A kind of mycoplasma bovis drug susceptibility rapid detection kit and preparation method thereof

A detection kit and technology for Mycoplasma bovis, applied in biochemical equipment and methods, microorganism-based methods, and microbial determination/inspection, etc., can solve the problems of no commercialized kits, no national standards or patents, etc. Shorter color development time, high efficiency, and strong specificity

Active Publication Date: 2020-05-22
DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there is no national standard or patent to refer to in terms of drug susceptibility testing of Mycoplasma bovis, and there is no commercial kit yet.

Method used

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  • A kind of mycoplasma bovis drug susceptibility rapid detection kit and preparation method thereof
  • A kind of mycoplasma bovis drug susceptibility rapid detection kit and preparation method thereof
  • A kind of mycoplasma bovis drug susceptibility rapid detection kit and preparation method thereof

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preparation example Construction

[0029] In a second aspect, the present invention provides a preparation method of the Mycoplasma bovis drug susceptibility rapid detection kit, comprising the following steps:

[0030] (1) Preparation of Mycoplasma bovis culture solution: dissolve 12 g of PPLO powder, 3 g of short peptide, 300 mg of morroniside, 4 g of yeast powder, 2.5 g of glucose, and 3.5 g of sodium pyruvate in 750 mL of ddH 2 O, add 100 mL of MEM medium, 2 mL of 1% (w / v) phenol red solution, sterilize at 116 °C for 20 min, add 150 mL of horse serum inactivated at 56 °C and 300,000 units of penicillin, and adjust the pH to 7.6 to 7.8 , dispensed into 15mL culture tubes, 9mL / tube.

[0031] (2) Preparation of drug sensitive plate:

[0032] S1. The 96-well plate was washed roughly, finely washed, and dried at a constant temperature of 40°C for use;

[0033] S2. Dissolve erythromycin, florfenicol, norfloxacin, kanamycin, tilmicosin, teramycin, gamycin and tetracycline into 1-5 mL buffer, and dilute to 128μg...

Embodiment 1

[0036] Embodiment 1 A kind of Mycoplasma bovis culture solution

[0037] The Mycoplasma bovis culture solution is composed of the following components and contents: 10-15 g of PPLO powder, 2-4 g of short peptide, 100-500 mg of morroniside, 2-6 g of yeast powder, 1-3 g of glucose, and 1.5-4.5 g of sodium pyruvate , MEM medium 80~120mL, 1% (w / v) phenol red solution 2mL, horse serum 100~300mL, 300,000 units of penicillin and ddH 2 O 700~800mL.

[0038] The peptide chain is an oligomeric short peptide of Tyr-Leu-Tyr-Glu-Val-Ala.

Embodiment 2

[0039] Embodiment 2 A kind of Mycoplasma bovis culture solution

[0040] The Mycoplasma bovis culture solution is composed of the following components and contents: 10-15 g of PPLO powder, 2-4 g of short peptide, 100-500 mg of morroniside, 2-6 g of yeast powder, 1-3 g of glucose, and 1.5-4.5 g of sodium pyruvate , MEM medium 80~120mL, 1% (w / v) phenol red solution 2mL, horse serum 100~300mL, 300,000 units of penicillin and ddH 2 O 700~800mL.

[0041] The peptide chain is an oligomeric short peptide of Tyr-Leu-Tyr-Glu-Val-Ala.

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Abstract

The invention relates to an in vitro diagnostic reagent and a preparation method thereof and in particular relates to a rapid detection kit for drug susceptibility of mycoplasma bovis and a preparation method of the rapid detection kit. The invention provides the rapid detection kit for the drug susceptibility of the mycoplasma bovis; the kit comprises a mycoplasma culture solution, a drug susceptibility plate and an antibiotic drug. The mycoplasma culture solution is prepared from the following components according to the content: 10 to 15g of PPLO (Pleuropneumonia-like Organism) powder, 2 to4g of oligopeptide, 100 to 500mg of morroniside, 2 to 6g of yeast powder, 1 to 3g of glucose, 1.5 to 4.5g of sodium pyruvate, 80 to 120mL of an MEM culture medium, 2mL of a 1 percent (w / v) phenol redsolution, 100 to 300mL of horse serum, 0.3 million units of penicillin and 700 to 800mL of ddH2O. The culture solution provided by the invention can be used for rapidly promoting the growth of the mycoplasma bovis and detection can be finished within 20 to 24h; the rapid detection kit has stable performance and strong specificity.

Description

technical field [0001] The invention relates to an in vitro diagnostic reagent and a preparation method, in particular to a rapid detection kit for drug sensitivity of Mycoplasma bovis and a preparation method thereof. Background technique [0002] Mycoplasma bovis is one of the important pathogens causing mastitis in adult cows, pneumonia in calves and arthritis. It is highly contagious and often mixed with other pathogenic bacteria, which seriously threatens the health of cattle. caused huge economic losses. Mycoplasma bovis was first isolated from the milk of cows with mastitis in 1961. Since the first report of Mycoplasma bovis pneumonia in my country in 2008, there have been cases of death of Mycoplasma bovis pneumonia in calves and cases of Mycoplasma bovis mastitis in adult cows. . The disease has a high incidence, improper treatment of calves or causes death, and the clinical symptoms are often complex and diverse when mixed with other bacteria or viruses. Therefore...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12R1/35
CPCC12Q1/04
Inventor 张亮杨宏军朱曼玲王基隆解晓莉程凯慧苗自利吕亚妮张云飞杨美刘来兴
Owner DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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