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Quick experimental animal resource quality monitoring method

A technology of experimental animals and resources, applied in the field of biology, can solve the problems of low specificity and time-consuming

Inactive Publication Date: 2017-01-04
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods are time-consuming, or have low specificity, and require the inspectors to have accurate judgments on the colony morphology and physical and chemical reaction properties of pathogenic microorganisms before they can draw correct conclusions, which requires the inspectors to have solid microbiological knowledge Theoretical basis and rich experience in pathogenic microorganism detection

Method used

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  • Quick experimental animal resource quality monitoring method
  • Quick experimental animal resource quality monitoring method
  • Quick experimental animal resource quality monitoring method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1, formulation of experimental technical route and utilization of positive standard bacterial strains to screen primers

[0060] 1. Establishment of PCR method for positive control

[0061] The target microorganisms to be detected in this experiment are: Staphylococcus aureus, beta-hemolytic streptococcus, and Klebsiella pneumoniae.

[0062] Purchase the above three standard strains from the China Medical Microbial Culture Collection Center (CMCC) as positive controls, resuscitate these three standard strains, extract the DNA of the revived standard strains, and design corresponding to each standard strain according to the gene sequence The specific primers were selected, the corresponding primers were screened and the PCR system was optimized, and the PCR products were sequenced and compared to verify the accuracy of the positive standard strains, so as to establish the PCR method. Process such as figure 1 .

[0063] 2. Primer screening

[0064] After a ...

Embodiment 2

[0084] Example 2, positive standard strain PCR result product sequencing result BLAST

[0085] Electrophoresis and sequencing were performed on the obtained positive standard strain PCR products. The obtained sequencing results were subjected to BLAST. The parameters are as follows:

[0086] Score is the alignment score, the higher the similarity between two sequences;

[0087] The smaller the E Value, the more credible it is;

[0088] Identities is consistency, which means that the two sequences maintain the same ratio.

[0089] 1. Staphylococcus aureus primer 1

[0090] Using Staphylococcus aureus primer 1, the target band 122bp was obtained. Blast results such as image 3 shown.

[0091] 2. Staphylococcus aureus primer 2

[0092] Using Staphylococcus aureus primer 2, the target band 132bp was obtained. Blast results such as Figure 4 shown.

[0093] 3. Klebsiella pneumoniae primer 1

[0094] Klebsiella pneumoniae primer 1 was used to obtain the target band 368bp...

Embodiment 3

[0102] Example 3, establishment of a rapid detection method for frozen resources

[0103] The frozen resources (embryo and sperm, etc.), the solution required in the freezing and recovery process, and the sterilized water containing liquid nitrogen for preserving frozen storage were used to extract DNA with a micro DNA extraction kit (purchased from QIAGEN).

[0104] PCR primers and corresponding PCR conditions and PCR reaction system for Staphylococcus aureus, Klebsiella pneumoniae, beta-hemolytic streptococcus provided in the foregoing embodiment 1 are carried out to the above-mentioned sample DNA for PCR reaction, if a positive strip is obtained If there is a band, it can prove that the corresponding strain exists in the frozen resources, and at the same time, the PCR product can be recovered for further confirmation by DNA sequencing.

[0105] Operation process such as figure 2 shown.

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Abstract

The invention relates to a quick experimental animal resource quality monitoring method and discloses a method for specifically identifying the target microbial infection situation of an experimental animal resource. A PCR primer having very good specificity is obtained through screening, and the quick experimental animal resource quality monitoring method has detection sensitivity and accuracy.

Description

technical field [0001] The invention belongs to the field of biology, and more specifically, the invention relates to a method for quickly monitoring the quality of experimental animal resources. Background technique [0002] The application of cryogenic technology for experimental animal resources (embryos, sperm, etc.) has many advantages. In medical biological research, various strains of experimental animals are needed, and maintaining animal strains in isolators requires a lot of manpower and material resources, and is expensive. If the temporarily unused animal resources are frozen, they can be preserved for a long time, which is safe and saves manpower and material resources. The use of animal resource transportation instead of live animal transportation is convenient and can reduce the spread of diseases, and facilitate the international exchange of experimental animal strains. In addition, valuable genetic material can be preserved, a gene pool can be established,...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/689C12Q2600/16
Inventor 强苏静刘丽均顾剑洁赵立虎徐平
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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