Salmonella enteritidis detection reagent kit and method based on loop-mediated isothermal amplification technology
A Salmonella Enteritidis, ring-mediated isothermal technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems that have not been seen in the detection of Salmonella Enteritidis, and achieve simple and fast operation and low detection cost The effect of low and high sensitivity
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Embodiment 1
[0037] 1. A detection kit for Salmonella enteritidis based on loop-mediated isothermal amplification technology, consisting of the following reagents:
[0038] a. The upstream internal primer FIP aqueous solution and the downstream internal primer BIP aqueous solution with a concentration of 5 μmol / L, the upstream external primer F3 aqueous solution and the downstream external primer B3 aqueous solution with a concentration of 20 μmol / L: the primer sequences are as follows:
[0039] Upstream inner primer FIP: 5'-ttccagaacatgctcgctgc-ttgatgtggttggttcgtc-3'
[0040] Downstream internal primer BIP: 5'-tatagctcattctgacctctaagcc-gatatactccctgaatctgagaa-3';
[0041] Upstream outer primer F3: 5'-tagccaaaagaaaaccgcc-3';
[0042] Downstream outer primer B3: 5'-caggctgattactaaacctt-3';
[0043] b. An aqueous solution of Bst DNA polymerase with a concentration of 8 U / μl;
[0044] c, 10× thermal polymerization reaction buffer: Tris-HCl with a concentration of 250mmol / L and a pH of 8.8,...
Embodiment 2
[0055] 1. The Salmonella Enteritidis detection kit based on the loop-mediated isothermal amplification technology is different from the kit described in Example 1 in that the specific primer sequences are different. The primer sequences of this kit are as follows:
[0056] Upstream inner primer FIP: 5'-ctgagaaagaaaaactcattgaccg-ctggaaagcctctttatatagct-3';
[0057] Downstream internal primer BIP: 5'-aaggtttagtaaatcagcctgttgt-atgacaacttcgaaggtgg-3';
[0058] Upstream outer primer F3: 5'-tagccaaaagaaaaccgcc-3';
[0059] Downstream outer primer B3: 5'-tcgttcttctggtacttacg-3'.
[0060] 2. The method for detecting Salmonella enteritidis using the Salmonella enteritidis detection kit based on the loop-mediated isothermal amplification technology is the same as the method described in Example 1. It was found that the colors of the control groups 1 to 8 and the blank control group were both Yellow means that it does not contain Salmonella Enteritidis; the color of the experimental gr...
Embodiment 3
[0062] 1. The Salmonella Enteritidis detection kit based on the loop-mediated isothermal amplification technology is different from the kit described in Example 1 in that the specific primer sequences are different. The primer sequences of this kit are as follows:
[0063] Upstream inner primer FIP: 5'-catgctcgctgcacaaaagc-gagaggcggtttgatgtg-3';
[0064] Downstream inner primer BIP: 5'-ctggaaagcctctttatatagctca-tgatatactccctgaatctgaga-3';
[0065] Upstream outer primer F3: 5'-gggaggagctttagccaa-3';
[0066] Downstream outer primer B3: 5'-atggtgagcagacaacag-3'.
[0067] 2. The method for detecting Salmonella enteritidis using the Salmonella enteritidis detection kit based on the loop-mediated isothermal amplification technology is the same as the method described in Example 1. It was found that the colors of the control groups 1 to 8 and the blank control group were both Yellow means that it does not contain Salmonella Enteritidis; the color of the experimental group turns gr...
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