Riemerella anatipestifer detection kit and method based on loop-mediated isothermal amplification technology
A ring-mediated isothermal and Riemerella anatipestifer technology is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microbial measurement/inspection. Achieve the effect of simple and fast operation, strong specificity and low detection cost
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[0035] Example 1
[0036] 1. A detection kit for Riemerella anatipestifer based on loop-mediated isothermal amplification technology, consisting of the following reagents:
[0037] a. The upstream inner primer FIP aqueous solution and the downstream inner primer BIP aqueous solution each with a concentration of 5 μmol / L, the upstream outer primer F3 aqueous solution and the downstream outer primer B3 aqueous solution with a concentration of 20 μmol / L each: the primer sequence is as follows:
[0038] Upstream inner primer FIP: 5’-caggcttccacccattgtccaa-tgagacacggaccagactc-3’;
[0039] Downstream inner primer BIP: 5’-cgtgaaggacgacggccctat-ctaccctcacgagagtaggt-3’;
[0040] Upstream outer primer F3: 5’-tttagggggcctgagagg-3’;
[0041] Downstream outer primer B3: 5'-ccggtgcttattcgtacagt-3';
[0042] b. A Bst DNA polymerase aqueous solution with a concentration of 8U / μl;
[0043] c. 10× thermal polymerization buffer solution: Tris-HCl with a concentration of 250mmol / L, pH 8.8, potassium chloride ...
Example Embodiment
[0053] Example 2
[0054] 1. The Riemerella anatipestifer detection kit based on the loop-mediated isothermal amplification technology is different from the kit described in Example 1 in the specific primer sequence. The primer sequence of this kit is as follows:
[0055] Upstream inner primer FIP: 5’-tattcctcactgctgcctcccg-gtgatcccccacactggt-3’;
[0056] Downstream inner primer BIP: 5’-cgtgaaggacgacggccctat-ctaccctcacgagagtaggt-3’;
[0057] Upstream outer primer F3: 5’-tttagggggcctgagagg-3’;
[0058] Downstream outer primer B3: 5'-ccggtgcttattcgtacagt-3'.
[0059] 2. The method for detecting Riemerella anatipestifer using the ring-mediated isothermal amplification technology-based Riemerella anatipestifer detection kit is the same as the method described in Example 1. As a result, it is found that the color of the blank control group It is yellow, indicating that it does not contain Riemerella anatipestifer; the color of the experimental group changes to green, indicating that it conta...
Example Embodiment
[0060] Example 3
[0061] 1. The Riemerella anatipestifer detection kit based on the loop-mediated isothermal amplification technology is different from the kit described in Example 1 in the specific primer sequence. The primer sequence of this kit is as follows:
[0062] Upstream inner primer FIP: 5’-caggcttccacccattgtccaa-acactggtactgagacacgg-3’;
[0063] Downstream inner primer BIP: 5’-cgtgaaggacgacggccctat-ctaccctcacgagagtaggt-3’;
[0064] Upstream outer primer F3: 5’-tttagggggcctgagagg-3’;
[0065] Downstream outer primer B3: 5'-ccggtgcttattcgtacagt-3'.
[0066] 2. The method for detecting Riemerella anatipestifer using the ring-mediated isothermal amplification technology-based Riemerella anatipestifer detection kit is the same as the method described in Example 1. As a result, it is found that the color of the blank control group It is yellow, indicating that it does not contain Riemerella anatipestifer; the color of the experimental group changes to green, indicating that it con...
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