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Integrated detection method for salmonella and serotype thereof

A technology for serotype detection and Salmonella, which is applied in microorganism-based methods, biochemical equipment and methods, and microbial determination/inspection. and material resources, etc., to achieve the effect of convenient and accurate result judgment, fast detection speed, and easy operation.

Pending Publication Date: 2016-12-21
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of the above-mentioned bacteriological culture detection method are: (1) Bacteria enrichment requires a long culture process, and it takes at least 4 days to obtain a positive result, 7 days to confirm a negative result, and a longer time for repeated confirmation and verification. Therefore, the detection cycle is long and the workload is heavy
(2) The cost is high, consuming manpower and material resources
(3) When determining the serotype of Salmonella, the serotype interpretation process is cumbersome and requires the subjective judgment of the operator, resulting in large errors and poor repeatability
So far there have been reports that PCR amplification technology has been used for qualitative detection of Salmonella, but the sensitivity and specificity of the primer pairs used for amplification in practical applications are still unsatisfactory, and there is no use of existing primers so far. A report on the rapid determination of Salmonella serotypes that can be directly used for the detection of gene sequences after PCR amplification

Method used

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  • Integrated detection method for salmonella and serotype thereof
  • Integrated detection method for salmonella and serotype thereof
  • Integrated detection method for salmonella and serotype thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1: the method for one-step identification of Salmonella and its serotype, comprising the following steps:

[0067] Sampling of samples is preferred: 3 meat food samples were collected in Auchan Supermarket in Minhang District, Shanghai, enriched and cultured, and 9 strains to be tested were obtained.

[0068] Step 1: DNA was extracted from the 9 strains to be tested obtained above using the CTAB method, Salmonella typhi was used as a positive control, and deionized purified water was used as a negative control.

[0069] Step 2: using the DNA extracted in step 1 as a template, and performing PCR amplification under the guidance of primer pair S9;

[0070] The reaction system of PCR amplification is: (NH4) 2 SO 4 Buffer (10x): 2.5 μL; MgCl 2 Solution (50mmol / L): 1.5μL; dNTP (2.5mmol / L): 1.5μL; upstream and downstream primers (each 10μmol / L): 0.5μL+0.5μL; Taq DNA polymerase (1U / μL): 1.0μL; Template: 2.5 μL; Sterile deionized water: 15 μL; Total volume: 25 μL...

Embodiment 2

[0084] Embodiment 2: the method for one-step identification of Salmonella and its serotype, comprising the following steps:

[0085] Sampling of samples is preferred: 2 samples of milk and 2 samples of egg food were collected at Auchan Supermarket in Minhang District, Shanghai, enriched and cultured, and 9 strains to be tested were obtained.

[0086] Step 1: DNA was extracted from the 9 strains to be tested obtained above using the CTAB method, Salmonella typhi was used as a positive control, and deionized purified water was used as a negative control.

[0087] Step 2: using the DNA extracted in step 1 as a template, and performing PCR amplification under the guidance of primer pair S69;

[0088] The reaction system of PCR amplification is: (NH 4 ) 2 SO 4 Buffer (10x): 2.5 μL; MgCl 2 Solution (50mmol / L): 1.5μL; dNTP (2.5mmol / L): 1.5μL; upstream and downstream primers (each 10μmol / L): 0.5μL+0.5μL; Taq DNA polymerase (1U / μL): 1.0μL; Template: 2.5 μL; Sterile deionized water...

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Abstract

The invention discloses a detection primer for salmonella and a serotype thereof, a method for detecting the salmonella and serotype by means of the primer and a kit. Through the primer and the detection method, typing detection can be quickly performed on the salmonella and salmonella serotype in an integrated manner. The integrated detection method for the salmonella and serotype has the advantages of quick and effective detection method, low cost, simple operation, high sensitivity and the like, thus providing a new technical platform for integrated detection of the salmonella and serotype thereof. The integrated detection method disclosed by the invention can be used by stock farming production units, primary medical and health units and disease prevention and control centers for screening and detecting the salmonella and serotype thereof, has broad market prospects and great economic and social benefits, and is suitable for wide-range popularization.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a primer pair for detecting and identifying Salmonella and its serotype and a method for detecting and identifying Salmonella and its serotype. Background technique [0002] Salmonella is one of the most important foodborne pathogens causing food poisoning and can be transmitted between humans and animals. Food products contain a variety of rich nutrients, which are very suitable for the growth and reproduction of Salmonella. Once people ingest animal products containing a large amount of Salmonella, they will cause bacterial infections, and then food poisoning will occur under the action of toxins. Salmonella not only endangers human health, but also brings huge economic losses to animal husbandry. Diseases caused by Salmonella are mainly divided into two categories: one is typhoid and paratyphoid, and the other is acute gastroenteritis. People infecte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/10C12N15/11C12R1/42
CPCC12Q1/689
Inventor 史贤明石秀清周秀娟施春雷崔妍
Owner SHANGHAI JIAO TONG UNIV