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"Latar Belakang: Ada sekitar 50 spesies Nontuberculous mycobacteria (NTM) berpotensi patogen di manusia, yang menyebabkan infeksi tersering di paru. Pemeriksaan mikrobiologi berbasis molekuler diperlukan dalam mendiagnosis infeksi NTM paru. Oleh karena itu perlu dilakukan uji awal untuk deteksi mycobacteria dari spesimen klinis secara langsung dengan metode molekuler yaitu real-time PCR dan sekuensing DNA untuk identifikasi spesies mycobacteria serta mengaplikasikan pada kit PaxView® TB/NTM MPCR-ULFA. Tujuan: Melakukan optimasi berbasis molekuler untuk deteksi dan identifikasi Mycobacteria secara langsung pada sputum pasien terduga infeksi NTM paru. Metode: Studi dekskriptif dan eksperimental laboratorium dengan melakukan optimasi suhu penempelan, reaksi silang, ambang batas deteksi DNA, dan penerapan real-time PCR berbasis SYBR Green yang telahdioptimasi dan PaxView® TB/NTM MPCR-ULFA pada hasil sputum pasien terduga infeksi NTM paru. Hasil: Dua hasil positif dari 30 sampel sputum pada real-time PCR mycobacterium dan hasil sekuensing adalah Mycobacterium tuberculosis, menunjukkan adanya discordant hasil dengan real-time PCR MTB. Pada kit PaxView® TB/NTM MPCR-ULFA didapatkan 16 hasil positif MTB dan tidak ditemukan NTM. Kesimpulan: Terdapat discordance pada dua sampel hasil penerapan uji awal real-time PCR mycobacterium dengan sekuensing DNA, yang diduga NTM tetapi hasilnya M. tuberculosis. Perlunya dilakukan evaluasi lebih lanjut real-time PCR berbasis SYBR Green.

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Background: There are approximately fifty species of Nontuberculous mycobacteria (NTM) are potentially pathogenic in humans, the infection is most common in the lungs. Molecular-based microbiological examination is needed in diagnosing pulmonary NTM infection. Therefore, it is necessary to preliminary test the detection of mycobacteria from clinical specimens directly by molecular methods, namely real-time PCR and DNA sequencing to identify mycobacteria species and apply to the Pax-ULFA PaxView® TB/NTM kit. Aims: To perform Molecular-based optimization for the detection and identification of mycobacteria directly in sputum patient suspected of pulmonary NTM infection. Method: A descriptive and experimental laboratory study, to optimize the annealing temperature, determination of minimal detection of DNA, cross reaction of optimized real-time PCR based on SYBR- Green and applied sputum from patients suspected of NTM pulmonary infection to real-time PCR and PaxView® TB/NTM MPCR-ULFA. Results: Two positive results from 30 sputum samples on real-time PCR mycobacterium and sequencing results were MTB, the results discordant with real-time PCR MTB. In the PaxView® TB/NTM MPCR-ULFA, 16 positive MTB results were obatined and no NTM was found. Conclusion: There was discordance in two sample of real-time PCR mycobacterium spp. with DNA sequencing, which is thought to be NTM but the result is M. tuberculosis. The need for further evaluation of real-time PCR based Mikrobiologi Klinik on SYBR Green."

"Penyakit paru-paru akibat bakteri Nontuberculous Mycobacteria (NTM) semakin banyak terdeteksi pada kelompok rentan. Ketahanan NTM terhadap klorin mencapai 1000 kali lipat dibandingkan E. Coli, memungkinkan NTM bertahan di sistem distribusi air. Penelitian ini bertujuan menganalisis konsentrasi dan spesies NTM pada air bersih di rumah tangga dan Rumah Sakit X, serta mengestimasi dosis paparan NTM pada kelompok rentan menggunakan metode quantitative microbial risk assessment (QMRA). Metode penelitian meliputi enumerasi bakteri dengan media Middlebrook M7H10, elektroforesis, PCR, dan sanger sequencing. Rata-rata konsentrasi total NTM di rumah tangga Kecamatan Beji dan Sukmajaya, dengan sumber PDAM dan air tanah, adalah 14,7 CFU/L dan 11,3 CFU/L. Di Rumah Sakit X, konsentrasi rata-rata adalah 5,3 CFU/L. Ditemukan 2 spesies NTM pada 10 isolat (20%), yaitu Gordonia sp., Mycobacterium wolinskyi, dan Mycolicibacterium mucogenicum di rumah tangga, serta Streptomyces sp. di Rumah Sakit X. Estimasi dosis risiko paparan NTM menggunakan QMRA menunjukkan dosis paparan di rumah tangga sebesar 0,00434 CFU dan di Rumah Sakit X sebesar 0,00197 CFU. Meskipun konsentrasi NTM tergolong rendah, diperlukan langkah-langkah pengurangan paparan NTM seperti penggunaan filter dan UV, mengganti jenis perbandingan, serta mengurangi stagnasi air bersih untuk mengurangi risiko kesehatan.

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The incidence of lung Lung diseases caused by Nontuberculous Mycobacteria (NTM) are increasingly detected in vulnerable groups. NTM's resistance to chlorine is up to 1000 times greater than that of E. Coli, allowing NTM to persist in water distribution systems. This study aims to analyze the concentration and species of NTM in household and Hospital X clean water, and to estimate the NTM exposure dose to vulnerable groups using the quantitative microbial risk assessment (QMRA) method. The research methods included bacterial enumeration using Middlebrook M7H10 media, electrophoresis, PCR, and Sanger sequencing. The average total concentration of NTM in households in Beji and Sukmajaya Districts, with PDAM and groundwater sources, was 14.7 CFU/L and 11.3 CFU/L, respectively. At X Hospital, the average concentration was 5.3 CFU/L. Two NTM species were found in 10 isolates (20%), namely Gordonia sp., Mycobacterium wolinskyi, and Mycolicibacterium mucogenicum in households, as well as Streptomyces sp. in X Hospital. The NTM exposure risk dose estimation using QMRA indicated an exposure dose in households of 0.00434 CFU and 0.00197 CFU in Hospital X. Although the NTM concentration was relatively low, measures to reduce NTM exposure, such as using filters and UV, changing disinfectant types, and reducing clean water stagnation, are necessary to mitigate health risks."