PCR detection method of Salmonella typhimurium, nucleic acid and primer pair thereof
A Salmonella and detection method technology, which is applied in the field of Salmonella typhimurium PCR detection method and its nucleic acid and primer pairs, can solve the problem of low specificity of nucleic acid and primers and detection target of the uninvented Salmonella typhimurium serotype PCR detection method. Increase the detection complexity and other issues to achieve the effect of practicality, simple result judgment and short detection time
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Embodiment 1
[0021] Establishment of PCR detection method for serotypes of Salmonella typhimurium
[0022] Step 1, designing primers according to the conserved sequence in the genomic DNA sequence of Salmonella typhimurium
[0023] Find the specific STM4495 gene from the Salmonella typhimurium genome DNA sequence, and use it as the detection target gene of Salmonella typhimurium, the gene sequence is shown in SEQ ID NO: 1;
[0024] Input the DNA sequence of the restriction endonuclease gene into the primer design software Primer Premier 5.0 to design primers, set the GC% range to 40-60%, and the product size range to 100-1000bp, and select primers from the alternative primer pairs , the primer sequences are as follows (primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.):
[0025] STM4-L: 5'-GGTGGCAAGGGAATGAA-3' (SEQ ID NO: 2);
[0026] STM4-R: 5'-CGCAGCGTAAAGCAACT-3' (SEQ ID NO: 3);
[0027] Step 2, DNA template preparation
[0028] Inoculate Salmon...
Embodiment 2
[0041] Detection of suspected strains of Salmonella typhimurium
[0042] Using the PCR detection method for Salmonella typhimurium established in Example 1, 61 suspected strains of Salmonella isolated from food samples were detected. The food samples were collected in suburban supermarkets and bazaars in Shanghai. For sample processing and isolation of suspected strains, refer to the national standard GB / T 4789.4-2008.
[0043] From it, 9 suspected bacterial strains were detected as positive results, and these 9 suspected bacterial strains were identified as O4:Hi:5 by the Salmonella diagnostic serum (Lanzhou Institute of Biological Products), and it was determined that it was Salmonella typhimurium (see the product manual for the serum identification steps and national standard GB / T 4789.4-2008), other suspected strains were identified as not Salmonella typhimurium. This example proves that the PCR detection method for Salmonella typhimurium has very high reliability.
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