Cow recessive endometritis early diagnosis kit

A technology for endometritis and early diagnosis, which is applied in the field of immunological testing and can solve the problems of difficult control of semen temperature, difficult operation, and difficult sample collection.

Inactive Publication Date: 2011-05-04
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

шипилo B et al proposed that biological diagnosis with semen is easy to operate, but it is not easy to control the temperature of semen
As for the method of reacting urine with silver nitrate used by Γa B p иш, the disadvantage is that the sample is not easy to collect, and the urination of cows is very random, so it is not easy to operate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: a kind of milk cow recessive endometritis early diagnosis kit of the present invention, it is to utilize the standard antibody of SAA, IL-6 and IL-8 to coat the microtiter plate, make competing antibody with novel monoclonal antibody, Use horseradish peroxidase to label the secondary antibody, prepare serum dilution solution, washing solution, substrate solution A, substrate solution B, H 2 o 2 , stop solution, assembly composition;

[0043] 1. Preparation of SAA, IL-6 and IL-8 antibodies

[0044] (1) Antigen acquisition and animal vaccination

[0045] Prokaryotic culture of E.coli, separated and purified by Ni column, and then multi-point intradermal injection of antigen emulsified with Freund's complete adjuvant on the back of the rabbit, each point needs to inject about 0.1ml, a total of 0.5ml injection, 2 Weekly booster immunization;

[0046] (2) Collection and storage of antiserum: Blood is collected from the ear vein or ear artery, the serum is s...

Embodiment 2

[0071] Embodiment 2: The following explanations are made on the relevant issues of the recessive endometritis competition ELISA detection kit:

[0072] 1 About the collection and preservation of specimens

[0073]Most ELISA tests use serum as a sample. In addition to still containing fibrinogen and anticoagulant in plasma, other components are equal to serum. Except for special circumstances, serum is used as the test specimen in medical tests. Serum and plasma can be used equally in ELISA. Serum samples can be collected according to conventional methods, and attention should be paid to avoid hemolysis. Substances with peroxidase activity will be released when red blood cells are lysed. In the ELISA assay using HRP as a marker, hemolysis samples may increase non-specific color development.

[0074] Serum samples should be tested when they are fresh. If there is bacterial contamination, the bacteria may contain endogenous HRP, which will also produce false positive reaction...

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PUM

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Abstract

The invention provides a quick and accurate cow recessive endometritis early diagnosis kit. The kit is assembled by serum amyloid A (SAA), interleukin-6 (IL-6) and IL-8 standard antibody enveloped ELISA plates, a novel monoclonal antibody serving as a competitive antibody, a horse radish peroxidase labeled secondary antibody and the like, and can be prepared by preparing SAA, IL-6 and IL-8 antibodies, the horse radish peroxidase labeled secondary antibody, an enzyme-linked immuno sorbent assay (ELISA) reagent and an ELISA plate. In the kit, an ELISA method is mainly adopted. The ELISA method has a wide application range and high operation speed and can eliminate certain vaccine reactions; by the method, a result can be acquired in two minutes; and the kit can be used for laboratory detection, pasture operation and field operation and is suitable for clinical detection and farmer on-site detection.

Description

(1) Technical field [0001] The invention relates to an immunological testing technology, in particular to a kit for early diagnosis of dairy cow latent endometritis. (2) Background technology [0002] Since G.Widal and A.Sicad used agglutination reaction to diagnose typhoid fever in 1896, immunology and medical examination have formed an indissoluble bond, and it has been a century. With the development of immunological technology, immunological examination has become an important part of veterinary clinical examination. [0003] The detection objects of immunological tests are substances with immune activity, including detection methods and clinical significance. Immunological tests can be divided into two categories: cellular immune tests and humoral immune tests. The detection of immune active cells and their functions belongs to the former, and the detection of antigens, antibodies, complements, etc. belongs to the latter. [0004] In recent years, the rapid developmen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/535
Inventor 李德军刘运枫王海彬
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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