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Dairy cow ketosis diagnostic kit and detecting method thereof

The technology of a diagnostic kit and detection method is applied in the direction of material analysis by observing the influence on chemical indicators, and analysis by causing the material to undergo chemical reaction, which can solve the problems of stress response, high detection cost, damage, etc. To achieve the effect of high sensitivity and specificity, good stability and convenient use

Inactive Publication Date: 2014-05-14
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The diagnosis of subclinical ketosis in dairy cows mainly depends on the detection of ketone body content in the blood. In large-scale dairy farms, the workload of collecting cow blood is large, time-consuming and laborious, and the requirements for equipment are high, and the detection cost is relatively high. High, according to my country's actual national conditions, it is still difficult to popularize and use it on a large scale
In addition, blood collection itself is a kind of stress response to dairy cows, and there is a certain amount of damage. For ketosis in dairy cows, normalized and regular monitoring is required. Normalized blood collection will lead to stress reactions in various aspects of dairy cows

Method used

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  • Dairy cow ketosis diagnostic kit and detecting method thereof
  • Dairy cow ketosis diagnostic kit and detecting method thereof
  • Dairy cow ketosis diagnostic kit and detecting method thereof

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Experimental program
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Effect test

Embodiment 1

[0022] Solution preparation and sample detection

[0023] Below in conjunction with embodiment and accompanying drawing, the present invention will be described in further detail:

[0024] (1) The detection principle of this kit

[0025]

[0026] (2) Composition and preparation of the solution

[0027] Reagent 1 is a mixture of diaphorase, nicotinamide adenine dinucleotide and β-hydroxybutyrate dehydrogenase, mixed in a mass ratio of 2:2:1. When preparing, weigh 200mg diaphorase, 200mg nicotinamide adenine dinucleotide and 100mg β-hydroxybutyrate dehydrogenase, mix them evenly and put them in brown plastic, and store them sealed at -20°C for later use.

[0028] Reagent 2 is a mixed solution of dimethyl sulfoxide, disodium hydrogen phosphate, magnesium chloride, sodium azide and sodium nitrite; the mass concentration of dimethyl sulfoxide is 0.1%, the mass concentration of sodium azide is 0.01%, nitrous acid The mass concentration of sodium is 0.01%, the mass concentratio...

Embodiment 2

[0035] Solution preparation and sample detection

[0036] (1) Composition and preparation of the solution

[0037] Reagent 1 is a mixture of diaphorase, nicotinamide adenine dinucleotide and β-hydroxybutyrate dehydrogenase, mixed in a mass ratio of 2:2:1. When preparing, weigh 200mg diaphorase, 200mg nicotinamide adenine dinucleotide and 100mg β-hydroxybutyrate dehydrogenase, mix them evenly and put them in brown plastic, and store them sealed at -20°C for later use.

[0038] Reagent 2 is a mixed solution of dimethyl sulfoxide, disodium hydrogen phosphate, magnesium chloride, sodium azide and sodium nitrite; the mass concentration of dimethyl sulfoxide is 0.1%, the mass concentration of sodium azide is 0.01%, nitrous acid The mass concentration of sodium is 0.01%, the mass concentration of disodium hydrogen phosphate is 1%, the mass concentration of magnesium chloride is 1%, the solvent is sterilized distilled water, and it is stored in a reagent bottle for future use.

[00...

Embodiment 3

[0045] clinical application research

[0046] Select 3 large-scale dairy cow farms in Heilongjiang Province, randomly take the milk samples and blood samples of 500 postpartum lactating cows within 2 months, and process the milk samples according to the milk sample processing method mentioned in the present invention. Kit detection, at the same time prepare ketone powder (an existing detection method for dairy cow ketosis) according to Table 1 and detect milk ketone body content, and compare the two detection methods. Use enzyme-catalyzed kinetic method kit (UK, Randox) to measure the content of β-hydroxybutyric acid in the blood sample. The detection instrument is the Beckman automatic biochemical analyzer in the United States. The content of β-hydroxybutyric acid in the blood sample > 1200 μmol / L is positive Interpretation standard. The test results show that the detection sensitivity of the kit can reach 90%, and the specificity is 92%. The detection result is better than...

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Abstract

The invention relates to a dairy cow ketosis diagnostic kit and a detecting method of the dairy cow ketosis diagnostic kit. The dairy cow ketosis diagnostic kit comprises a reagent 1, a reagent 2, a reagent 3, a detector and a standard colorimetric board, wherein the reagent 1 is a mixture of diaphorase, nicotinamide adenine dinucleotide and beta-hydroxybutyrate dehydrogenase, the reagent 2 is a mixed salutation of dimethyl sulfoxide, disodium hydrogen phosphate, magnesium chloride, sodium azide and sodium nitrite, and the reagent 3 is a thiazolyl blue solution. The detection method of the dairy cow ketosis diagnostic kit comprises the following steps: firstly separating a ketone body from milk, then adding a prepared mixed solution of the reagent 1, the reagent 2 and the reagent 3, and a to-be-detected sample into a feeding hole of the detector, after reacting for 3 minutes, comparing the result with the standard colorimetric board so as to determine a detecting result. The dairy cow ketosis diagnostic kit is convenient to operate, high in specificity and sensitivity, and good in repeatability, thus having a wide market application prospect.

Description

technical field [0001] The invention belongs to the field of veterinary clinical diagnostics, and relates to a diagnostic kit for dairy cow ketosis and a detection method thereof. Background technique [0002] Ketosis is an important nutritional and metabolic disease of dairy cows in the transition period, which has the characteristics of high incidence and clusters. After the pregnant cow gives birth, the feed intake decreases, and the milk production increases rapidly at the same time, the dairy cow is in a state of negative energy balance, causing the occurrence of ketosis in dairy cows. The main features of dairy cow ketosis are elevated levels of acetoacetate, β-hydroxybutyrate and acetone and decreased blood glucose levels. In recent years, due to factors such as the introduction and breeding of high-yielding dairy cow breeds, the development and use of high-energy feed, and the popularization of intensive feeding management, the incidence of ketosis in dairy cows has...

Claims

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Application Information

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IPC IPC(8): G01N21/78
Inventor 张志刚姜惠洁王明河张伟倩姜婧陈雁杨大千于洪祥赵树臣王洪斌
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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