Kit test method of beta-lactam antibiotics residue in milk

A technology for lactams and detection methods, which is applied in the field of detection kits for β-lactam antibiotic residues in milk, can solve the problems of long detection time, cumbersome detection operations, expensive detection instruments, etc., and achieve the effect of low cost

Active Publication Date: 2010-01-27
GUANGDONG YANTANG DAIRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Wang Zhiqiang et al. "Rapid Detection of Multiple Antibiotic Residues in Fresh Milk by Microbial Inhibition Method" (Chinese Journal of Food Hygiene, 2008, 20(2): 139-141) did not make a kit, the detection time was more than 24 hours, and the operation steps were complicated
[0004] Above-mentioned detection method all is to adopt microbiological detection method, generally exists due to the low (10 5 ~10 6 cfu/ml) and h...

Method used

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  • Kit test method of beta-lactam antibiotics residue in milk
  • Kit test method of beta-lactam antibiotics residue in milk
  • Kit test method of beta-lactam antibiotics residue in milk

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Experimental program
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Effect test

Embodiment 1

[0032]In the first step, Bacillus stearothermophilus ATCC 10208 (Bacillus Stearothermophilus 10208, referred to as BS10208, preserved by China Industrial Microorganism Culture Collection Management Center, address: No. 32, Xiaoyun Road, Chaoyang District, Beijing, zip code: 100027) (also can be Bacillus stearothermophilus Bacillus stearothermis ATCC 10143) was placed in broth liquid medium, and at 50° C., cultivated for 30 h at 150 rpm to make a mother starter; in parts by weight, the broth medium formula was: peptone 10 g, beef extract Powder 25g, sodium chloride 5.0g, distilled water 960g, stir and dissolve evenly, sterilize with 0.08MPa for 20min, cool to 50℃ for later use;

[0033] In the second step, the mother starter is inserted into the production medium at a volume ratio of 3: 100, and cultivated in a fermenter at 55° C. During the cultivation process, the pH is adjusted to 7.5 with a 1.0mol / L NaOH solution. The stirring speed is 120rpm, and it is collected after cult...

Embodiment 2

[0042] The first step is to put Bacillus stearothermophilus ATCC 10208 (Bacillus Stearothermophilus 10208, referred to as BS10208, preserved in China Industrial Microorganism Culture Collection Management Center, address: No. 32 Xiaoyun Road, Chaoyang District, Beijing, postcode: 100027) in the broth liquid culture medium at 45°C and 100 rpm for 24 hours to make a mother starter; in parts by weight, the formula of the broth medium is: peptone 13g, beef extract powder 25g, sodium chloride 4.0g, distilled water 958g; Stir and dissolve evenly, sterilize with 0.10MPa for 15min, cool to 45℃ for later use;

[0043] In the second step, the mother starter is inserted into the production medium at a volume ratio of 1:100, and cultivated in a fermenter at 50° C. During the cultivation process, the pH is adjusted to 8.0 with a 1.5mol / L NaOH solution. The stirring speed is 100rpm, and it is collected after culturing for 24 hours to obtain a suspension of fermented bacteria; in parts by we...

Embodiment 3

[0049] The first step is to put Bacillus stearothermophilus ATCC 10208 (Bacillus Stearothermophilus 10208, referred to as BS10208, preserved in China Industrial Microorganism Culture Collection Management Center, address: No. 32 Xiaoyun Road, Chaoyang District, Beijing, postcode: 100027) in the broth liquid The culture medium was cultured at 55°C and 125rpm for 36h to make a mother starter; in parts by weight, the formula of the broth medium was: peptone 15g, beef extract powder 35g, sodium chloride 6.0g, distilled water 944g; Stir and dissolve evenly, sterilize with 0.09MPa for 18min, cool to 55℃ for later use;

[0050] In the second step, the mother starter is inserted into the production medium at a volume ratio of 5: 100, and cultivated in a fermenter at 65° C. During the cultivation process, the pH is adjusted to 7.0 with a 2.0mol / L NaOH solution. The stirring speed is 150rpm, and it is collected after cultivating for 36 hours to obtain a suspension of fermented bacteria;...

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Abstract

The invention discloses a kit test method of beta-lactam antibiotics residue in milk, comprising the following steps: performing once activation to bacillus stearothermophilus, then culturing the activated bacillus stearothermophilus in a production culture medium at 50-65 DEG C for 24-36h while stirring in a rotation speed of 100-150rpm, centrifugalizing to collect bacteria mire, adding agarose bromcresol purple solution, mixing evenly to ensure the number of spore in each kit reach 1.00*10<7>-2.50*10<8>cfu, subpackaging the spore in porous reaction vessels at 60-70 DEG C, covering aluminum-plastic film, placing upside down, sealing, placing the vessels in aluminum-plastic packages and storing in dark place while keeping from wet. The kit has the advantage of easy operation, high sensibility, good repeatability, low cost and high throughput, can be stored at 4-8 DEG C for 12-18 months and can be used to test beta-lactam antibiotics residue in milk.

Description

technical field [0001] The invention relates to the detection of antibiotic residues, in particular to a detection method of a kit for detecting β-lactam antibiotic residues in milk. Background technique [0002] In the dairy farming industry, β-lactam ring antibiotics are often used to treat mastitis in dairy cows, especially penicillin G is used in large quantities because of its broad spectrum and low cost, resulting in high concentrations of residues in milk. And penicillins can also cause very severe allergic reactions in some individuals. Long-term intake of milk containing such antibiotics will inhibit the normal flora in the human intestinal tract and cause people to develop drug resistance. In many countries, the management of β-lactam drugs in milk, especially penicillin G residues, is very strict. The limit of penicillin G in milk recommended by the World Health Organization has been increased from 0.05 μg / ml (50ppb) in 1976 to the current 0.004 μg / ml (4ppb), th...

Claims

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Application Information

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IPC IPC(8): C12Q1/18C12R1/07
Inventor 刘冬梅余以刚梁洁英吴晖吴祖星陈子剑李晓凤唐语谦
Owner GUANGDONG YANTANG DAIRY
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