CN104198658A - Method for carrying out laboratory evaluation on veterinary medicines or disinfectants for cultivation - Google Patents

Abstract

The invention discloses a method for carrying out laboratory evaluation on veterinary medicines or disinfectants for cultivation. The method comprises the following steps: (1) preparing a loading bacterium solution; (2) preparing disinfectants of different concentrations; (3) loading and incubating; and (4) judging the result: a, comparing the minimal inhibitory concentration (MIC), b, comparing the OD450 bacteriostatic trend, and c, comparing effective bacteriostatic concentration and cost; finally, carrying out comprehensive appraisal by combining with the MIC, the OD450 bacteriostatic trend and the effective bacteriostatic concentration and cost, and selecting the optimal disinfectant. Laboratory evaluation is carried out on the veterinary medicines or disinfectants of farms for preventing and treating diseases by adopting a trace drug sensitive test, and a certain reference is provided for correct use of the medicines.

Description

A kind of method that cultivates veterinary drug used or sanitizer and carry out laboratory evaluation

Technical field

The present invention relates to a kind of detection method that cultivates veterinary drug used or sanitizer, especially a kind of method that cultivates veterinary drug used or sanitizer and carry out laboratory evaluation.

Background technology

At present, before buying market veterinary drug and disinfectant, can evaluate out efficiently or for this plant active drug, also can provide effective veterinary drug to Prevention and cure of epidemic situation simultaneously by micro-susceptibility.

Summary of the invention

The object of this invention is to provide a kind of method that cultivates veterinary drug used or sanitizer and carry out laboratory evaluation.

This cultivation veterinary drug used or sanitizer carry out the method for laboratory evaluation:

1. loading bacterium solution preparation:

By separating Escherichia coli in water sample, the single bacterium colony of picking is mixed in the TSB of 5mL, 37 DEG C of baking ovens leave standstill hatches 2h, now bacterium liquid muddiness, get 8-10 gradient of 10 times of dilutions of 100 μ L bacterium liquid, evenly be applied in TSA and count after 37 DEG C of standing 24h, be then adjusted to 105CFU/mL according to the bacterial concentration of counting, for loading bacterium liquid;

2. variable concentrations sanitizer preparation:

According to minimum working concentration and the highest working concentration of operation instruction on sanitizer, doubling dilution experiment need comprise minimum and the highest working concentration by concentration, is divided into 6 gradients;

3. loading is hatched:

(1) the 1st front 6 holes of row in 96 microwell plates are added respectively to 6 kinds of variable concentrations sanitizers, 100 μ L, the 7th hole adds loading bacterium liquid 200 μ L as positive control, and the 8th hole adds this sanitizer 200 μ L;

(2) the 1st front 6 holes of row add 100 μ L loading bacterium liquid again, and causing every Kongzui end-body is 200 μ L;

(3) the 2nd rows are as the parallel control of the 1st row's sanitizer, and other sanitizers also, with after aforesaid way application of sample, are built plate lid; Then be placed in 37 DEG C, leave standstill and hatch, respectively get 4 points respectively at 3-6h and 12-24h and observe MIC and measure OD450 value;

4. result is judged:

(1) MIC minimal inhibitory concentration comparison: recommended density or minimum working concentration comparison in the MIC of sanitizer and its operation instruction, if lower than recommended density, sanitizer effect is better, otherwise poor; Different sanitizer MIC comparison, MIC is lower, but must first be less than recommended density, and sanitizer effect is better;

(2) the antibacterial trend comparison of OD450: for the identical sanitizer of MIC value, can pass through the antibacterial trend of OD450, analyze its bacteriostasis time and stability, judge good and bad;

(3) effectively Mlc expense comparison: according to effective Mlc (MIC), more each sanitizer dilutes the same water yield, the expense spending;

Finally, in conjunction with MIC and the antibacterial trend of OD450 and effectively Mlc expense synthetical comparison and assessment, select optimum sanitizer.

The beneficial effect of the invention:

This invention adopts micro-drug sensitive test to carry out laboratory evaluation for plant's disease preventing and treating veterinary drug used or sanitizer, is the certain reference frame of correct use drug provision.

Embodiment

Embodiment:

1, loading bacterium solution preparation

By separating Escherichia coli in water sample, the single bacterium colony of picking is mixed in the TSB of 5mL, 37 DEG C of baking ovens leave standstill hatches 2h (now bacterium liquid muddiness), get 8-10 gradient of 10 times of dilutions of 100 μ L bacterium liquid, evenly be applied in TSA and count after 37 DEG C of standing 24h, then be adjusted to 105CFU/mL according to the bacterial concentration of counting, for loading bacterium liquid.

2, variable concentrations sanitizer preparation

According to minimum working concentration and the highest working concentration of operation instruction on sanitizer, doubling dilution experiment need comprise minimum and the highest working concentration (general 6 gradients) by concentration.

3, loading is hatched

(1) the 1st front 6 holes of row in 96 microwell plates are added respectively to 6 kinds of variable concentrations sanitizers, 100 μ L, the 7th hole adds loading bacterium liquid 200 μ L as positive control, and the 8th hole adds this sanitizer 200 μ L.

(2) the 1st front 6 holes of row add 100 μ L loading bacterium liquid again, and causing every Kongzui end-body is 200 μ L.

(3) the 2nd rows are as the parallel control of the 1st row's sanitizer, and other sanitizers also, with after aforesaid way application of sample, are built plate lid.Then be placed in 37 DEG C, leave standstill and hatch, respectively get 4 points respectively at 3-6h and 12-24h and observe MIC and measure OD450 value.

4, result is judged:

(1) MIC (minimal inhibitory concentration) relatively: relatively, if lower than recommended density, sanitizer effect is better, otherwise poor for recommended density in the MIC of sanitizer and its operation instruction (or minimum working concentration); Different sanitizer MIC comparison, MIC lower (but must first be less than recommended density), sanitizer effect is better.

(2) the antibacterial trend comparison of OD450: for the identical sanitizer of MIC value, can pass through the antibacterial trend of OD450, analyze its bacteriostasis time and stability, judge good and bad.

(3) effectively Mlc expense comparison: according to effective Mlc (MIC), more each sanitizer dilutes the same water yield, the expense spending.

Finally, in conjunction with MIC and the antibacterial trend of OD450 and effectively Mlc expense synthetical comparison and assessment, select optimum sanitizer.

Claims (1)

1. cultivation veterinary drug used or sanitizer carry out a method for laboratory evaluation, it is characterized in that:

1. loading bacterium solution preparation:

By separating Escherichia coli in water sample, the single bacterium colony of picking is mixed in the TSB of 5mL, 37 DEG C of baking ovens leave standstill hatches 2h, now bacterium liquid muddiness, get 8-10 gradient of 10 times of dilutions of 100 μ L bacterium liquid, evenly be applied in TSA and count after 37 DEG C of standing 24h, be then adjusted to 105CFU/mL according to the bacterial concentration of counting, for loading bacterium liquid;

2. variable concentrations sanitizer preparation:

According to minimum working concentration and the highest working concentration of operation instruction on sanitizer, doubling dilution experiment need comprise minimum and the highest working concentration by concentration, is divided into 6 gradients;

3. loading is hatched:

(1) the 1st front 6 holes of row in 96 microwell plates are added respectively to 6 kinds of variable concentrations sanitizers, 100 μ L, the 7th hole adds loading bacterium liquid 200 μ L as positive control, and the 8th hole adds this sanitizer 200 μ L;

(2) the 1st front 6 holes of row add 100 μ L loading bacterium liquid again, and causing every Kongzui end-body is 200 μ L;

(3) the 2nd rows are as the parallel control of the 1st row's sanitizer, and other sanitizers also, with after aforesaid way application of sample, are built plate lid; Then be placed in 37 DEG C, leave standstill and hatch, respectively get 4 points respectively at 3-6h and 12-24h and observe MIC and measure OD450 value;

4. result is judged:

(1) MIC minimal inhibitory concentration comparison: recommended density or minimum working concentration comparison in the MIC of sanitizer and its operation instruction, if lower than recommended density, sanitizer effect is better, otherwise poor; Different sanitizer MIC comparison, MIC is lower, but must first be less than recommended density, and sanitizer effect is better;

(2) the antibacterial trend comparison of OD450: for the identical sanitizer of MIC value, can pass through the antibacterial trend of OD450, analyze its bacteriostasis time and stability, judge good and bad;

(3) effectively Mlc expense comparison: according to effective Mlc (MIC), more each sanitizer dilutes the same water yield, the expense spending;

Finally, in conjunction with MIC and the antibacterial trend of OD450 and effectively Mlc expense synthetical comparison and assessment, select optimum sanitizer.