Rapid detection kit for drug susceptibility of mycoplasma bovis and preparation method of rapid detection kit

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

Active Publication Date: 2018-06-19
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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  • Rapid detection kit for drug susceptibility of mycoplasma bovis and preparation method of rapid detection kit
  • Rapid detection kit for drug susceptibility of mycoplasma bovis and preparation method of rapid detection kit
  • Rapid detection kit for drug susceptibility of mycoplasma bovis and preparation method of rapid detection kit

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Experimental program
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preparation example Construction

[0029] Second aspect, the present invention provides a kind of preparation method of described Mycoplasma bovis drug susceptibility rapid detection kit, comprises the following steps:

[0030] (1) Preparation of Mycoplasma bovis culture medium: dissolve 12g of PPLO powder, 3g of short peptide, 300mg of morroniside, 4g of yeast powder, 2.5g of glucose, and 3.5g of sodium pyruvate in 750mL of ddH2O, add 100mL of MEM medium, 1% (w / v) phenol red solution 2mL, after sterilizing at 116°C for 20min, add 150mL of horse serum inactivated at 56°C and 300,000 units of penicillin, adjust the pH value to 7.6-7.8, dispense into 15mL culture tubes, 9ml / Tube.

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

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

[0033] S2. Dissolve erythromycin, florfenicol, norfloxacin, kanamycin, tilmicosin, telamycin, garamimycin and tetracycline in 1-5mL buffer, and dilute to 128μg...

Embodiment 1

[0036] Embodiment 1 a kind of mycoplasma bovis culture fluid

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

[0040] The Mycoplasma bovis culture solution is composed of the following components and contents: PPLO powder 10-15g, short peptide 2-4g, morroniside 100-500mg, yeast powder 2-6g, glucose 1-3g, sodium pyruvate 1.5-4.5g , MEM medium 80-120 mL, 1% (w / v) phenol red solution 2 mL, horse serum 100-300 mL, 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 thereof, in particular to a rapid detection kit for drug sensitivity of Mycoplasma bovis and a preparation method thereof. Background technique [0002] Mycoplasma bovis (mycoplasma bovis) is one of the important pathogens that cause mastitis, calf pneumonia and arthritis in adult cows. It is highly contagious and often mixed with other pathogenic bacteria, seriously threatening 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 pneumonia in my country in 2008, there have been cases of mycoplasma pneumonia deaths in calves and cases of mycoplasma mastitis in adult cows in cattle farms. . The incidence of the disease is high, improper treatment or death of calves, often mixed with other bacteria or viruses, the clinical symptoms are complex and diverse, so ...

Claims

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Application Information

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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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