Rapid detection of brucella spp. using loop-mediated isothermal amplification (LAMP)

Shouyi Chen, Xunde Li, Juntao Li, Edward R. Atwill

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages99-108
Number of pages10
Volume1039
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1039
ISSN (Print)10643745

Keywords

  • Blood
  • Brucella
  • Detection
  • LAMP
  • Loop-mediated isothermal amplification
  • Milk

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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