MPCR methods with conditions was hot start 95 Â°C (3 min), denaturation 95 Â°C (15 s), annealing 58 Â°C (30 s), extension 72 Â°C (15 s) and final extension 72 Â°C (1 min). The amplified products were analyzed on a 1.5% agarose gel, stained with Gel Red and visualized in-gel documentation system.
Results: Result of HPV identification of HPV shown 19 of the 31 samples in this study was positive HPV. HPV genotyping was obtained the HPV type of the samples were HPV type 16, 18, 45 and 52. Application of design primer gives positive result. The band on S1, S2, S3 and S4 lane indicated the presence of gen E6 of HPV type 45 in J.1, J.3 and J.14 sample code and HPV type 52 in J.22 sample code.
Conclusion: MPCR primer design for E6 gene of HPV type 45 and 52 was applicated for identification of HPV type 45 and 52 in cervical cancer patients.
Keywords: Cervical cancer, Primer design, HPV type 45, HPV type 52, Multiplex PCR.
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