Method for detecting mycoplasma synoviae and reagent used in method

[0042] Example 1. Materials and Methods

[0043] 1.1. Virus strains

[0044] The strains of Mycoplasma gallis synoviae (MS K1415) and Mycoplasma gallis synovium strains (MS 28) are kept by our laboratory, and are in the literature "Deng Xianwen et al., Establishment of a rapid visual detection method for Mycoplasma gallis synovium LAMP, Chinese Journal of Preventive Veterinary Medicine, 2014, 36(1): 46-49.”, the public can obtain the biological material from the applicant in accordance with the relevant provisions of the national biosafety, and the biological material is only used for repeating the relevant experiments of the present invention and cannot be used for other purposes. .

[0045] Infectious bronchitis virus (IBV Mass 41), Mycoplasma gallisepticum (MG S6), Newcastle disease virus (NDVLaSota), avian reovirus (Reo S1133), avian influenza H9N2 066C strain (AIV H9N2 066C), by our laboratory Preserved, avian metapneumonia virus (APV / MN-10) was donated by Professor Lu fr...

[0074] Example 2. Results

[0075] 2.1. Optimization of annealing temperature for ddPCR reaction

[0076] In the ddPCR reaction, the upstream and downstream primer concentrations and probe concentrations were 20 μmol / μL and 10 μmol / μL, respectively, and the template concentration was about 350 copies / μL, and the annealing temperature was set to 50, 51, 52, 54, 55, 56, 58 A total of 8 different gradients at 60°C were used for ddPCR amplification. When the annealing temperature was 54°C, the number of positive droplets was the highest, which was 326 copies / μL. The result is as figure 1 shown, figure 1 Among them, A11 represents 50°C, B11 represents 51°C, C11 represents 52°C, D11 represents 54°C, E11 represents 55°C, F11 represents 56°C, G11 represents 58°C, and H11 represents 60°C. figure 1 The abscissa of the middle and left figure represents the number of droplets, and the ordinate represents the fluorescence intensity; figure 1 The abscissa of the middle and right figures...