Sensitive and rapid detection of Mycoplasma capricolum subsp. capripneumoniae by Loop-mediated isothermal amplification

HE Ying; ZHANG Nian-zhang; ZHAO Ping; CHU Yue-feng; GAO Peng-cheng; ZHANG Jian-jun,; LIU Xiang-tao and; Lu Zhong-xin*

doi:10.5897/AJB12.2960

African Journal of
Biotechnology

Abbreviation: Afr. J. Biotechnol.
Language: English
ISSN: 1684-5315
DOI: 10.5897/AJB
Start Year: 2002
Published Articles: 12487

Full Length Research Paper

Sensitive and rapid detection of Mycoplasma capricolum subsp. capripneumoniae by Loop-mediated isothermal amplification

HE Ying

HE Ying
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Epizootic Diseases of Grazing Animal of Ministry of Agriculture, Lanzhou 730046, China.
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ZHANG Nian-zhang

ZHANG Nian-zhang
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Epizootic Diseases of Grazing Animal of Ministry of Agriculture, Lanzhou 730046, China.
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ZHAO Ping

ZHAO Ping
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Epizootic Diseases of Grazing Animal of Ministry of Agriculture, Lanzhou 730046, China.
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CHU Yue-feng

CHU Yue-feng
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Epizootic Diseases of Grazing Animal of Ministry of Agriculture, Lanzhou 730046, China.
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GAO Peng-cheng

GAO Peng-cheng
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Epizootic Diseases of Grazing Animal of Ministry of Agriculture, Lanzhou 730046, China.
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ZHANG Jian-jun,

ZHANG Jian-jun,
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Epizootic Diseases of Grazing Animal of Ministry of Agriculture, Lanzhou 730046, China.
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LIU Xiang-tao and

LIU Xiang-tao and
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Epizootic Diseases of Grazing Animal of Ministry of Agriculture, Lanzhou 730046, China.
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Lu Zhong-xin*

Lu Zhong-xin*
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Epizootic Diseases of Grazing Animal of Ministry of Agriculture, Lanzhou 730046, China.
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Article Number - DE4E07F44747
Vol.13(21), pp. 2113-2118 , May 2014
https://doi.org/10.5897/AJB12.2960

Received: 20 November 2012
Accepted: 02 August 2013
Published: 21 May 2014

Abstract

A set of four specific primers was designed by targeting the H2 gene sequences of Mycoplasma capricolum subsp. capripneumoniae (MCCP). Using Bst DNA polymerase, the products were amplified for 60 min at 65°C in a simple water bath. Compared with a polymerase chain reaction (PCR) test that targets the H2 gene sequences of MCCP, the sensitivity of the loop-mediated isothermal amplification (LAMP) assay was higher (approximately 0.75 fg DNA per reaction). The LAMP products could be visualized by agar gel electrophoresis. There were no cross reactions with other strains in the Mycoplasma mycoides cluster, which indicates the high specificity of the LAMP procedure. The LAMP assay was able to detect MCCP in tissue.

Key words: Mycoplasma capricolum subsp. Capripneumoniae, loop-mediated isothermal amplification, rapid detection.

Abbreviation

Abbreviations: CCPP, Contagious caprine pleuropneumonia; MCCP, Mycoplasma capricolum subsp. capripneumoniae; LAMP, loop-mediated isothermal amplification; PCR, polymerase chain reaction.