Method for making gentamicin sulphate standard detection curve and gentamicin sulphate enzyme linked fast detection method

A technology of gentamicin sulfate and detection standards, which is applied in the direction of measuring devices, biological testing, material inspection products, etc., and can solve the problems of long detection time of detection methods

Active Publication Date: 2007-03-14
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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Problems solved by technology

[0003] The purpose of the present invention is to solve the problem that the detection time of current gentamicin sulfate detection method is long, and provide a kind of method of making gentamicin sulfate detection standard curve and gentamicin sulfate ELISA rapid detection method

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  • Method for making gentamicin sulphate standard detection curve and gentamicin sulphate enzyme linked fast detection method

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specific Embodiment approach 1

[0005] Specific embodiment one: the method for making gentamicin sulfate detection standard curve in this embodiment is as follows: (1) get 0.17g gentamicin sulfate and dissolve in 20mL, 0.12g bovine serum albumin and 0.12g ovalbumin respectively. Phosphate buffer with a pH value of 6.8, then add 10 mL of glutaraldehyde with a concentration of 2.5%, and stir for 28 to 32 hours; (2) dialyze in a phosphate buffer with a pH value of 7.0 for 6 to 8 hours, and the Collect the antigen gentamicin sulfate protein conjugate; (3) take 2mL gentamicin sulfate bovine serum albumin conjugate and emulsify with 8mL complete Freund's adjuvant or 8mL incomplete Freund's adjuvant, each mouse 0.2 mL of emulsified antigen was injected internally, and gentamicin sulfate was injected for booster immunization until the antibody titer was above 1:16; (4) the serum was separated; (5) the antibody in the serum was separated by ammonium sulfate salting out method, and then the antibody in the serum was se...

specific Embodiment approach 2

[0006] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the volume ratio of the antigen diluent to the antibody diluent in step (8) is 1:1-10. Other steps are the same as those in Embodiment 1.

specific Embodiment approach 3

[0007] Embodiment 3: The difference between this embodiment and Embodiment 1 is that in step (5), a saturated ammonium sulfate three-step method is used to separate antibodies. Other steps are the same as those in Embodiment 1.

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Abstract

The related method for gentamycin sulfate detection standard curve comprises: coupling protein and gentamycin sulfate to inject into rat body and prepare antibody, selecting the work concentration for antigen and antibody reaction by chess board method, and selecting holes with OD450 value as 1-1.5 to calculate and draw the curve. The enzyme linked immunity fast detection method comprises: 1. diluting antibody, adding gradient-diluting sample, enzyme link reacting for 1-1.5h at 41Deg, developing with TMB; 2. selecting the sample with 0.4-1.5OD value; 3. checking data in curve, and multiplying dilution time to obtain the residual gentamycin sulfate. This detection method needs time only 1-1.5h.

Description

technical field [0001] The invention relates to a method for preparing a gentamicin sulfate detection standard curve and a gentamicin sulfate detection method. Background technique [0002] As a commonly used antibiotic, gentamicin sulfate is widely used in the prevention and control of agricultural, livestock, poultry, forestry and aquatic products. Due to the abuse of users, the problem of large residues of gentamicin sulfate often occurs. Residues of gentamicin sulfate will seriously endanger human health and the export of domestic livestock and poultry products and aquatic products, so strict safety inspections must be carried out on food. At present, there are chromatographic detection methods and microbial detection methods for the detection of gentamicin sulfate, but there are problems of long detection time, and the sample pretreatment operation of chromatographic detection method is difficult, cannot detect trace residues, and cannot be quantitatively analyzed. Fac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/543
Inventor 王静曹维强丁志刚金芬邵华杨锚
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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