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：99

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

来源：

JOURNAL OF MEDICAL MICROBIOLOGY | 2008年 / 57卷 / 04期

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

页码：439 / 443

页数：5

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