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All Journal Majalah Kedokteran Bandung Althea Medical Journal The Indonesian Biomedical Journal Sari Pediatri Jurnal Respirologi Indonesia
Lidya Chaidir
Department Of Biomedical Sciences, Faculty Of Medicine Universitas Padjadjaran
Biochemistry, Genetics & Molecular Biology Health Professions Medicine & Pharmacology Public Health

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Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis Linda Choerunnisa; Coriejati Rita; Anna Tjandrawati; Lidya Chaidir; Ida Parwati
Majalah Kedokteran Bandung Vol 52, No 4 (2020)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15395/mkb.v52n4.2141

Abstract

Indonesia is one of the countries with the highest multidrug-resistant tuberculosis cases in the world. Rapid molecular test using the Xpert MTB/RIF assay is one of the detection methods for MDR-TB. Early detection of MDR-TB is crucial for early initiation of treatment. However, Xpert MTB/RIF assay only detects the rpoB gene mutations associated with Rifampicin resistance. Recently, WHO recommends the use of Pyrosequencing, a DNA sequencing method that can detect not only the rpoB gene but also katG and/or inhA gene mutations associated with Isoniazid resistance. The aims of this study were to compare the interpretation between the two methods  and to determine the differences in codon mutation position detection of the rpoB gene and mutation detection of the katG and/or inhA gene. This was a cross-sectional comparative observational study on patients ≥18 years old interpreted as RR-TB patients based on Xpert MTB/RIF assay results who had not received MDR-TB drugs at Dr. Hasan Sadikin General Hospital, Bandung, Indonesia. Results showed there were 40 Rifampicin-resistant TB subjects interpreted by Xpert MTB/RIF assay while Pyrosequencing interpreted 30 MDR-TB, 9 RR-TB and one Isoniazid-resistant TB subjects in January - February 2020. The detection of rpoB gene codon mutation position between Xpert MTB/RIF assay and Pyrosequencing methods was not significantly different (p=0.389). Pyrosequencing had detected 27 katG gene mutations, 3 inhA gene mutations, one katG and inhA gene mutation. To conclude, Pyrosequencing can be used for accurate detection of Rifampicin and Isoniazid resistance in MDR-TB. Perbandingan Hasil Interpretasi antara Pyrosequencing dengan Xpert MTB/RIF Assay pada Multidrug-Resistant TuberculosisIndonesia merupakan salah satu negara dengan kasus multidrug-resistant tuberculosis terbanyak. Penegakan diagnosis MDR-TB saat ini menggunakan tes cepat molekular Xpert MTB/RIF assay sehingga pasien segera mendapatkan pengobatan. Namun Xpert MTB/RIF assay hanya mendeteksi mutasi gen rpoB penyandi resistansi Rifampisin. World Health Organization merekomendasikan Pyrosequencing, metode sequencing nukleotida yang dapat mendeteksi mutasi gen rpoB, gen katG dan/atau inhA penyandi resistansi Isoniazid. Tujuan penelitian ini adalah menentukan apakah kedua alat ini memberikan hasil interpretasi yang sama, apakah ada perbedaan deteksi posisi mutasi kodon gen rpoB dan apakah ditemukan mutasi gen katG dan/atau inhA. Penelitian ini merupakan studi observasional komparatif dengan rancangan cross-sectional. Subjek penelitian adalah pasien usia ≥18 tahun yang diinterpretasi RR-TB berdasarkan Xpert MTB/RIF assay di RSUP Dr. Hasan Sadikin Bandung dan belum mendapat pengobatan. Xpert MTB/RIF assay menginterpretasi 40 subjek Rifampicin-resistant TB sedangkan Pyrosequencing menginterpretasi 30 subjek MDR-TB, 9 subjek RR-TB dan satu subjek Isoniazid-resistant TB pada bulan Januari-Februari 2020. Deteksi posisi mutasi kodon gen rpoB antara Xpert MTB/RIF assay dan Pyrosequencing tidak berbeda bermakna (p=0,389). Pyrosequencing mendeteksi 27 mutasi gen katG, 3 mutasi gen inhA, satu mutasi gen katG dan inhA. Kesimpulan, Pyrosequencing dapat digunakan untuk deteksi resistansi Rifampisin dan Isoniazid pada MDR-TB secara lebih akurat.
Positivity Rate of Pyrosequencing to Diagnose Drug-Resistant Tuberculosis Directly from Sputum with Different Bacterial Load Selma Zein Syafira; Nabilla Ghina Zavitri; Su Yan; Yunia Sribudiani; Alexander Lezhava; Lidya Chaidir
The Indonesian Biomedical Journal Vol 12, No 4 (2020)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v12i4.1130

Abstract

BACKGROUND: Molecular techniques, which detect mutations associated with drug resistance tuberculosis (TB), are promising technologies for rapid diagnosis and monitoring of drug-resistant TB. Pyrosequencing is a potential rapid and robust molecular technique to detect drug resistance but its performance in clinical samples is less investigated. This study aimed to determine the positivity rate of pyrosequencing to diagnose drug-resistant TB directly from sputum samples with different grades of sputum smear microscopy results.METHODS: Thirty-five sputum specimens from drug-resistant TB suspects were submitted for acid-fast bacilli (AFB) microscopy. All specimens were cultured using microscopic observation drug susceptibility (MODS) culture. Pyrosequencing was performed to DNA extracted from sputum of culture-positive patients.RESULTS: MODS culture was positive in 19/35 subjects (54.29%) samples; 16 smear-positive and three smear-negative. Using pyrosequencing, Mycobacterium tuberculosis was identified in all culture-positive samples, including smear-negative samples. A complete resistance profile for 16 (82.35%) samples could be generated. Pyrosequencing failed to show results for eis or gyrA promoter in three samples. Nine of 19 patients were multidrug resistant-TB (MDR-TB), 1/19 was rifampicin-resistance TB (RR-TB), and 4/19 were pre-extensively drug-resistant TB (pre-XDR-TB). Two novel mutations in rpoB and rrs (associated with rifampicin and aminoglycoside, respectively) were found in this study.CONCLUSION: The results of this study demonstrates high positivity rates of pyrosequencing to detect drug-resistant TB directly from sputum samples with different grades of smear microscopy, as the surrogate of bacterial load. The assay can be used as a first prediction test of drug resistance prior to confirmation by phenotypic tests.KEYWORDS: drug-resistant tuberculosis, pyrosequencing, direct sputum
Perbandingan Masalah Mental Emosional dan Karakteristik Pasien Remaja dengan Talasemia Mayor di Poli Talasemia RSUP Dr. Hasan Sadikin Bandung Qory Aulia Sukma; Rodman Tarigan; Lynna Lidyana
Sari Pediatri Vol 23, No 1 (2021)
Publisher : Badan Penerbit Ikatan Dokter Anak Indonesia (BP-IDAI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14238/sp23.1.2021.23-7

Abstract

Latar belakang. Talasemia adalah penyakit genetik hematologik yang berdampak terhadap perubahan fisik dan kegiatan sehari-hari pasien sehingga berpengaruh pada psikososial pasien, terutama pasien remaja yang mengalami perubahan emosional pada fase transisi menuju dewasa.Tujuan. Penelitian dilakukan untuk mengetahui perbandingan masalah mental emosional dengan karakteristik pasien remaja talasemia mayor di Poli Talasemia RSUP Dr. Hasan Sadikin Bandung.Metode. Penelitian ini adalah studi deskriptif-analitik dengan desain potong lintang yang dilakukan pada bulan Agustus– September 2019. Subjek penelitian adalah 96 pasien talasemia di Poli Talasemia RSUP Dr. Hasan Sadikin Bandung dengan kelompok usia 11-17 tahun yang mengisi self-assessment Strengths and Difficulties Questionnaire (SDQ).Hasil. Dari 96 subjek penelitian, 49 (51%) adalah perempuan. Enam puluh empat (67%) adalah kelompok usia 11-14 tahun, dan 46 (48%) di antaranya adalah siswa SMP. Gambaran masalah mental emosional pasien meliputi masalah emosional 17 (18%), conduct 12 (12%), hiperaktivitas 8 (8%), dan masalah hubungan dengan teman sebaya 7 (7%). Kesimpulan. Tidak ada perbedaan antara masalah mental emosional dan karakteristik pasien remaja dengan talasemia mayor.
Application of Replicate Organism Detection and Counting Method (RODAC) in Measuring Mycobacterium Tuberculosis Contamination in High Burden Laboratories Lidya Chaidir; Neng Rina Susilawati; Mandala Ajie; Jessi Annisa; Muti'ah Nurul Jihadah
Althea Medical Journal Vol 9, No 4 (2022)
Publisher : Faculty of Medicine Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15850/amj.v9n4.2789

Abstract

Background: Technicians working in high burden tuberculosis (TB) laboratories pose a higher risk of being infected by Mycobacterium tuberculosis from clinical samples. Contamination control is mandatory to detect the release of bacteria into the working environment and to minimize the risk of exposure to the workers. The contamination measurement is rarely performed due to the lack of standard methodology. This study optimized and applied a unique culture-based method named Replicate Organism Detection and Counting (RODAC) plates to assess the presence of M. tuberculosis contaminant in the laboratory with high burden of clinical samples. Methods: RODAC was applied on twenty working surfaces in the Mycobacteriology Laboratory of Universitas Padjadjaran. The results of RODAC were compared with DNA-based detection from the same working surfaces using in-house IS6110 real-time PCR (IS6110-qPCR). The detection limit of the RODAC plate was 19.6 CFU mL-1.Results: From all working surfaces tested, two distinct colonies were found on RODAC plate stamped on the Ziehl-Neelsen staining basin. Those colonies were identified as M. tuberculosis and non-tuberculous mycobacteria (NTM), as confirmed by the MPT64 antigen test and the presence of acid-fast bacilli. IS6110-qPCR detected the presence of M. tuberculosis DNA in ten sampling points, including the ZN staining basin, incubators, and microscopy areas. IS6110-qPCR detected more working surface contamination versus RODAC. However, it was noted that RODAC, which was a culture-based method, detected live bacteria, while PCR could not distinguish between live and dead bacteria.Conclusion: The application of the RODAC plate is more suitable for monitoring the contamination of live bacteria in the working environment and to inform a proper corrective action.
Effective Visual Media to Increase Knowledge and Comprehension of Multidrug Resistant Tuberculosis Among Patients and Their Caregivers Lidya Chaidir; Dyshelly Nurkartika Pascapurnama; Claudia Selviyanti; Cindy Natasha; Muti'ah Nurul Jihadah; Prayudi Santoso
Majalah Kedokteran Bandung Vol 55, No 2 (2023)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15395/mkb.v55n2.2986

Abstract

Indonesia ranks thirdamong countries with the largest number of TB cases after India and China. Globally, more than 3-4% of all TB cases are Multidrug-Resistant Tuberculosis (MDR TB). MDR TB is a more complicated TB that needs extra treatments, which extend treatment time and increase adverse effects. Thus, MDR TB patients and their families often feel demotivated about completing treatment, leading to loss to follow up, which contributes to the never-ending transmission and greatly affects the success rate of the national TB control program. Thus, better knowledge and perception on MDR treatment for patients and families plays a crucial role in dealing with this issue. A cross-sectional study was performed from April to June 2019 to evaluate the effectiveness of visual educational media for TB patients and their caregivers. Participants consisted of 144 patients diagnosed with MDR TB and their caregivers or family members (n=22). A pre-test was administered before an education session by a nurse and visual media were used as the educational material. At the end of the session, a post-test was performed. The post-test score was then compared to the pre-test score to evaluate the session’s effectiveness using the paired t-test. Result showed significant increase in the post-test score (t= 3.249, df=3, p=0.04), with the caregivers attained a higher score, showing better improvement in knowledge after the session compared to the patient group. Hence, the MDR TB educational intervention using visual media is considered effective to increase participants ‘understanding of MDR TB. It is expected that with increased knowledge on MDR TB, the treatment success rate will increase and becomes the catalyst for the nationwide TB control strategy.
Clinical Performance of a Molecular-based Lateral Flow Assay for Rapid and Simultaneous Detection of Tuberculosis and Non-Tuberculous Mycobacteria in Indonesia Lidya Chaidir; Nabilla Ghina Zavitri; Adi Imam Cahyadi; Claudia Selviyanti; Dyshelly Nurkartika Pascapurnama; Jessi Annisa; Bachti Alisjahbana; Prayudi Santoso
Jurnal Respirologi Indonesia Vol 46 No 2 (2026)
Publisher : Perhimpunan Dokter Paru Indonesia (PDPI)/The Indonesian Society of Respirology (ISR)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.36497/jri.v46i2.1071

Abstract

Background: Diagnostic modalities that can differ Tuberculosis (TB) and Non-Tuberculosis Mycobacteria (NTM) are urgently needed, especially in TB high burden areas, to avoid misdiagnosis. A rapid molecular approach can be utilized to accommodate easy implementation for routine diagnosis in low-resource countries to decrease the turnaround time and increase sensitivity significantly. This study evaluated the diagnostic performance of PaxView® TB/NTM, a rapid multiplex PCR assay that utilizes universal lateral flow technology for the simultaneous detection and visualization of TB and NTM. Methods: The study was conducted in Bandung, Indonesia, using sputum samples from the outpatient TB clinic in Hasan Sadikin General Hospital. Sputum samples were collected consecutively from each patient. All the eligible samples underwent microbiological work-up and Xpert MTB/RIF testing with different sputum taken previously. The collected sputum was extracted, amplified, and then visualized using the PaxView® TB/NTM MPCR-ULFA kit. Results: The sensitivity and specificity of the PaxView® MPCR-ULFA were 83.0% and 92.1%, respectively. The positivity rate for MPCR ULFA showed significantly higher results (43.0%) compared to culture (32.8%), smear microscopy (26.3%), and was comparable with the Xpert MTB/RIF assay (43.5%). A total of 23 confirmed NTM-positive cases were found in this study (2.6%). Conclusion: The PaxView® MPCR-ULFA demonstrated promising performance for MTB and NTM detection. The sensitivity and specificity of this kit are comparable to those of other molecular diagnostic modalities. The simple platform using a test strip and straightforward visualization might be appropriately applied in poorly-equipped laboratories.
Co-Authors Adi Imam Cahyadi Alexander Lezhava Anna Tjandrawati Bachti Alisjahbana Cindy Natasha Claudia Selviyanti Claudia Selviyanti Coriejati Rita Dyshelly Nurkartika Pascapurnama Dyshelly Nurkartika Pascapurnama Ida Parwati Jessi Annisa Jessi Annisa Linda Choerunnisa Mandala Ajie Muti'ah Nurul Jihadah Muti'ah Nurul Jihadah Nabilla Ghina Zavitri Nabilla Ghina Zavitri Neng Rina Susilawati Prayudi Santoso Prayudi Santoso Qory Aulia Sukma Rodman Tarigan Selma Zein Syafira Su Yan Yunia Sribudiani