Development of an in-house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients.
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.
Pandey
Basu Dev
BD
Everest International Clinic and Research Center, Kathmandu, Nepal.|Sukraraj Tropical and Infectious Disease Hospital, Kathmandu, Nepal.
Poudel
Ajay
A
Department of Microbiology, Tribhuban University, Kathmandu, Nepal.
Yoda
Tomoko
T
Department of Infectious Diseases, Osaka Prefectural Institute of Public Health, Japan.
Tamaru
Aki
A
Department of Infectious Diseases, Osaka Prefectural Institute of Public Health, Japan.
Oda
Naozumi
N
Asahi Kasei Co. Ltd, Shizuoka, Japan.
Fukushima
Yukari
Y
Research Center for Zoonosis Control, Hokkaido University, Hokkaido, Japan.
Lekhak
Binod
B
Department of Microbiology, Tribhuban University, Kathmandu, Nepal.
Risal
Basista
B
Department of Microbiology, Tribhuban University, Kathmandu, Nepal.
Acharya
Bishnu
B
Everest International Clinic and Research Center, Kathmandu, Nepal.
Sapkota
Bishwa
B
Department of Research and Biotechnology, Anandaban Hospital, Kathmandu, Nepal.
Nakajima
Chie
C
Research Center for Zoonosis Control, Hokkaido University, Hokkaido, Japan.
Taniguchi
Tooru
T
Section of Bacterial Infections, Research Collaboration Center on Emerging and Re-emerging Infections, Osaka University, Nonthaburi, Thailand.
Phetsuksiri
Benjawan
B
National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand.
Suzuki
Yasuhiko
Y
Research Center for Zoonosis Control, Hokkaido University, Hokkaido, Japan.
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
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S