Loop-mediated isothermal amplification assay targeting the mpb70 gene for rapid differential detection of Mycobacterium bovis.
Loop-mediated isothermal amplification (LAMP) is a highly sensitive, rapid, cost-effective nucleic acid amplification method. Tuberculosis (TB) is widely popular in the world and it is difficult to cure. The fundamental treatment is to clear the types of TB pathogens such as Mycobacterium bovis (M. bovis), Mycobacterium tuberculosis (M. tuberculosis). In order to detect and diagnose TB early, we constructed the differential diagnostic method of TB. In this study, we used LAMP for detection of M. bovis, based on amplification of the mpb70 gene which is a unique gene in M. bovis strain. The LAMP assay was able to detect only seven copies of the gene per reaction, whereas for the conventional PCR, it was 70 copies. The LAMP was evaluated for its specificity using six strains of five Mycobacterium species and 18 related non-Mycobacterium microorganism strains as controls. The target three Mycobacterium strains were all amplified, and no cross-reaction was found with 18 non-Mycobacterium microorganism strains. TB was detected by two methods, LAMP and conventional PCR (based on mpb70 gene); the positive rates of the two methods were 9.55 and 7.01 %, respectively. Our results indicate that the LAMP method should be a potential tool with high convenience, rapidity, sensitivity and specificity for the diagnosis of TB caused by M. bovis. Most importance is that the use of LAMP as diagnostic method in association with diagnostic tests based on mpb70 gene would allow the differentiation between M. bovis and other Mycobacterium in humans or animals. The LAMP method is actually in order to detect human TB, and it can be used for differential diagnosis in this paper.
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.|Co-Innovation Center for Zoonotic Infectious Diseases in the western region, Shihezi, Xinjiang Province, 832000, People's Republic of China.
Hui
H
Zhang
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
Zhen
Z
Wang
Co-Innovation Center for Zoonotic Infectious Diseases in the western region, Shihezi, Xinjiang Province, 832000, People's Republic of China.|School of Medicine, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
Xudong
X
Cao
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
Zhengrong
Z
Wang
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.|Co-Innovation Center for Zoonotic Infectious Diseases in the western region, Shihezi, Xinjiang Province, 832000, People's Republic of China.
Jinliang
J
Sheng
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.|Co-Innovation Center for Zoonotic Infectious Diseases in the western region, Shihezi, Xinjiang Province, 832000, People's Republic of China.
Yong
Y
Wang
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
Jing
J
Zhang
School of Life Sciences, Shangqiu Normal University, Shangqiu, Henan province, 476000, People's Republic of China.
Zhiqiang
Z
Li
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China. guxinlidky@sohu.com.
Xinli
X
Gu
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China. ccf_xb@126.com.|Co-Innovation Center for Zoonotic Infectious Diseases in the western region, Shihezi, Xinjiang Province, 832000, People's Republic of China. ccf_xb@126.com.
Chuangfu
C
Chen
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001431:Bacteriological Techniques; D006801:Humans; D009163:Mycobacterium bovis / Q000235:genetics* / Q000302:isolation & purification*; D009169:Mycobacterium tuberculosis / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014376:Tuberculosis / Q000175:diagnosis* / Q000382:microbiology
D001426:Bacterial Proteins; C060216:MPB70 protein, Mycobacterium bovis
LAMP; M. bovis; M. tuberculosis; mpb70