1种布鲁氏菌微滴式数字PCR检测方法的建立

doi:10.11843/j.issn.0366-6964.2021.06.030

Abstract

Abstract: This experiment was conducted to establish a droplet digital PCR (ddPCR) method for detection of Brucella. A ddPCR method was developed, the primers and probes were designed based on the conservative regions of BCSP31 gene of Brucella. The concentration of primer and probe, the annealing temperature were optimized. The sensitivity, specificity and reproducibility of ddPCR method were evaluated. In results, the optimal primer concentration and the probe concentration were 900 nmol·L^-1 and 250 nmol·L^-1, the optimum annealing temperature was 58℃. The detection limit of ddPCR method was 1.12 copies·μL^-1 with a good linear response, it had no cross reaction with E. coli O:157 and other four bacterium, both the intra-batch reproducibility and the inter-batch reproducibility were good. All the results showed that Brucella ddPCR was sensitive and specific, it was suitable for quantitative detection of clinical samples infected with Brucella.

Key words: Brucella, droplet digital PCR, detection method

CLC Number:

S852.614

MEI Li, WANG Yingchao, CHENG Rujia, YU Guoji, FAN Xuezheng, GAO Xiaolong, GAO Min, QIN Yuming, LI Xiaoying, LI Qiaoling, ZHU Liangquan, FENG Xiaoyu. A Droplet Digital PCR Method for Detection of Brucella[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1753-1759.

References 17

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A Droplet Digital PCR Method for Detection of Brucella

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