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Rapid Detection of Brucella spp. Using Loop-Mediated Isothermal Amplification (LAMP)

  • Shouyi Chen
  • Xunde Li
  • Juntao Li
  • Edward R. Atwill
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1039)

Abstract

Brucella spp. are facultative intracellular bacteria that cause zoonotic disease of brucellosis worldwide. Livestock that are most vulnerable to brucellosis include cattle, goats, and pigs. Brucella spp. cause serious health problems to humans and animals and economic losses to the livestock industry. Traditional methods for detection of Brucella spp. take 48–72 h (Kumar et al., J Commun Dis 29:131–137, 1997; Barrouin-Melo et al., Res Vet Sci 83:340–346, 2007) that do not meet the food industry’s need of rapid detection. Therefore, there is an urgent need of fast, specific, sensitive, and inexpensive method for diagnosing of Brucella spp. Loop-mediated isothermal amplification (LAMP) is a method to amplify nucleic acid at constant temperatures. Amplification can be detected by visual detection, fluorescent stain, turbidity, and electrophoresis. We targeted at the Brucella-specific gene omp25 and designed LAMP primers for detection of Brucella spp. Amplification of DNA with Bst DNA polymerase can be completed at 65 °C in 60 min. Amplified products can be detected by SYBR Green I stain and 2.0 % agarose gel electrophoresis. The LAMP method is feasible for detection of Brucella spp. from blood and milk samples.

Key words

Loop-mediated isothermal amplification LAMP Detection Brucella Milk Blood 

Notes

Acknowledgment

This work was partially supported by Guangzhou Medicine and Hygiene Foundation No. 2006-zdi-11.

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Shouyi Chen
    • 1
  • Xunde Li
    • 2
  • Juntao Li
    • 1
  • Edward R. Atwill
    • 2
  1. 1.Department of MicrobiologyGuangzhou Center for Disease Control and PreventionGuangzhouChina
  2. 2.Department of Population Health and ReproductionUniversity of California DavisDavisUSA

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