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"Saat ini tidak ada metode standar untuk uji kepekaan obat terhadap Mycobacterium tuberculosis di antara laboratorium-laboratorium di Indonesia. Sejak bulan Januari 2002 sampai dengan Januari 2004, kami mencoba menerapkan metode plat dengan 25 sumur yang berisi media middlebrook’s (metode Drug Susceptibility Culture Plate (DSCP)) yang digunakan oleh Dutch Supranational Reference Laboratory at the Institute of Public Health and the Environment (RIVM), Bilthoven, Netherlands. Pengalaman kami memperlihatkan bahwa metode ini berpotensi memberikan hasil yang lebih baik karena sangat mudah distandardisasi, hasilnya lebih cepat dan dapat memperlihatkan nilai KHM (Konsentrasi Hambat Minimum) yang lebih terperinci. Data dari 364 isolat yang telah diuji dengan metode DSCP memperlihat resistensi terhadap INH, rifampisin, ethambutol, dan streptomisin secara berurutan adalah 21,4%; 19,8%; 15,7%; and 16,5%. Resistensi ganda didapatkan pada 13,2% isolat. (Med J Indones 2005; 14: 142-6)

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At present, there is no standardized method for Mycobacterium tuberculosis drug susceptibility testing (DST) among laboratories in Indonesia. Since January 2001 to January 2004 we have tried to establish the method of 25-well culture plates with middlebrook’s media (Drug Susceptibility Culture Plate (DSCP) method) used by the Dutch Supranational Reference Laboratory at the Institute of Public Health and the Environment (RIVM), Bilthoven, Netherlands. Our experience showed that this method potentially gives better result as it can be very well standardized, faster and provides detailed MIC (Minimal Inhibitory Concentration) values. Data from 364 isolates that have been tested by DSCP method showed that resistance to INH, rifampicin, ethambutol, and streptomycin were 21.4%, 19.8%, 15.7%, and 16.5% respectively. Multidrug resistance were found in 13.2% isolates. (Med J Indones 2005; 14: 142-6)"

"ABSTRAKRifampicin (RIF) adalah obat lini pertama untuk terapi tuberkulosis (TB) dengan kemampuan bakterisidal yang tinggi terhadap M. tuberculosis. Akan tetapi, kasus resistensi terhadap RIF telah menurunkan efektivitas terapi menggunakan obat ini. Multi-drug resistance TB (MDR/RR-TB) adalah tipe resistensi yang paling umum ditemukan pada galur MTB. Penggunaan obat berbasis peptida siklis telah banyak diminati karena peptida memiliki properti farmakologi yang baik, dan selektif. Studi ini bertujuan untuk menemukan kandidat senyawa peptida siklik yang potensial sebagai inhibitor protein RNA polimerase subunit β (RpoB) mutan S531L sebagai mutan dengan prevalensi tinggi pada MDR-TB. Struktur 3 dimensi dari RpoB wild type dan mutan S531L diunduh melalui basis data Protein Data Bank, dioptimisasi, dan dibandingkan karakteristik hidrofobisitas permukaannya untuk menentukan sekuens dari peptida siklis. Kemudian, basis data ligan peptida siklis dibuat dengan menggunakan generator kombinasi dan penggambaran. Optimisasi, kalkulasi muatan parsial dan minimisasi energi dilakukan pada basis data ligan, dan proses penapisan dilakukan dengan simulasi penambatan molekul. Simulasi ini dilakukan dengan dua tahapan rigid dan fleksibel terhadap struktur S531L RpoB, untuk menemukan ligan peptida siklis dengan interaksi paling baik dengan protein ini.  Simulasi ini menghasilkan 5 ligan terbaik dengan nilai energi bebas Gibbs terendah terhadap S531L RpoB. Ligan terpilih diprediksi sifat farmakologinya secara komputasi, dan menghasilkan 3 ligan (CYYEWC, CWYEGC, dan CQQNWC) yang memiliki karakter absorpsi, distribusi, metabolism, ekskresi, dan toksisitas yang sesuai. Ketiga ligan ini divalidasi interaksinya dengan menggunakan simulasi dinamika molekul, dan menunjukkan stabilitas interaksi yang baik sebagai kandidat obat untuk terapi MDR-TB.

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ABSTRACTRifampicin is the first line drug for tuberculosis (TB) treatment with high bactericidal activity towards M. tuberculosis. However, the rifampicin efficacy in TB treatment has been decreased steadily due to the emerging drug resistance cases. Among all types of the rifampicin resistance, MDR-TB is the most common resistance found in the MTB strain. Cyclic peptide therapeutics shows a significant success in the industry, since they possess a favorable pharmacological property. This study aimed to find the most potent cyclic peptide inhibitor for S531L RpoB protein for MDR/RR-TB treatment. Cyclic hexapeptide ligand database was built according to the binding site of the S531L RNA polymerase subunit β (RpoB) protein, the main mutation of the rifampicin target. The 3-dimension structure of RpoB wild type and mutant S531L were retrieved from the Protein Data Bank and optimized. Both of wild type and S531L binding site were compared based on their surface hydrophobicity to determine the cyclic peptide sequences in the ligand database. After the ligand database was built with combination generator and drawing, optimization, partial charge calculation and energy minimization were done. This database underwent two steps molecular docking simulation (rigid and flexible) against the S531L RpoB to find the cyclic peptide with best interaction towards this protein. The simulation resulted in 5 best ligands with the lowest value of Gibbs free energy binding to S531L RpoB. The selected ligands were subjected to the computational pharmacological properties prediction using several tools and resulted in three cyclic peptides (CYYEWC, CWYEGC and CQQNWC) with favorable interaction and ADME-Tox properties as MDR-TB drug candidate."

"Penyebaran Multidrug Resisten Tuberculosis (MDR TB) yang disebabkan oleh bakteri Mycobacterium tuberculosis merupakan perhatian untuk program penaoganan TB. Obat antituberkulosis lini kedua digunakan untuk pengobatan penderita MDR TB. Kami melakukao penelitian tentang Uji kepekaan obat antituberkulosis lini kedua menggunakan media Lowenstein Jensen dibandingkan dengan Mycobacterium Growth Indicator Tube (MGIT 960) sistem. Tiga puluh (30) isola!ba!cteri MDR TB di uji dengan oflokasin, amikasin, dan kanamisin menggunakan MGIT 960 dan baslinya dibandingkan dengan metode proporsi pada media Lowenstein Jensen. Dati basil penelitian didapat 27 isolat (90 %) sensitif teihadap ofloksasin , 21 isolat (70 %) sensitif terbadap antikasin dan 26 isolat (86,6 %) sensitif terhadap kanamisin. Dua isolat merupakan Extensively Drugs Resistance (XDR TB). Waktu untuk uji kepekaan dengan MGIT adalab 9 hari sedaogkan dengan metode proporsi 21 hari.

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The emergence of multidrug resistant tuberculosis (MDR TB) caused by Mycobacterium tuberculosis is real threat for TB control program. Second line drogs was using for person who has MDR TB. The objective of this study was to evaluate the proportion method for testing of Mycobacterium tuberculosis susceptibility to second line drugs compared to the Mycobacterium Growth Indicator Tube (MGJT 960 )System. Thirty MDR TB Isolates were tested for susceptibility to ojloxacin, amikasicin, and kanamycin by MGJT 960, and the result were compared to those obtain with proportion method on Lowenstein Jensen media, considered a reference method. Result for ojloxacin were 27 isolate (90 %) sensitive,21 isolate (70 %) sensitive to amikacin and 26 isolate (86,6 %) sensitive to kanamycin. Two Isolate were Extensively Drug resistance (XDR TB)The time required to obtain result was an average of 9 days by the MGIT and 21days by the reference method."

"Penyakit TB masih merupakan masalah kesehatan kesehatan masyarakat yang penting di Indonesia. Upaya diagnostik TB paru masih terus ditingkatkan. Pemeriksaan penunjang diagnosis TB yang sekarang digunakan masih mempunyai sensitiviti dan spesitiviti yang rendah. Tujuan penelitian mengetahui tingkat akurasi uji tuberkulin dan PCR terhadap penegakkan diagnosis TB serta hubungan uji tuberkulin dan PCR dengan BTA mikroskopis dan biakan M. Tb dalam diagnosis TB paru.Metode penelitian cross-sectional, uji diagnostik dan analisa data menggunakan Chi-Square. Kriteria inklusi penderita terdapat gejala klinik riwayat batuk 3 minggu disertai atau tanpa batuk darah, nyeri dada, sesak napas dan riwayat minum obat TB dalam jangka waktu kurang dari 1 bulan serta bukan TB (kontrol). Seluruh sampel dilakukan anamnesis, pemeriksaan fisik, lekosit, LEDI/II, foto toraks, uji tuberkulin, PCR, BTA mikroskopis 3X dan biakan M. Tb mctode kudoh. Baku emas yang digunakan biakan M. Tb metode kudoh. Data diolah menggunakan SPSS versi 11.00.Berdasar 127 sampel masuk kriteria inklusi 121. Sampel berjumlah 121 terdiri dari 61 sampel tersangka TB dan 60 sampel kontrol Sensitiviti dan spesivisiti uji tuberkulin terhadap biakakn metode Kudah menggunakan cut-off point 15,8 mm 33% dan 93%. Sensitiviti PCR terhadap biakab metode Kudoh 100%, spesitiviti PCR 78%. Didapatkan perbedaan bermakna dan hubungan lemah uji tuberkulin dengan biakan M. Tb dan PCR serta didapatkan perbedaan dan hubungan bermakna PCR dengan BTA mikroskopis biakan M. Tb.Kesimpulan basil keseluruhan penelitian mendapatkan basil 39 sampel biakan positif, 36 sampel BTA mikroskopis positif, 57 sampel PCR positif dan 18 sampel uji tuberkulin positif. Ditemukan sensitiviti basil uji tuberkulin lebih rendah daripada PCR, BTA mikroskopis dan biakan M. Tb mctode Kudoh. Meskipun terdapat perbedaan bermakna basil uji tuberkulin pada biakan positif clan negatif, BTA mikroskopis positif dan negatif, serta PCR positif dan negatif, akan tetapi uji tuberkulin (menggunakan cut-off point 15.8 mm) kurang dapat membantu penegakan diagnosis TB para. Berdasarkan hasil penelitian ditemukan bahwa diantara keempat pemeriksaan penunjang diagnosis TB paru PCR mempunyai nilai sensitivit dan spesitiviti tinggi ( 100% dan 78%). sehingga PCR dapat digunakan sebagai pemeriksaan penunjang diagnosis TB paru apabila didapatkan klinis dan radiology mendukung TB paru. Menggunkan pemeriksaan PCR akan didapatkan metode penegakan diagnosis TB paru yang cepat ( 1 hari ) dibandingkan dengan menunggu hasil biakan M. Tb hingga 8 minggu.

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Objective. In an attempt diagnosis pulmonary tuberculosis still increased continuously. Now additional examination pulmonary tuberculosis have been lack sufficient sensitivity and sensitivities. The aim of this study was to determine the validity of tuberculin skin testing (TST) and PCR toward assessment diagnosis pulmonary of tuberculosis with correlation between tuberculin skin testing to PCR with AFB microscopic and solid media culture of M. tuberculosis for the diagnosis of pulmonary tuberculosis.

Method. A cross-sectional study, diagnostic test and analysis with Chi-Square test. Inclusion criteria patient with pulmonary symptom include chronic cough 3 weeks with or without hemoptysis, chest pain, breathlessness and past history of ATA less than 1 month with non-tuberculosis patient (control). The general samples was examination Ro thorax, tuberculin skin testing, PCR, AFB microscopic and conventional culture. The golden standard is conventional culture test using Kudoh method. Analyze of the data with SPSS version 11.0.

Result. The study material comprised 121 samples from 127 samples. These samples include 61 samples from patient with probably active pulmonary tuberculosis and 60 control comprising healthy individuals. The sensitivity and specificity of tuberculin skin testing with cut-off point 15.8 mm greater was 33% and 93% on conventional culture test using Kudoh method. PCR sensitivity was 100% and spesitivity was 78%. It was showed the positivity correlation between pulmonary tuberculosis and conventional culture as well as PCR and AFB microscopic, the conventional culture test.

Conclusion. The sensitivity of tuberculin skin testing less than PCR, AFB microscopic and conventional culture test. So that not enough to assessment diagnosis pulmonary tuberculosis. The sensitivity and specificity PCR was I00% and 78%. With the use of PCR test, we were able to detect diagnosis pulmonary tuberculosis more rapidly in less than I day, compared to average 8 week required for detection by conventional culture."

"ABSTRAK Latar Belakang: dahak BTA mikroskopik adalah teknik diagnostik cepat yang palingbanyak digunakan untuk mendiagnosis TB paru. Di RSUP Persahabatan, pemeriksaandahak BTA mikroskopik membutuhkan tiga pemeriksaan sampel dahak lebih dari duahari, (sewaktu pertama, pagi, sewaktu kedua). Pengambilan dua sampel dahak akanmengurangi waktu, uang dan beban kerja laboratorium mikrobiologiTujuan: Penelitian ini bertujuan untuk mendapatkan sensitivitas dan spesifisitaspemeriksaan dahak BTA mikroskopik dari tiga bahan pemeriksaan dahak dan duabahan pemeriksaan dahak.Metode: Kami secara acak mengumpulkan data demografi dan hasil laboratorium (hasildahak BTA dan hasil biakan M.tuberculosis) dari pasien terduga TB kasus baru di RSUPPersahabatan dari data bulan Januari 2012 hingga Desember 2013. Subjek harus berusiaminimal 15 tahun dan bukan pasien TB MDR dan bukan pasien HIVHasil: Dari 360 pasien terduga TB paru, sebanyak 210/360 (58,3%) memiliki hasilbiakan M. tuberculosis positif. Sensitivitas 100% dan spesifisitas 69.2% pada hasilpemeriksaan BTA mikroskopik tiga kali (S1, P, S2) dan hasil pemeriksaan BTAmikroskopik dua kali sensitivitasnya 89.8% dan spesifisitas 94.1%. Kesimpulan: Hasil pemeriksaan dahak BTA mikroskopik 2 kali pemeriksaan memberikan spesifisitas yang lebih tinggi bila dibandingkan dengan hasil pemeriksaan3 kali dahak BTA mikroskopik dan memiliki sensitivitas yang masih cukup tinggi.ABSTRACT Background: Sputum smear microscopy is the rapid diagnostic technique that mostwidely used for diagnosing pulmonary tuberculosis. In Persahabatan hospital, sputumsmear microscopy requires three sputum sample examinations over two days, (spot1,morning, spot2). Collection of two sputum samples would reduce time, money andwork load of microbiology laboratory.Objectives: This study aimed to get sensitivity and specificity of sputum smearmicroscopy from three sputum samples collection and two sputum samples collection.Methods: We randomly collected demography and laboratory data (sputum smear andM.tuberculosis culture result) of presumptive TB new cases in Persahabatan hospitalfrom January 2012 to Desember 2013. Subjects must be 15 years old minimum, nonmultidrug-resistant TB presumptive and have non reactive HIV status.Results: Of 360 presumptive TB patients, 58.3% have M.tuberculosis positive cultureresults. Senstivity and specificity of sputum smear microscopy from three sputumsamples collection (S1, M, S2) are 100% and 69.2%. Smear microscopy of two samplescollection Sensitivity and specificity sputum smear microscopy 89.8% and 94.1%. Conclusion: Sputum smear microscopy test result from 2 sputum collections gives increase specificity compared to smear microscopy of 3 samples and have highly sensitivity.;Background: Sputum smear microscopy is the rapid diagnostic technique that mostwidely used for diagnosing pulmonary tuberculosis. In Persahabatan hospital, sputumsmear microscopy requires three sputum sample examinations over two days, (spot1,morning, spot2). Collection of two sputum samples would reduce time, money andwork load of microbiology laboratory.Objectives: This study aimed to get sensitivity and specificity of sputum smearmicroscopy from three sputum samples collection and two sputum samples collection.Methods: We randomly collected demography and laboratory data (sputum smear andM.tuberculosis culture result) of presumptive TB new cases in Persahabatan hospitalfrom January 2012 to Desember 2013. Subjects must be 15 years old minimum, nonmultidrug-resistant TB presumptive and have non reactive HIV status.Results: Of 360 presumptive TB patients, 58.3% have M.tuberculosis positive cultureresults. Senstivity and specificity of sputum smear microscopy from three sputumsamples collection (S1, M, S2) are 100% and 69.2%. Smear microscopy of two samplescollection Sensitivity and specificity sputum smear microscopy 89.8% and 94.1%. Conclusion: Sputum smear microscopy test result from 2 sputum collections gives increase specificity compared to smear microscopy of 3 samples and have highly sensitivity.;Background: Sputum smear microscopy is the rapid diagnostic technique that mostwidely used for diagnosing pulmonary tuberculosis. In Persahabatan hospital, sputumsmear microscopy requires three sputum sample examinations over two days, (spot1,morning, spot2). Collection of two sputum samples would reduce time, money andwork load of microbiology laboratory.Objectives: This study aimed to get sensitivity and specificity of sputum smearmicroscopy from three sputum samples collection and two sputum samples collection.Methods: We randomly collected demography and laboratory data (sputum smear andM.tuberculosis culture result) of presumptive TB new cases in Persahabatan hospitalfrom January 2012 to Desember 2013. Subjects must be 15 years old minimum, nonmultidrug-resistant TB presumptive and have non reactive HIV status.Results: Of 360 presumptive TB patients, 58.3% have M.tuberculosis positive cultureresults. Senstivity and specificity of sputum smear microscopy from three sputumsamples collection (S1, M, S2) are 100% and 69.2%. Smear microscopy of two samplescollection Sensitivity and specificity sputum smear microscopy 89.8% and 94.1%. Conclusion: Sputum smear microscopy test result from 2 sputum collections gives increase specificity compared to smear microscopy of 3 samples and have highly sensitivity."

"Tuberkulosis (TBC) masih menjadi penyebab utama kematian akibat penyakit menular oleh adanya infeksi. Rifampisin dan isoniazid adalah obat lini pertama yang paling efektif melawan infeksi Mycobacterium tuberculosis. Deteksi resistansi OAT yang tepat, akurat, dan komprehensif, serta pemilihan sampel diperlukan untuk memastikan diagnosis penyakit tuberkulosis pasien. Penelitian ini bertujuan untuk menganalisis perbandingan hasil targeted drug sequencing dari hasil dekontaminasi sputum dengan isolat Mycobacterium tuberculosis dan mengetahui kesesuaian DST fenotipik MGIT, genotipik GeneXpert dalam mendeteksi resistansi rifampisin dan isoniazid. Sampel penelitian ini adalah sampel sputum yang sudah ada hasil GeneXpert positif dan isolate kultur dengan hasil DST MGIT. Hasil dekontaminasi sputum langsung dan kultur positif dari sampel yang sama dilakukan targeted drug sequencing dengan Oxford Nanopore technology menggunakan flowcell MinION Mk1B. Hasil penelitian menunjukkan bahwa pada target gen rpoB pada 5 dari 6 sampel isolat kultur memberikan hasil gen resistan rpoB dan 1 undetermined. Pada sebagian besar dekontaminasi sputum yaitu 5 dari 6 sampel juga memberikan hasil resistan terhadap rpoB dan 1 dekontaminasi sputum yang undetermined. Hasil resistansi obat isoniazid didapatkan pada target gen inhA sebanyak 5 dari 6 isolat kultur memberikan hasil sensitif pada inhA dan 1 isolat undetermined. Sedangkan pada dekontaminasi sputum 4 dari 6 sampel memberikan hasil sensitif pada inhA dan 2 undetermined. Lalu, pada target gen katG terdapat 3 dari 6 isolat kultur memberikan hasil sensitif, 2 isolat resistan, dan 1 undetermined. Sedangkan pada dekontaminasi sputum memberikan 2 hasil sensitif, 2 hasil resistan, dan 2 hasil undetermined. Metode targeted drug sequencing dapat dilakukan dari sampel hasil dekontaminasi sputum dan isolat. Keberhasilan banyak didapatkan dari hasil kultur dibandingkan dekontaminasi sputum. Pemeriksaan dengan targeted drug sequencing memberikan hasil yang sesuai dengan hasil DST MGIT dan GeneXpert untuk deteksi gen resisten Rifampisin (rpoB) dan Isoniazid (inhA dan katG).

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Tuberculosis (TBC) is still the main cause of death due to infectious diseases. Rifampicin and isoniazid are the most effective first-line drugs against Mycobacterium tuberculosis infection. Precise, accurate and comprehensive detection of OAT resistance, as well as sample selection are needed to confirm the patient's diagnosis of tuberculosis. This study aims to compare the results of targeted drug sequencing from sputum decontamination results with Mycobacterium tuberculosis isolates and determine the suitability of MGIT phenotypic and GeneXpert genotypic DST in detecting rifampicin and isoniazid resistance. The samples for this study were sputum samples that had positive GeneXpert results and culture isolates with DST MGIT results. The results of direct sputum decontamination and positive culture from the same sample were subjected to targeted drug sequencing with Oxford Nanopore technology using a MinION Mk1B flowcell. The results showed that for the rpoB gene target, the majority of culture isolates from 5 of the 6 culture isolate samples gave rpoB resistance gene results and 1 was undetermined. In the majority of sputum decontamination, 5 out of 6 samples also gave resistance to rpoB and 1 sputum decontamination was undetermined. Isoniazid drug resistance results were obtained for the inhA gene target, 5 of the 6 culture isolates gave sensitive results for inhA and 1 isolate was undetermined. Meanwhile, in sputum decontamination, 4 of the 6 samples gave sensitive results for inhA and 2 were undetermined. Then, for the katG gene target, 3 of the 6 culture isolates gave sensitive results, 2 isolates were resistant, and 1 was undetermined. Meanwhile, sputum decontamination gave 2 sensitive results, 2 resistant results, and 2 undetermined results. The targeted drug sequencing method can be carried out from samples resulting from decontamination of sputum and isolates. Much success comes from culture results rather than sputum decontamination. Examination with targeted drug sequencing provided results that were in accordance with the results of DST MGIT and GeneXpert for the detection of Rifampicin (rpoB) and Isoniazid (inhA, and katG) resistance genes."

"ABSTRAKTeknologi terus dikembangkan dalam dunia biomedis untuk menemukan cara pengobatan yang tepat dan murah. Sebuah metode untuk mengukur efekifitas anti mikroba adalah dengan metode uji silinder. saat ini, pengukuran zona inhibisi masih dilakukan dengan menggunakan caliper atau penggaris. Dalam penelitian ini, sistem instrumentasi untuk deteksi otomatis dan pengukuran zona inhibisi dikembangkan dengan menggunakan teknik pengolahan citra. Sampel zona inhibisi dalam cawan petri ditangkap menggunakan kamera dan diproses secara langsung di komputer.Pada penelitian ini perangkat lunak dibuat dengan memanfaatkan program Matlab berbasis Graphical User Interface GUI , Akhirnya, sistem pengukuran yang diusulkan sangat menjanjikan dan dapat digunakan sebagai alat untuk evaluasi efektivitas anti mikroba.

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ABSTRACTTechnology continues to be developed in the biomedical world to find the rightand inexpensive treatment. A method for measuring anti microbial effectiveness is by cup platemethod. Currently, inhibition zone measurements are still done manually using a caliper or ruler.In this study, the intrumentation sistem for automatic detection and zone measurement of inhibitionwas developed using image processing techniques. Samples of inhibition zones in petri dishes werecaptured using cameras and processed directly on the computer. In this research the software ismade by using Matlab program based on Graphical User Interface GUI . Finally, the proposedmeasurement sistem is very promising and can be used as a tool for evaluation of anti microbialeffectiveness. "