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KMID : 1023520110340040333
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Korean Journal of Veterinary Service
2011 Volume.34 No. 4 p.333 ~ p.339
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Development of Loop-mediated isothermal amplification method for the rapid and sensitive detection of bovine tuberculosis in Korea native cattle
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Hwang Eun-Suk
Lee Tae-Uk
Jung Dae-Young
Cho Ho-Seong
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Abstract
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Loop-mediated isothermal amplification (LAMP) was developed to detect Mycobacterium tuberculosis complex (MTC) and non-tuberculous mycobacterium (NTM) genomic DNA in blood samples of Korea native cattle. A set of four primers, two outer and two inner, were designed from M. bovis and M. avium genomic DNA targeting the IS6110 and 16S rRNA gene, respectively. Based on 85 Intradermal Tuberculin Test (ITT) positive blood sample and using conventional PCR and LAMP, the agreement quotient (kappa), which measures agreement beyond chance were 0.93 (conventional PCR) and 0.97 (LAMP), respectively. The detection limit of the LAMP method was 2.0×102 copy/ml M. bovis and M. avium cells, compared to 2.0×103 copy/ml M. bovis and M. avium cells for conventional PCR. These results suggest that the LAMP is a powerful tool for rapid, sensitive, and practical detection of MTC and NTM in blood samples of Korea native cattle.
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KEYWORD
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Bovine tuberculosis, Loop-mediated isothermal amplification (LAMP), Mycobacterium bovis
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