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Method for detecting tylosin and special enzyme-linked immune reagent kit thereof

An enzyme-linked immunosorbent reagent, tylosin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inappropriate screening of a large number of samples, sensitivity and specificity limitations, cumbersome processing and determination operations, etc., to achieve The inspection method is convenient and easy, the pretreatment process is simple, and the effect of shortening the detection time

Inactive Publication Date: 2009-08-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the microbial detection method is economical and easy to operate, its sensitivity and specificity are limited when there are other microbial inhibitors in the sample; High sensitivity, but cumbersome sample pretreatment and determination operations, high cost, not suitable for screening a large number of samples, can be used as a confirmatory analysis of residues

Method used

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  • Method for detecting tylosin and special enzyme-linked immune reagent kit thereof
  • Method for detecting tylosin and special enzyme-linked immune reagent kit thereof
  • Method for detecting tylosin and special enzyme-linked immune reagent kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Preparation and detection method of an enzyme-linked immunosorbent assay kit using a conjugate of a tylosin hapten and a carrier protein as a coating source

[0035]The ELISA kit using the conjugate of tylosin hapten and carrier protein as the coating source includes:

[0036] (1) A microtiter plate coated with a conjugate of tylosin and a carrier protein;

[0037] (2) Alkaline phosphatase-labeled goat anti-mouse anti-antibody working solution: Dilute the alkaline phosphatase-labeled goat anti-mouse anti-antibody to a protein concentration of 0.1-1 μg / L with enzyme-labeled antibody diluent. The alkaline phosphatase-labeled anti-antibody, the enzyme-labeled antibody diluent is a solution containing 0.1‰ (mass concentration) glycerin and 1% thimerosal preservative. 12ml / bottle, 1 bottle.

[0038] (3) Tylosin standard solution: 6 bottles of tylosin series standard solution, 0μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L, 40.5μg / L, 121.5μg / L, 1~3ml / bottle. The tylosin dru...

Embodiment 2

[0082] Example 2, ELISA kit using anti-antibody as coating source and its preparation method

[0083] ELISA kits with tylosin anti-antibody as coating source include:

[0084] (1) Enzyme plate coated with tylosin anti-antibody;

[0085] (2) Alkaline phosphatase-labeled tylosin hapten working solution: Dilute the alkaline phosphatase-labeled tylosin hapten to a protein concentration of 0.1-1 μg / L with enzyme-labeled hapten diluent . The enzyme-labeled hapten diluent is a solution containing 50% glycerin and 1% thimerosal preservative. 12ml / bottle, 1 bottle.

[0086] (3) Tylosin standard solution: 6 bottles of tylosin series standard solution, 0μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L, 40.5μg / L, 121.5μg / L, 1~3ml / bottle. The tylosin medicine diluent used is a phosphate buffer solution containing 5% (mass concentration) N,N'-dimethylformamide (DMF) and 1% ovalbumin (OVA).

[0087] (4) Chromogenic agent: 4-nitrophenol phosphate buffer. Both are 8ml / bottle.

[0088] (5) Tylosin rabb...

Embodiment 3

[0111] Embodiment 3, test kit precision, accuracy and shelf life test

[0112] 1. Kit precision test

[0113] (1) Precision test of standard product

[0114] Three batches of the kits prepared in Example 1 and Example 2 were taken respectively for precision experiments, and 10 kits were extracted from each batch of kits, and 20 microwells were respectively extracted from the enzyme-linked plate of each kit. Measure the absorbance value (OD value) of the 13.5 μg / L standard solution, and calculate the coefficient of variation. The measurement results of the three batches of kits in Example 1 are shown in Table 1, and the results show that the coefficient of variation ranges from 4.2% to 10.2%.

[0115] The kit standard repeatability test of table 1 embodiment 1

[0116]

[0117] The measurement results of the three batches of kits in Example 2 are shown in Table 2, and the results show that the coefficient of variation ranges from 3.7% to 8.6%.

[0118] Table 2. The kit s...

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Abstract

The invention discloses a method for detecting tylosin and a special ELISA kit. The ELISA kit for detecting tylosin includes tylosin-specific antibodies, a coating source and an enzyme marker; the coating source is a conjugate of a tylosin hapten and a carrier protein or Anti-antibody; the enzyme label is an enzyme-labeled anti-antibody or an enzyme-labeled tylosin hapten; when the coating is originally a conjugate of a tylosin hapten and a carrier protein, the enzyme label It is an enzyme-labeled anti-antibody; when the coating is originally an anti-antibody, the enzyme label is an enzyme-labeled tylosin hapten; the method of the present invention is easy to operate, low in cost, high in sensitivity, capable of on-site monitoring and suitable for A large number of samples are screened to detect the residues of tylosin in animal tissues (muscle, liver) and other samples.

Description

technical field [0001] The invention relates to a method for detecting tylosin and a special ELISA kit. Background technique [0002] The trade name of Halofuginone is Stenorol. It is a 0.6% premix of tylosin hydrobromide. It has special effects and has no cross-resistance with other coccidiostats, so it is often added to feed to control animal diseases caused by coccidia. It is a drug additive widely used in feed at present. However, tylosin is highly toxic to aquatic animals (such as fish and shrimp), and can harm humans through the food chain. If a person eats poultry tissue with a high residual concentration of tylosin, he or she will experience physical discomfort and other poisoning phenomena. Announcement No. 235 of the Ministry of Agriculture of the People's Republic of China "Maximum Residue Limits of Veterinary Drugs in Animal Foods" stipulates that the maximum residue limit of tylosin in chicken liver tissue is 130 μg / kg. [0003] At present, the methods common...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/577G01N33/535
Inventor 沈建忠何方洋万宇平史为民冯才伟张素霞吴小平汪善良
Owner CHINA AGRI UNIV
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