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Development of a Multiplex PCR for Detection of Brucella abortus， Brucella melitensis，Brucella suis and Brucella canis

CHEN Si1，WANG Xiu-ran1，QIAN Jing1，WANG Xiao-xu1，LANG Xu-long1，2，WANG Xing-long1，2   

  1. （1.Institute of Military Veterinary，Academy of Military Medical Sciences，Changchun 130122，China； 2.Key Laboratory of Jilin Province for Zoonosis Prevention and Control，Changchun 130122，China）
  • Received:2013-09-05 Online:2014-02-20 Published:2014-03-27

Abstract: The purpose of the experiment was to establish a rapid multiplex PCR detection method which could distinguish B.abortus，B.melitensis，B.suis and B.canis. According to the differences of IS711 and complete genome sequences，four pairs of primers were designed. Multiplex PCR reaction system and conditions were optimized，the specificity，sensitivity and stability of the multiplex PCR were analyzed.Through the establishment of the multiplex PCR，B.abortus，B. melitensis，B. suis and B.canis could amplify the expected fragment，the sizes of the expected fragment were 494，732，591 and 272 bp，respectively. The PCR sensitivity of B.abortus，B.melitensis，B.suis and B.canis were 1.1×102，5.1×102，3.5×102 and 2.5×102 CFU/mL，respectively. Detected artificially infectious samples of milk by PCR，PCR sensitivity could reach 1.0×103 CFU/mL.The developed multiplex PCR method was simple，fast，high sensitivity，and had good prospects and important significance for the identification of B.abortus，B.melitensis，B.suis and B.canis.

Key words: Brucella; multiplex PCR; detection