Detection method for infant salmonella PCR and nucleic acid and primer pair thereinto

A Salmonella infantis and detection method technology, applied in the field of Salmonella infantis PCR detection method and nucleic acid and primer pairs therein, can solve the problems such as the nucleic acid and primers of the PCR detection method for Salmonella infantis serotypes which have not been invented, increase detection complexity, complex coding genes, etc. , to achieve the effect of practicability, short detection time and specific detection results

Active Publication Date: 2010-06-23
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the polymorphism of Salmonella antigens and the complexity of the coding genes, it is difficult to find suitable target genes and detection primer pairs, and for some serotype strains with similar antigenic structures to Salmonella infantis, it is necessary to combine other related genes to form a multiplex PCR can only be determined, which increases the complexity of detection
[0004] Found through literature retrieval to prior art, as yet invention and the report relevant to the PCR detection method of infant Salmonella serotype of the present invention, nucleic acid and primer

Method used

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  • Detection method for infant salmonella PCR and nucleic acid and primer pair thereinto
  • Detection method for infant salmonella PCR and nucleic acid and primer pair thereinto
  • Detection method for infant salmonella PCR and nucleic acid and primer pair thereinto

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Establishment of PCR detection method for infant Salmonella serotype

[0023] Step 1, design amplification primers according to the conserved sequence in the genomic DNA sequence of Salmonella infantis

[0024] Find the specific restriction endonuclease gene from the Salmonella infantis genome DNA sequence, and use it as the detection target gene of Salmonella infantis, the gene sequence is shown in SEQ ID NO: 1;

[0025] Input the DNA sequence of the restriction endonuclease gene into the primer design software Primer Premier 5.0 to design primers, set the GC% range to 40-60%, and the product size range to 150-300bp, and select primers from the alternative primer pairs , the primer sequences are as follows (primers were synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.):

[0026] SIN5-L: 5'-AGCCAACGCCACCTACTACT-3' (SEQ ID NO: 2);

[0027] SIN5-R: 5'-TGAACACCATATCCATCCACAT-3' (SEQ ID NO: 3);

[0028] Step 2, DNA template preparation

[0029]...

Embodiment 2

[0042] Detection of suspected strains of Salmonella

[0043] Using the PCR detection method for infant Salmonella serotypes in Example 1, 53 suspected strains of Salmonella isolated from food samples were detected. The food samples were collected in suburban supermarkets and bazaars in Shanghai. For sample processing and isolation of suspected strains, refer to the national standard GB / T 4789.4-2008.

[0044] From it, 2 suspected strains were detected to be positive results, and these 2 suspected strains were identified as O7:Hr:5 by the Salmonella diagnostic serum (Lanzhou Institute of Biological Products), and it was determined that it was Salmonella infantis (see the product manual for the serum identification steps and National standard GB / T 4789.4-2008), other suspected strains were identified as not Salmonella infantis. This example proves that the PCR detection method for Salmonella infantis has very high reliability.

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Abstract

The invention relates to a detection method for infant salmonella PCR and nucleic acid and primer pair therein, belonging to the technical field of food safety detection; the detection method comprises the following steps: designing an amplimer according to the conserved sequence SEQ ID NO:1 in the genome DNA sequence of the infant salmonella; extracting sample DNA, amplifying by PCR method; detecting the amplified product through gel electrophoresis, and judging whether the sample contains the infant salmonella or not; and the judgment is particular as follows: if corresponding single amplification belt does not appear, the sample does not contain the infant salmonella. The invention also relates to a nucleic acid, the base sequence of which is showed by SEQ ID NO:1; the invention also relates to a pair of primer, particularly the base sequence of which is showed by the nucleic acid SEQ ID NO: 2, and SEQ ID NO: 3. When detecting the infant salmonella by adopting the invention, the detection time is short, the cost is low, the detection result is distinctive and the result is judged easily.

Description

technical field [0001] The invention relates to a detection method in the technical field of food safety inspection and a nucleic acid and a primer pair therein, a PCR detection method for infant Salmonella and the nucleic acid and a primer pair therein. Background technique [0002] Salmonella infantis (Salmonella infantis) is one of the main Salmonella serotypes that cause infection in newborns and infants. It was first discovered by Wheele and Borman in 1943. The main sources of infection for Salmonella infants are animals (mainly poultry), food, and people carrying bacteria. They can infect healthy humans through food, environment, etc. through the oral cavity, reproduce in their intestines, invade the intestinal wall mucosa, and cause enteritis, Dysentery, sepsis and other diseases. Salmonella infection in infants has no strict seasonality and can occur in all seasons. It has strong resistance in polluted environments and can survive for several months. Therefore, it w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/31C12N15/11C12R1/42
Inventor 史贤明刘斌施春雷何晓华
Owner SHANGHAI JIAO TONG UNIV
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