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Kit for detecting cow milk content in goat milk product and application of kit

A kit and goat milk technology, applied in the field of food safety, can solve the problems of low cell content and inability to judge negative results, etc., and achieve the effects of strong specificity, good reproducibility, and low detection limit

Inactive Publication Date: 2015-06-17
黑龙江省乳品工业技术开发中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The content of cells in goat milk products is relatively low. According to literature reports, during the processing of dairy products, DNA fragments will be broken, from 1100bp to about 600bp. Therefore, when the content of target animal-derived components in the test object is very low, It is often impossible to determine whether the negative result is caused by the loss of DNA due to the process

Method used

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  • Kit for detecting cow milk content in goat milk product and application of kit
  • Kit for detecting cow milk content in goat milk product and application of kit
  • Kit for detecting cow milk content in goat milk product and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1: experimental method

[0048] 1) Preparation of Standards

[0049] Collect goat milk and cow milk control substances, freeze-dry, and prepare goat milk containing 5 kinds of cow milk concentrations such as 100%, 50%, 25%, 10% and 1% (W / W) as standard substances for control, Total DNA was extracted, aliquoted and stored at -20°C for standard curve.

[0050] 2) Samples to be inspected

[0051] Commercially available goat milk powder was used as the test sample, and DNA was extracted with reference to Blood & Tissue Handbook.

[0052] 3) Primer and probe design and synthesis

[0053] The specific primers and probes of the internal control gene were designed for the homology sequence of 12SrRNA of cattle and sheep, and the primers and probes of specific genes of cattle were designed for the partial nucleotide sequence (SEQ ID NO.7) of 12SrRNA of cattle. Needle.

[0054] 12SrRNA is located on the mitochondria, which have the characteristics of high temper...

Embodiment 2

[0077] Embodiment 2: the analysis of inspection result

[0078] 1) Determination of standard curve and detection limit

[0079] Collect goat milk and cow milk control substances, freeze-dry, prepare goat milk containing 5 cow milk concentrations of 100%, 50%, 25%, 10%, 1% (W / W) and other control substances, and extract total DNA , taking the logarithmic values ​​of the five standard concentrations as the horizontal axis, and the △Ct obtained by subtracting the average Ct value of the internal control gene Ct value from the average Ct value of the bovine-specific gene of each concentration standard product as the vertical axis, each concentration standard The product needs to be tested three times, and the difference between the Ct values ​​of the parallel samples should be <0.5. Outliers should be discarded to ensure that there are representative data for each concentration before being included in the analysis and calculation.

[0080] The obtained formula model is: y=ax 2+...

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Abstract

The invention discloses a kit for detecting cow milk content in a goat milk product and application of the kit, and belongs to the technical field of food safety. The kit comprises a specific primer and a probe for internal comparison genes as well as a primer and a probe for specific genes of a cow, wherein the specific primer for the internal comparison genes is designed and obtained after comparing the homologous sequences of 12SrRNA of the cow and a goat; the primer for the specific genes of the cow is designed for the partial nucleotide sequence of the 12SrRNA of the cow. The invention further provides a method for detecting the cow milk content in the goat milk product. The method comprises the following steps: extracting the DNA of a sample; utilizing the primer and probe for internal comparison as well as the primer and probe for the specific genes to carry out an FQ-PCR amplification reaction; quantitatively detecting the cow milk content in the goat milk product. The method is accurate, reliable, high in specificity, low in detection limit, high in reproducibility and suitable for rapid and quantitative detecting of the cow milk content in the goat milk product.

Description

technical field [0001] The invention relates to a kit for detecting milk content in goat milk products and its application, which belongs to the technical field of food safety. Background technique [0002] In some developed countries, goat milk products are high-end products in the dairy market, mainly consumed by high-income families. Although the price of goat milk and its products is relatively high, its market share has remained high. Australia and New Zealand are key goat milk export countries and regions. Goat milk consumption in Southeast Asia is mainly in Singapore, Malaysia, Hong Kong, Taiwan and other places. In Taiwan Province of China, the consumption of goat milk is relatively large, and the demand is also relatively large. Taiwanese generally recognize the advantages of goat milk being close to breast milk, especially Goat milk formula for infants and young children in Taiwan has been accepted by the majority of parents. According to the survey, among the wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q1/686C12Q2600/158C12Q2600/166C12Q2545/101
Inventor 卢雁鄂志强沙淼刘春波
Owner 黑龙江省乳品工业技术开发中心
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