Studi Pemetaan Awal DNA Mycobacterium tuberculosis Complex Secara Spoligotyping Pada Hasil Isolasi Dahak Pasien Tuberkulosis Paru Dari 10 Ibu Kota Propinsi (Bagian I)

Lisdawati, Vivi and Parwati, Dewi and Sudarmono, Pratiwi and Sudiro, T. Sudiro (2010) Studi Pemetaan Awal DNA Mycobacterium tuberculosis Complex Secara Spoligotyping Pada Hasil Isolasi Dahak Pasien Tuberkulosis Paru Dari 10 Ibu Kota Propinsi (Bagian I). Buletin Penelitian Kesehatan, 38 (4). ISSN 0125-9695

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Abstract

Mapping TB genotype of Mycobacterium tuberculosis (Mtb) is an important study to identify their distribution or characteristic and also may lead to improve control of the disease. This study conducted a preliminary mapping of the tubercle bacilli which had been circulating in Indonesia. Cultures of DNA isolated from TB patients at urban areas in 16 provinces in Indonesia, are chosen based on TB Case Detection Rate (CDR) 2006 from Indonesia Directorate General of Communicable Disease Control and Environment Health (Ministry of Health), were analyzed by spoligotyping for strain differentiation. In this first part, the analyzed result only came from urban areas in 10 provinces, i.e. Palembang, Bandar Lampung, Serang, Jakarta, Bandung, Surabaya, Banjarmasin, Makassar, Pontianak and Ambon. Sample were 269 DNA from 294 isolates collected from sputum of suspect TB patients with sputum-smear positive (SS+) and age above 15 years old. All samples were obtained from peripheral health laboratory in each province. The procedure collection is accordance to Indonesia DOTs guidelines (AFB smears) and samples were transported from those 10 areas to Bacteriology Laboratory at CBPRD. Sputum was taken for culture in liquid media MGIT Bactec 960 and solid media Lowenstein Jensen. The DNAs from positive liquid media MGIT Bactec 960 were isolated and analyzed by spoligotyping to identify the spoligo pattern. The spoligotyping results converted into octal format within Words & Excel spreadsheets and compared to International Spoligotype-database (SpolDB4). The previous study (Parwati et.al.) found some differences geographical distribution between Beijing genotype strain of tubercle bacilli in West Indonesia compare to East Indonesia, and the same pattern was also found in this study. Furthermore, the results in this study showed the differences in spoligo pattern of Mtb complex at 10 urban areas in West, Middle and East Indonesia. The percentage of Beijing strain family in the samples from West Indonesia showed around 26.61% (31.48% in Sumatra, 28.83% in Java and 16.98% in Kalimantan); from the Middle Indonesia around 25.93%; and none were found in samples from the East Indonesia. The SpolDB4 pattern also showed that the majority of isolates belonged to major clades of M.tuberculosis, i.e. the East African-Indian (EAI); Haarlem (H), Latin American-Mediterranean (LAM), the Central and Middle Eastern Asian (CAS); U = undefined; T = T family; and the MANU/ Manu family. We also found some isolates of Mycobacterium bovis. There were no significant association showed between genotype families and gender, but strong significant association found with ethnics and geography. Further confirmation of the results is still

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