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Five-fold PCR detection method and detection kit for avian Escherichia coli, Salmonella gallinarum typhi, Salmonella pullorum, etc.

A technology for Salmonella typhi and Salmonella pullorum is applied in the biological field to achieve the effects of sensitive detection, high sensitivity and strong specificity

Active Publication Date: 2021-03-16
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, a variety of detection methods have been established for avian Escherichia coli and different serotypes of Salmonella, however, there are no rapid detection and identification methods for avian Escherichia coli, Salmonella gallinarum typhi, Salmonella pullorum, Salmonella enteritidis, and Salmonella typhimurium Diagnostic PCR method

Method used

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  • Five-fold PCR detection method and detection kit for avian Escherichia coli, Salmonella gallinarum typhi, Salmonella pullorum, etc.
  • Five-fold PCR detection method and detection kit for avian Escherichia coli, Salmonella gallinarum typhi, Salmonella pullorum, etc.
  • Five-fold PCR detection method and detection kit for avian Escherichia coli, Salmonella gallinarum typhi, Salmonella pullorum, etc.

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 5

[0038] Example 1 Five-Demand PCR Detection Method

[0039] This example is used for multiple PCR to detect poultry, chicken injury, chicken, chicken, sofan, salamobacteria, mouse injury, chicken, chicken, chicken, chicken, chicken, chicken, Dock Salmonella, enteritis, enteritis Salmonica and murine are taking the soforma for testing, including the following steps:

[0040] Step 1, prepare a PCR template based on the bacterial to be detected

[0041] The bacterial strain to be inspected is inoculated to the LB liquid medium, which is cultured to OD at a temperature of 37 ° C. 600 = 1.0, that is, the bacterial liquid that can be used as a PCR template can be obtained.

[0042] Step 2, prepare a specific primer group

[0043] Preparation for multiple PCR detection of poultry coli, chicken injury, sofavy, chicken white snorcium, salamobacteria, mouse injury, sofama: The concentration of the concentration of 10 pm is 10 pm, dilution of this concentration The liquid is placed at -20 ° C...

Embodiment 2 5

[0071] Example 2 Five-five PCR detection kit for detection

[0072] In this embodiment, the above-mentioned PCR reaction system is established by the present embodiment directly employing a five-member PCR detection kit including the primer group mentioned in Example 1.

[0073] The composition of the five-membered PCR detection kit included 2 × PCR premix and primer groups, wherein the volume of 2 × PCrPremix was 12.5 μL, and the primer in the primer group was 0.5 μl of ECPHOA-F and ECPHOA-R. The volume of SGGLGC-F and SGGLGC-R is 0.2 μL, the volume of the primer to SGPSPEC-F and SGPSPSPEC-R is 0.4 μl of SSDFI-F and SESDFI-R, and the primer is StSTM4495. The volume of -f and STSTM4495-R is 0.5 μL.

[0074] Evaluation Example 1 Specific Evaluation

[0075] In this example, the five-member PCR detection kit of Example 2 specifically detected the primer group in Example 1 and Example 2:

[0076] Choose the duck hyperplate bacillus, golden staphylococcus, copper green psessa pA14, po...

Embodiment 1

[0105] Embodiment 1 and Effect of Example 1

[0106] Through the evaluation examples 1 to the evaluation example 3, it can be seen that due to the amplification of the positive strains that have been preserved in the laboratory, they can be amplified to the specific strip, while other bacterial strains, etc. To the strip, a positive sample of the primer group is indicated, and a positive sample of different dilutions is detected using the primer group, and the minimum detection sensitivity can reach 1000cFu bacterial liquid and 500 pg bacterial genome DNA, indicating the primer set. High sensitivity, at the same time, the primer group has good repetition, indicating that its stability is strong. Therefore, Examples 1 and Example 2 included a five-member PCR detection method using the primer set or a five-member PCR detection kit containing the above-mentioned primer group, the poultry, chicken injury, chicken, chicken, Dock, Salmonella, enteritis Shamen The detection of the bacter...

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Abstract

The invention provides a multiple PCR primer group for avian escherichia coli, chicken typhoid salmonella, salmonella pullorum, salmonella enteritidis and bacillus breslaviensis, a quintuple PCR detection kit comprising the primer group and a quintuple PCR detection method. The primer group comprises primer pairs ECphoA-F and ECphoA-R for specific amplification of avian escherichia coli phoA genes, primer pairs SGg1gC-F and SGg1gC-R for specific amplification of chicken typhoid salmonella glgC genes, primer pairs SGPspeC-F and SGPspeC-R for specific amplification of chicken typhoid salmonella and salmonella pullorum genes, primer pairs SEsdf I-F and SEsdf I-R for specific amplification of salmonella enteritidis sdf-1 genes and primer pairs STstm4495-F and STstm4495-R for specific amplification of bacillus breslaviensis stm4495 genes.

Description

Technical field [0001] The present invention belongs to the field of biotechnology, and more particularly to multiple PCR primer groups for detecting poultry E. coli, chickens, chicken, sofama, salamobacteria, mouse injury, including the primer group five PCR The kit and five PCR detection methods utilized using the primer set. Background technique [0002] With the rapid development of scale, the occurrence and epidemic of the diseased, it also has increased accordingly, leading to a decline in economic benefits of the poultry industry. Important bacterial infectious diseases such as poultry, chicken, chicken, chicken, chicken, etc., are seriously harmful to poultry. [0003] At present, China mainly relying on antibiotic prevention and treatment of bacterial disease, but due to long-term, unreasonable use of antibiotics: On the one hand, a wide range of drug resistance is produced, and the drug residue directly affects the safety of animal products. Therefore, it is important t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/10C12N15/11C12R1/19C12R1/42
CPCC12Q1/686C12Q1/689C12Q2600/16C12Q2531/113
Inventor 于圣青王少辉梁华吴晓君丁铲
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI