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Detection of Mycobacterium bovis in environmental samples using nested PCR KCI 등재

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  • URLhttps://db.koreascholar.com/Article/Detail/406270
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예방수의학회지 (Journal of Preventive Veterinary Medicine)
한국예방수의학회(구 한국수의공중보건학회) (The Korean Society of Preventive Veterinary Medicine)
초록

The detection of Mycobacterium bovis (M. bovis) in environmental samples with precision is imperative to control bovine tuberculosis (bTB) infections at the herd level, as residual M. bovis remains one of the major causes of recurring infections. In this study, a nested PCR method for the detection of M. bovis in environmental samples was applied to identify potential environmental reservoirs of the bacterium. A set of 200 environmental samples (167 fecal samples and 33 water samples) from 39 herds with a history of bTB outbreak was analyzed using a nested PCR method to detect residual M. bovis. Amplicon libraries of the IS6110 target gene fragment were amplified from M. bovis DNA using two established primer sets. A positive nested PCR result was observed in 69.5% of fecal samples and 66.7% of water samples, thus showing that residual M. bovis was present in the environmental samples of bTB-positive herds in a high proportion. This study is the first to demonstrate high levels of M. bovis DNA in environmental samples and to show that environmental reservoirs of this pathogen contribute to recurring outbreaks of bTB. Environmental monitoring of herds in which bTB outbreaks have occurred with high sensitivity and specificity is expected to help prevent the recurrence of potential bTB disease and improve the herd environment.

목차
INTRODUCTION
MATERIALS AND METHODS
    Investigation of the characteristics of past bTBoutbreaks in Korea from 2012 to 2017
    Sample collection
    Culture of M. bovis from environmental samples
    Genomic DNA sample preparation
    Nested PCR conditions
    Sequencing of PCR products
    Confirmation of sensitivity and specificity of nested PCR
    Typing of M. bovis cultured and isolated from environmentalsamples
RESULTS
    Characteristics of previous bTB outbreaks in Koreafrom 2012-2017
    Environmental sample cultures
    Sensitivity and specificity of nested PCR in environmentalsamples
    Detection of M. bovis in environmental samples usingnested PCR
    Application of nested PCR to environmental samplesobtained from different locations
DISCUSSION
ACKNOWLEDGEMENTS
DISCLOSURE
REFERENCES
APPENDICES
저자
  • Tae-Woon Kim(Bacterial Disease Division, Animal and Plant Quarantine Agency)
  • Yun-Ho Jang(Bacterial Disease Division, Animal and Plant Quarantine Agency)
  • Min Kyu Jeong(Bacterial Disease Division, Animal and Plant Quarantine Agency)
  • Jae Myung Kim(Bacterial Disease Division, Animal and Plant Quarantine Agency)
  • Young Ju Lee(College of Veterinary Medicine, Kyungpook National University)
  • Seung Bum Kim(Department of Microbiology and Molecular Biology, Chungnam National University)
  • Soon-Seek Yoon(Bacterial Disease Division, Animal and Plant Quarantine Agency)
  • Jeong-Soo Choi(Bacterial Disease Division, Animal and Plant Quarantine Agency) Corresponding Author