title: Development of an in-house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients.
abstract: A number of nucleic acid amplification assays (NAAs) have been employed to detect tubercle bacilli in clinical specimens for tuberculosis (TB) diagnosis. Among these, loop-mediated isothermal amplification (LAMP) is an NAA possessing superior isothermal reaction characteristics. In the present study, a set of six specific primers targeting the Mycobacterium tuberculosis 16S rRNA gene with high sensitivity was selected and a LAMP system (MTB-LAMP) was developed. Using this system, a total of 200 sputum samples from Nepalese patients were investigated. The sensitivity of MTB-LAMP in culture-positive samples was 100 % (96/96), and the specificity in culture-negative samples was 94.2 % (98/104, 95 % confidence interval 90.5-97.9 %). The positive and negative predictive values of MTB-LAMP were 94.1 and 100 %, respectively. These results indicate that this MTB-LAMP method may prove to be a powerful tool for the early diagnosis of TB.
authors: Everest International Clinic and Research Center, Kathmandu, Nepal.|Sukraraj Tropical and Infectious Disease Hospital, Kathmandu, Nepal.
authors: Basu Dev
authors: BD
authors: Pandey
authors: Department of Microbiology, Tribhuban University, Kathmandu, Nepal.
authors: Ajay
authors: A
authors: Poudel
authors: Department of Infectious Diseases, Osaka Prefectural Institute of Public Health, Japan.
authors: Tomoko
authors: T
authors: Yoda
authors: Department of Infectious Diseases, Osaka Prefectural Institute of Public Health, Japan.
authors: Aki
authors: A
authors: Tamaru
authors: Asahi Kasei Co. Ltd, Shizuoka, Japan.
authors: Naozumi
authors: N
authors: Oda
authors: Research Center for Zoonosis Control, Hokkaido University, Hokkaido, Japan.
authors: Yukari
authors: Y
authors: Fukushima
authors: Department of Microbiology, Tribhuban University, Kathmandu, Nepal.
authors: Binod
authors: B
authors: Lekhak
authors: Department of Microbiology, Tribhuban University, Kathmandu, Nepal.
authors: Basista
authors: B
authors: Risal
authors: Everest International Clinic and Research Center, Kathmandu, Nepal.
authors: Bishnu
authors: B
authors: Acharya
authors: Department of Research and Biotechnology, Anandaban Hospital, Kathmandu, Nepal.
authors: Bishwa
authors: B
authors: Sapkota
authors: Research Center for Zoonosis Control, Hokkaido University, Hokkaido, Japan.
authors: Chie
authors: C
authors: Nakajima
authors: Section of Bacterial Infections, Research Collaboration Center on Emerging and Re-emerging Infections, Osaka University, Nonthaburi, Thailand.
authors: Tooru
authors: T
authors: Taniguchi
authors: National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand.
authors: Benjawan
authors: B
authors: Phetsuksiri
authors: Research Center for Zoonosis Control, Hokkaido University, Hokkaido, Japan.
authors: Yasuhiko
authors: Y
authors: Suzuki
mesh_terms: D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification*; D009390:Nepal; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011237:Predictive Value of Tests; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology*; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S