Development of an in-house loop-mediated isothermal amplification (LAMP) assay for detection of Mycobacterium tuberculosis and evaluation in sputum samples of Nepalese patients

被引:96
作者
Pandey, Basu Dev [2 ,3 ]
Poudel, Ajay [4 ]
Yoda, Tomoko [5 ]
Tamaru, Aki [5 ]
Oda, Naozumi [6 ]
Fukushima, Yukari [1 ]
Lekhak, Binod [4 ]
Risal, Basista [4 ]
Acharya, Bishnu [3 ]
Sapkota, Bishwa [7 ]
Nakajima, Chie [1 ]
Taniguchi, Tooru [8 ]
Phetsuksiri, Benjawan [9 ]
Suzuki, Yasuhiko [1 ]
机构
[1] Hokkaido Univ, Res Ctr Zoonosis Control, Sapporo, Hokkaido 060, Japan
[2] Sukraraj Trop & Infect Dis Hosp, Kathmandu, Nepal
[3] Everest Int Clin & Res Ctr, Kathmandu, Nepal
[4] Tribhuban Univ, Dept Microbiol, Kathmandu, Nepal
[5] Osaka Prefectural Inst Publ Hlth, Dept Infect Dis, Osaka, Japan
[6] Asahi Kasei Co Ltd, Shizuoka, Japan
[7] Anandaban Hosp, Dept Res & Biotechnol, Kathmandu, Nepal
[8] Osaka Univ, Res Collaborat Ctr Emerging & Reemerging Infect, Sect Bacterial Infect, Nonthaburi, Thailand
[9] Natl Inst Hlth, Dept Med Sci, Minist Publ Hlth, Nonthaburi, Thailand
关键词
D O I
10.1099/jmm.0.47499-0
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页码:439 / 443
页数:5
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