Establishment and evaluation of rapid discriminate PCR assay for Brucella abortus, Brucella melitensis and Brucella suis
LIU Zhi-guo1, 2, TA Na2, LI Xiao-yan2, WANG Miao3, ZHAO Hong-yan1, PIAO Dong-ri1, YU Rui-ping2, MI Jing-chuan2, LI Zhen-jun1
1. National Institute of Infectious Diseases Control and Prevention, Chinese Center for Disease Control and Prevention/Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease, Beijing 102206, China; 2. Inner Mongolia Autonomous Region Central for Comprehensive Disease Control and Prevention, Huhhot 010031, China; 3.Ulanqab Center for Endemic Disease Control and Prevention, Ulanqab 012000, China
Abstract：The aim of this study is to establish a rapid PCR discriminate assay for simultaneous identified Brucella genus, B. abortus, B. melitensis and B. suis in single reaction system. Amplification primers of Brucella genus and B. abortus, B. melitensis, and B. suis were combined and optimizated, a kind of novelty BAMS-PCR was established. Subsequently, specificity and sensitivity of BAMS-PCR were evaluated. In present study, 219 Brucella spp. were identified for assessed practicable of BAMS-PCR. Finally, DNA of Brucella clinical sample were detected by BAMS-PCR for evaluated using valuable in clinical diagnosis of Brucellosis. BAMS-PCR can simultaneous identified Brucella genus, B. abortus (bv.1,2 and 4), B. melitensis (bv.1-3) and B. suis (bv.1) in single reaction system, and this method had very higher specificity and sensitivity, detecting sensitivity of primers of Brucella genus, B. abortus, B. melitensis and B. suis with 10 pg/μL,100 pg/μL, 10 pg/μL and 100 pg/μL, respectively. There were 100% identification coincidence rate between BAMS-PCR and convention biotypes testing based on amplificated results of 219 Brucella spp. BAMS-PCR detected showing that there were only 6 serum samples were positive, others (whole blood and tissues (sheep spleen)) samples were all negative. BAMS-PCR was a simple, rapid, high efficiency and accuracy identification assay for Bruclla spp., could as first choice discriminate method of clinical isolates Bruclla spp.
刘志国, 塔娜, 李晓燕, 王妙, 赵鸿雁, 朴东日, 蔚瑞平, 米景川, 李振军. 牛羊猪种布鲁氏菌快速鉴别PCR方法的建立及评价[J]. 中国人兽共患病学报, 2019, 35(5): 399-403.
LIU Zhi-guo, TA Na, LI Xiao-yan, WANG Miao, ZHAO Hong-yan, PIAO Dong-ri, YU Rui-ping, MI Jing-chuan, LI Zhen-jun. Establishment and evaluation of rapid discriminate PCR assay for Brucella abortus, Brucella melitensis and Brucella suis. Chinese Journal of Zoonoses, 2019, 35(5): 399-403.
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2019, Vol. 35 
