Monocyte Listeria monocytogenes strain and application thereof
A monocytogenes, Listeria technology, applied in the field of bacteriology, can solve the problem of lack of strains, etc.
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Embodiment 1
[0024] The isolation and identification of embodiment 1 bacterial strain of the present invention
[0025] 1. Test materials
[0026] 1 Sample source Fresh pork was collected from major supermarkets and bazaars in Harbin.
[0027] 2 The standard strain Listeria monocytogenes CMCC54006 was purchased from China Medical Bacteria Collection Center.
[0028] 3 medium LB 1 , LB 2 Bacteria enrichment solution and SIM power semi-solid medium were purchased from Beijing Luqiao Biological Company, Komagalister chromogenic medium was purchased from Zhengzhou Bosai Bioengineering Co., Ltd., TSA-YE solid medium, TSB-YE liquid culture The base was purchased from Qingdao Haibo Biotechnology Co., Ltd., and the blood plate was made by the bacterial laboratory of Harbin Veterinary Research Institute.
[0029] 4 Primers Listeria monocytogenes-specific primers (primers for detecting the hemolysin gene hlyf) were synthesized by Jinsite Technology (Nanjing) Co., Ltd. The primer sequences are a...
Embodiment 2
[0042] The drug resistance spectrum analysis of embodiment 2 bacterial strains of the present invention
[0043] 1. Test materials
[0044] All the test drug-sensitive tablets were purchased from Beijing Tiantan Drug Biotechnology Development Company.
[0045] 2. Test method
[0046] The D38 strain isolated in Example 1 of the present invention was inoculated into TSB-YE medium, cultured at 37°C for 18 hours, washed with sterilized physiological saline, and the bacterial solution was diluted to 0.5 McFarland turbidity, and the concentration of the bacterial solution was corrected in The inoculation was completed within 15 minutes. Use a sterilized cotton swab to dip in the bacterial solution, rotate and squeeze out the excess bacterial solution on the inner wall of the tube, and then evenly spread and inoculate on the TSA-YE plate for 3 times, rotate the plate 60° each time, and finally spread a circle along the edge of the plate. Put the cover on and let it sit for 5 minut...
Embodiment 3
[0050] Embodiment 3 Determination of the tetracycline drug-resistant gene of bacterial strain of the present invention
[0051] 1. Test materials
[0052] 1 Reagents: DNA Marker and Ex Taq enzymes were purchased from Dalian Bao Biological Engineering Co., Ltd.
[0053] 2 Primers: TetM-specific primers for the tetracycline resistance gene were synthesized by Jinsite Technology (Nanjing) Co., Ltd. The primer sequences are as follows:
[0054] tetm-U 5′-ACAGAAAGCTTTATTATAAC-3′
[0055] tetm-L 5′-TGGCGTGTCTATGATGTTCAC-3′
[0056] 2. Test method
[0057] The D38 strain of the present invention was inoculated in TSB-YE medium, and after culturing at 37° C. for 24 hours, the genome of the strain was extracted with a genomic DNA extraction kit. Then the tetracycline resistance gene tetM was amplified by PCR under the reaction conditions of 95°C pre-denaturation for 5 min, 94° denaturation for 30 s, 50° annealing for 30 s, 72° extension for 1 min, 30 cycles, and 72° final extensio...
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