Sekuens Gen Protein Kapsid Mayor L1 Human Papilomavirus 16 dari Isolat Klinik Asal Bandung

Anandayu Pradita, Edhyana Sahiratmadja, Sony Suhandono, Herman Susanto

Abstract


Kanker serviks disebabkan oleh infeksi kronik human papillomavirus (HPV) dengan genotipe HPV-16 sebagai HPV tersering yang menginfeksi epitel serviks. Protein penyelubung virus yang disebut kapsid mayor (L1) mempunyai peranan penting dalam menginfeksi epitel serviks. Tujuan penelitian untuk mengisolasi dan menganalisis sekuens gen L1 HPV-16. Pengetahuan mengenai sekuens gen L1 dapat memberikan informasi yang berguna, salah satunya yaitu untuk pengembangan vaksin. Pada studi ini, deoxyribonucleic acid (DNA) virus diekstraksi dari sediaan biopsi pasien kanker serviks yang diambil pada bulan Juni sampai Oktober 2010 di Kebidanan dan Kandungan RS Dr. Hasan Sadikin Bandung. Gen diamplifikasi dengan polymerase chain reaction menggunakan primer spesifik. Infeksi HPV-16 pada jaringan kanker dikonfirmasi dengan menggunakan kit komersial untuk tes genotipe HPV. Fragmen L1 kemudian diklon dan diinsersikan ke dalam pJET1.2/L1-16, kemudian dipotong dengan enzim BamHI dan BgIII untuk kemudian divalidasi dan disekuensing. Hasil sekuensing menunjukkan amplikon gen L1 HPV-16 sebesar 1.595 pasang basa. Analisis dari dua amplikon gen L1 HPV-16 menggunakan software BIOEDIT dan Basic Local Alignment Search Tool menunjukkan kesamaan ho mologi 99% dan 97% dengan sekuens L1 HPV-16 asal Thailand yang terregistrasi pada GenBank. Simpulan, telah dilakukan kloning sekuens gen L1 HPV-16 dari dua isolat klinik Bandung. Hasil kloning HPV-16 pada penelitian ini memberikan informasi tentang variasi sekuens yang perlu dipertimbangkan bagi pengembangan vaksin terutama bagi daerah spesifik seperti penduduk asal Indonesia.

Kata kunci: Human papillomavirus, kanker serviks, gen L1 HPV-16
 

Sequence of Human Papilomavirus 16 Major Capsid L1 Gene from Clinical Isolates in Bandung

Cervical cancer is strongly associated with chronic human papillomavirus (HPV) infection. HPV-16 is the most prevalent genotype infecting cervical epithelium. The major coat protein of viral particle (L1) plays a key role in the infection process. Our study aimed to isolate the HPV-16 L1 gene and analyze its sequence. Samples used were samples collected from the Department of Obstetrics and Gynaecology, Dr. Hasan Sadikin General Hospital, Bandung during the period of June to October 2010. In this study, the HPV-16 L1 sequence was analyzed from the viral deoxyribonucleic acid (DNA) extracted from biopsy sample of cervical cancer patient biopsy samples.The HPV-16 L1 amplification was performed using the polymerase chain reaction with specific primer. The HPV infection in the cervical tissue was confirmed by commercial HPV genotyping test. The L1 fragment was cloned into plasmid and the insert of the recombinant clone pJET1.2/L1-16 was digested using BamHI and BgIII. The amplicon result showed HPV-16 L1 gene with a length of 1.595 base pairs. The sequence analysis of two samples using software BIOEDIT dan Basic Local Alignment Search Tool revealed a high level of sequence similarity to L1 HPV-16 from Thailand (99% and 97%) as registered in GenBank. In conclusion, the L1 HPV-16 gene from Bandung isolates revealed variations from published sequence. Knowledge on L1 gene sequence may give additional information to the development of vaccine. Further study on vaccine development is currently ongoing using this HPV-16 clone that may be specific to Indonesian population.

Key words: Cervical cancer, human papillomavirus, L1 HPV-16

 

DOI: 10.15395/mkb.v46n3.317


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