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《Chinese Journal of Veterinary Medicine》 2012-02
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Establishment of a Cycling probe real-time PCR assay for Brucella detection

XU Zou-liang1,2,NAN Wen-long1,LU Ming-zhe1,ZHOU Jie1,TAN Peng-fei1,2, CHEN Yi-ping1,MAO Kai-rong3(1.Diagnostic Reagent Laboratory,China Animal Health and Epidemiology Center,Qingdao 266032 China; 2.College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China; 3.China Institute of Veterinary Drug Control,Beijing 100081,China)  
A Cycling real-time PCR assay was developed for the detection of all species and bivors of Brucella.According to the sequences of BCSP31 gene,a probe and two primers were designed.The sensitivity and standard curve of the Cycling real-time PCR assay were tested.All results showed that the detection limit of the assay was about 10 copies of the recombinant plasmid and the standard curve was favorable with 0.999 linear correlation coefficient.The specificity was perfect with typical "S" curve amplification of Brucella and negative results for other four irrelevant bacterias.Furthermore,the detection of 40 clinical samples showed 35 positive samples were detected by the Cycling probe real-time PCR,which included all 27 B4/B5-PCR positive samples.The coincidence rate of two methods was 80.0%.All results suggested that the Cycling probe real-time PCR assay was specific,sensitive and stable,which could be used to diagnose clinical samples.
【Fund】: “十一五”国家支撑计划(2006BAD06A12);; 公益性行业(农业)项目(200903027)
【CateGory Index】: S855.12
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Chinese Journal Full-text Database 6 Hits
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