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PCR primer pair for identifying or assisting in identifying tissue and/or organ of cat and application thereof

A primer pair and cat identification technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of antibody specificity, inability to detect heat-processed meat, and mixed meat detection cross-reaction issues

Inactive Publication Date: 2012-03-21
CHINA ANIMAL DISEASE CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of identification technology is greatly affected by the structure of the protein (the part that plays a major role is the antigenic determinant), and the problems that often occur in practical applications include: 1. The thermal processing process will change the structure of the protein (the antigenic determinant). Detect heat-processed meat; 2. Cross-reaction may occur in the detection of mixed meat, because the structure of proteins of different species may have a certain degree of similarity, and the sensitivity of the test depends on the specificity of the prepared antibody
The DNA molecular hybridization method is complicated to operate, and the cost is high, so it is gradually replaced by the PCR method

Method used

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  • PCR primer pair for identifying or assisting in identifying tissue and/or organ of cat and application thereof
  • PCR primer pair for identifying or assisting in identifying tissue and/or organ of cat and application thereof
  • PCR primer pair for identifying or assisting in identifying tissue and/or organ of cat and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, detect animal pure muscle sample

[0056] 1. Preparation of PCR primer pairs for identification or auxiliary identification of feline tissues and / or organs

[0057] Taking cat meat mitochondrial 16S ribosomal RNA gene as the target gene, a primer pair was designed to specifically amplify the gene. The sequence of the upstream primer is 5'-AGCGCAACAATCAAAGCATCT-3' (sequence 1 in the sequence listing), and the sequence of the downstream primer is 5'-CGCGGCCGTTTAACTAGC-3' (sequence 2 in the sequence listing). The primer pair is the PCR primer pair for identifying or assisting in identifying cat tissues and / or organs. The primer pair amplifies the target sequence of domestic cat as shown in sequence 11 in the sequence listing. The 130th-135th position of sequence 11 in the sequence listing is the recognition sequence of Hind III, and the PCR product will become two fragments after enzyme digestion, the lengths are 132bp and 235bp respectively.

[0058] The ...

Embodiment 2

[0090] Embodiment 2 detects doped muscle sample

[0091] 1. Preparation of PCR primer pairs for identification or auxiliary identification of animal tissues and / or organs

[0092] The design and preparation method are the same as in Example 1.

[0093] 2. Detection of animal muscle samples

[0094] 1. Preparation of test samples

[0095] The samples in this experiment were min pork, yellow beef, goat and local chicken adulterated with domestic cat meat:

[0096] 1) Pork mixed with cat meat:

[0097] 0.1 g of cat meat is mixed in 99.9 g of pork to make 0.1% pork mixed with cat meat.

[0098] 2) Beef mixed with cat meat:

[0099] Add 0.1g of cat meat in 99.9g of beef to make 0.1% beef mixed with cat meat.

[0100] 3) Mutton mixed with cat meat:

[0101] Add 0.1g of cat meat to 99.9g of mutton to make 0.1% mutton mixed with cat meat.

[0102] 4) Chicken mixed with cat meat:

[0103] Add 0.1g of cat meat to 99.9g of chicken to make 0.1% chicken mixed with cat meat.

[01...

Embodiment 3

[0116] Embodiment 3, Utilize the PCR primer pair of identification or auxiliary identification cat tissue and / or organ to detect different cat meat samples

[0117] 1. Preparation of PCR primer pairs for identification or auxiliary identification of feline tissues and / or organs

[0118] The design and preparation method are the same as step 1 of Example 1.

[0119] 2. Detection of various cat meat samples

[0120] 1. Test samples

[0121] The test samples were the muscles of long-haired cats, short-haired cats and short-haired tiger cats.

[0122] 2. Using the primer pair in step 1 to PCR amplify the 16S ribosomal RNA gene fragment on the mitochondrial genome

[0123] 1) Extraction of sample genome

[0124] With embodiment 1.

[0125] 2) PCR reaction

[0126] With embodiment 1.

[0127] The products obtained by the PCR reaction were respectively subjected to agarose gel electrophoresis, and the concentration of the agarose gel electrophoresis was 2%. Electrophoresis de...

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Abstract

The invention discloses a PCR (Polymerase Chain Reaction) primer pair for identifying or assisting in identifying a tissue and / or an organ of a cat and application thereof. In five PCR primer pairs for identifying the tissue and / or the organ of the cat, a cattle, a sheep, a pig or a chicken, the primer pair provided by the invention comprises at least one primer pair of the five PRC primer pairs for identifying the tissue and / or the organ of the cat; the five PCR primer pairs for identifying the tissue and / or the organ of the cat, the cattle, the sheep, the pig or the chicken sequentially andrespectively consist of two DNA single strands shown by a sequence 1 and a sequence 2, two DNA single strands shown by a sequence 3 and a sequence 4, two DNA single strands shown by a sequence 5 and a sequence 6, two DNA single strands shown by a sequence 7 and a sequence 8, and two DNA single strands shown by a sequence 9 and a sequence 10. The five primer pairs of the invention all choose the annealing temperature of 62 DEG C, so that all the identifications can be completed by the PCR at a time at the same temperature, in addition, the specificity is strong, and the time spent on the detection process is short.

Description

technical field [0001] The present invention relates to a pair of PCR primers for identifying or assisting in the identification of animal tissues and / or organs and applications thereof, in particular to identifying or assisting in the identification of at least one animal tissue and / or organ in the five species of cats, cattle, sheep, pigs, and chickens or PCR primer pairs for organs. Background technique [0002] At present, in the production and sales of domestic raw meat and meat products, in order to make huge profits, some unscrupulous traders use adulteration and other means to deceive consumers from time to time, such as using cat meat as pork, beef and mutton. Therefore, a set of methods for quickly and accurately identifying meat types and being able to adapt to the identification of mixed meat products will undoubtedly provide strong technical support for law enforcement officers. [0003] The traditional identification methods of meat products are mainly based o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10
Inventor 于雷姜艳彬王海李通尹艳侯东军杨红菊李颖
Owner CHINA ANIMAL DISEASE CONTROL CENT
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