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All Journal CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) INDONESIAN JOURNAL OF LEGAL AND FORENSIC SCIENCES Journal Pharmaceutical Science and Application (JPSA) Jurnal Kimia (Journal of Chemistry) METAMORFOSA Journal of Biological Sciences Buletin Udayana Mengabdi Jurnal Farmasi Udayana Jurnal Veteriner ANNALES BOGORIENSES JFIOnline
I Nengah Wirajana
Program Studi Kimia, Fakultas Sains Dan Matematika, Universitas Kristen Satya Wacana
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Dentistry Education Electrical & Electronics Engineering Environmental Science Health Professions Immunology & microbiology Law, Crime, Criminology & Criminal Justice Materials Science & Nanotechnology Medicine & Pharmacology Public Health Veterinary

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Identifikasi Mutasi Gen rpob Isolat MDR Mycobacterium tuberculosis di Bali dengan Metode Nested PCR*) Identification Of rpob Gene Mutation of MDR Mycobacterium tuberculosis Isolate in Bali Using Nested PCR Yowani, Sagung Chandra; Yowani, Sagung Chandra; Wirajana, I Nengah
JFIOnline | Print ISSN 1412-1107 | e-ISSN 2355-696X Vol 7, No 2 (2014)
Publisher : Indonesian Research Gateway

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Abstract

ABSTRAK Tuberkulosis merupakan salah satu penyakit infeksi yang menjadi perhatian untuk ditangani pada program Millenium Development Goals (MDGs), terutama oleh meningkatnya kejadian Multi Drug Resistance (MDR_TB). Menurut WHO, resistensi terhadap rifampisin dan isoniazid telah dikategorikan sebagai MDR-TB. Dari beberapa penelitian, ditemukan bahwa hampir 90 % isolat yang resisten rifampisin, juga resisten terhadap isoniazid. Oleh karena itu,resistensi terhadap rifampisin dianggap sebagai marka penentu (surrogate marker) kejadian MDR-TB. Resistensi terhadap rifampisin dimediasi oleh adanya mutasi pada suatu region 81 bp (RRDR : Rifampicin Resistance Determinant Region) dari gen rpoB. Penelitian ini bertujuan utamanya untuk mengeksplorasi mutasi pada gen rpoB dari MDR-TB yang terjadi pada penderita Tuberkulosis di Bali. Isolat MDR-TB yang digunakan pada penelitian ini merupakanj koleksi Laboratorium Mikrobiologi RS Sanglah, Denpasar. Amplifikasi dilakukan dengan metode Nested Polymerase Chain Reaction menggunakan sepasang outer primer dan sepasang inner primer Hasil penelitian menunjukkan selain mutasi pada daerah RRDR, 5 dari 6 isolat yang dikerjakan memiliki titik mutasi yang sama yaitu pada kodon 418, dengan perubahan asam amino dari asam glutamat menjadi asam aspartat.Oleh karenanya, dapat disimpulkan bahwa terdapat missense mutation   Kata Kunci : Nested PCR, MDR-TB, Mutasi gen rpoB ABSTRACT Tuberculosis is one of the  infectious diseases that is important to be treated in Millenium Development Goals (MDGs) programme, mainly due to the increasing of Multidrug resistant tuberculosis (MDR-TB) cases. According to WHO, MDR-TB was defined as tuberculosis caused by strains of Mycobacterium tuberculosis that are resistant to at least isoniazid and rifampicin. Several studies found that nearly 90% of rifampin-resistant isolates, were also resistant to isoniazid. Therefore, resistance to rifampicin is a surrogate marker of MDR. Resistance to rifampicin is mediated by  mutation in a region of 81 bp (RRDR: Rifampicin Resistance Determinant Region) of the rpoB gene. The aim of this research mainly was to explore mutation of MDR-TB’ rpoB gene of tuberculosis patient in Bali. All MDR-TB isolates used for this research were the collection of Clinical Microbiology Laboratory Of Sanglah Hospital, Denpasar. Amplification were conducted by Nested Polymerase Chain Reaction methods using two pair of  inner and outer primers. The result of this study showed that except at RRDR, five of six isolates had one similar mutation at codon 418 from glutamic acid to aspartic acid. Therefore, it could be concluded that there has been a missense mutation in all isolates. Key words: Nested PCR, MDR-TB, r.poB gene mutation
Co-Authors A. A. B. Putra Anak Agung Istri Agung Mayun Laksmiwati D. Rizkiyanti Darma Asih Yuliana Dewi Andayani Farmawati Dewi, I Gusti Ayu Agung Gangga Samala Diah Suci Eddy Bagus Wasito Gusti A Malelak H.M. Soekry Erfan Kusuma I Gede Mahardika I Gusti Ngurah Bagus Andre Hartawan I Ketut Gede Dharma Dewantara I Made Agus Gelgel Wirasuta I P. J. D. A. Suartama I Putu Mahendra I W BUDIARSA SUYASA I Wayan Gede Gunawan I Wayan Suarsa I. A. Gede Widihati Ida Ayu Ratih Dwi Nugraha Putri Ida Bagus Amertha Putra Manuaba Ida Bagus Putra Manuaba Ida Bagus Putu Eristya Putra Ika Kurniawati Indra Juana Adikara Inten Hardianti Nizar Iryanti Eka Suprihatin James Sibarani Kazuo Sakka Ketut Ratnayani Khisan Qamariya Kimura, Tetsuya Kusuma, Soekry Erfan Luh De Dwi Jayanthi Luk Ketut Budi Maitriani M. Manurung Made Arsa Made Dharmesti Wijaya Made Rai Dwitya Wiradiputra Mirawati N.K.W. N. M. T. Juliasari N. W. Bogoriani Ni Komang Ariati Ni Komang Lia Wahyuni Ni Luh Md. Widayantini Ni Luh Putu Mustika Praptiwi Ni Made Suaniti Ni Made Yustikarini Ni Nengah Kartini Asih Ni Nyoman Tri Puspaningsih Ni Putu Ariantari Ni Putu Citra Anggryni Sugitha Ni Putu Frida Oktaningtias Widiarthi Ni Putu Rahayu Artini O. Ratnayani Oka Ratnayani P. Suarya Pradnyaniti, D.G Putra, Pramana Kumala Putri, Ni Wayan Prasanthi Swarna Putu Elvira Yulianthi Putu Suarya R. R. Sirait Ro’yal Aini Sagun Chandra Yowani Sakka, Kazuo Satriya Putra Prakoso Soekry Erfan Kusuma Tetsuya Kimura Wahyu Dwijani Sulihingtyas Y. H. Pratiwi