LAMP (loop-mediated isothermal amplification) kit for rapid detection of staphylococcus aureus
A staphylococcus, aureus technology, applied in the direction of microorganism-based methods, microbial determination/testing, microorganisms, etc., can solve the problems of low sensitivity, poor specificity, time-consuming and labor-intensive, etc., and achieve high sensitivity, simple steps, and repeatability high effect
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Embodiment 1
[0077] 1. The composition and preparation of the new rapid visual detection of Staphylococcus aureus LAMP kit, reagent composition:
[0078] a) LAMP reaction mixture
[0079] The LAMP reaction mixture consists of 2.5 μl of LAMP 10×buffer, 2.0 μl of 10 μmol / L internal primers FIP and BIP, 1.0 μl of 10 μmol / L external primers F3 and B3, 3.0 μl of 10 mmol / L dNTPs, 50 mmol / LMgSO 4 3.0 μl, 5.0mol / L betaine, 4.5 μl, Bst DNA polymerase large fragment 1.0 μl, the total volume of the reaction solution is 20 μl.
[0080] Wherein the primer sequence is as follows:
[0081] Upstream outer primer F3: 5'-GCGATTGATGGTGATACGGTTA-3';
[0082] Downstream outer primer B3: 5′-GCCACGTCCATATTTATCAGTTCT-3′;
[0083] Upstream internal primer FIP: 5′-GGATGCTTTGTTTCAGGTGTATCATGTACAAAGGTCAACCAATGAC-3′;
[0084] Downstream internal primer BIP: 5′-TATGGTCCTGAAGCAAGTGCAGACCTTTGTCAAACTCGACTTC-3′;
[0085] b) Standard positive template
[0086] The standard positive template pMD18-T-nuc is a pMD18-T-n...
Embodiment 2
[0096] 1. The composition and preparation of the new rapid visual detection of Staphylococcus aureus LAMP kit, the difference from the kit described in Example 1 is that the specific primer sequences are different. The primer sequences of this kit are as follows:
[0097] Upstream outer primer F3: 5′-ACATAAAGAACCTGCGACA-3′;
[0098] Downstream outer primer B3: 5'-CTTTGTCAAACTCGACTTCA-3';
[0099] Upstream internal primer FIP: 5′-CTGAATGTCATTGGTTGACCTTTGTAATTAAAGCGATTGATGGTGAT-3′;
[0100] Downstream inner primer BIP:
[0101] 5′-CACCTGAAACAAAGCATCCTAAAAAGCATTTTCTACCATCTTTTTCGTA-3′
[0102] 2. The method for detecting Staphylococcus aureus using the PCR kit to detect Staphylococcus aureus in industrial food rapidly based on the loop-mediated isothermal amplification technology is exactly the same as the method described in Example 1, and the repeated test was repeated 3 times, and the obtained The detection results are the same, the experimental group has precipitation, and ...
Embodiment 3
[0104] 1. The composition and preparation of the new rapid visual detection of Staphylococcus aureus LAMP kit, the difference from the kit described in Example 1 is that the specific primer sequences are different. The primer sequences of this kit are as follows:
[0105] Upstream outer primer F3: 5'-GCGACATTAATTAAAGCGATTG-3';
[0106] Downstream outer primer B3: 5'-CTTTGTCAAACTCGACTTCAA-3';
[0107] Upstream inner primer FIP:
[0108] 5'-GCTTTGTTTCAGGTGTATCAACCAAATTAATGTACAAAGGTCAACCAATG-3';
[0109] Downstream internal primer BIP: 5′-AAGGTGTAGAGAAATATGGTCCTGATTCTTTGCATTTTCTACCATCT-3′;
[0110] 2. The method for detecting Staphylococcus aureus using the PCR kit to detect Staphylococcus aureus in industrial food rapidly based on the loop-mediated isothermal amplification technology is exactly the same as the method described in Example 1, and the repeated test was repeated 3 times, and the obtained The detection results are the same, the experimental group has precipitation...
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