1种检测新城疫病毒的微滴式数字PCR方法

doi:10.11843/j.issn.0366-6964.2020.12.028

Abstract

Abstract: This experiment was conducted to establish a droplet digital PCR (ddPCR) method for the detection of Newcastle disease virus (NDV). A ddPCR method was developed, which the primers and probes were designed based on the conservative regions of F gene of NDV. The concentration of primer and probe, the annealing temperature in ddPCR reaction were optimized. The sensitivity, specificity and reproducibility of ddPCR method were evaluated. In results, the optimal primer concentration and the probe concentration were 900 and 250 nmol·L^-1, the optimum annealing temperature was 55℃. The detection limit of ddPCR method was 1.8 copies·μL^-1 with a good linear response, it had no cross reaction with other six viruses (include IBV), the coefficient of variation of sample repetition was 2.4%. All the results showed that NDV ddPCR was sensitive and specific, it was suitable for quantitative detection of clinical samples infected with NDV.

Key words: Newcastle disease virus, droplet digital PCR, detection method

CLC Number:

S855.3

MEI Li, WANG Yingchao, WU Di, LI Yue, SONG Yanjun, WEI Haitao, WANG Lin, GAO Xiaolong, FENG Xiaoyu. A Droplet Digital PCR Method for Detection of Newcastle Disease Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(12): 3187-3192.

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A Droplet Digital PCR Method for Detection of Newcastle Disease Virus

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