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PCR (Polymerase Chain Reaction) detection method of salmonella enteritidis, nucleic acid and primer pair

A Salmonella Enteritidis and detection method technology, which is applied in the field of food safety detection, can solve problems such as the PCR detection method of Salmonella Enteritidis that has not yet been invented, and achieve the effects of simple result determination, reliable detection results, and reduced detection costs

Active Publication Date: 2010-10-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Through literature search to the prior art, it is found that there is no report related to the PCR detection method, nucleic acid and primer pair of Salmonella enteritidis of the present invention.

Method used

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  • PCR (Polymerase Chain Reaction) detection method of salmonella enteritidis, nucleic acid and primer pair
  • PCR (Polymerase Chain Reaction) detection method of salmonella enteritidis, nucleic acid and primer pair
  • PCR (Polymerase Chain Reaction) detection method of salmonella enteritidis, nucleic acid and primer pair

Examples

Experimental program
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Embodiment 1

[0020] Salmonella enteritidis PCR detection method

[0021] Step 1, designing primers according to the conserved sequence in the genomic DNA sequence of Salmonella enteritidis

[0022] Through comparative genome and bioinformatics analysis, a specific DNA sequence was found in the genome DNA sequence of Salmonella Enteritidis, and it was used as a detection target of Salmonella Enteritidis. The base sequence of this nucleic acid is shown in SEQ ID NO: 1;

[0023] This DNA nucleic acid sequence is imported into the primer design software Primer Premier 5.0 to design primers, the GC% range is set to 40-60%, the product size range is 150-1000bp, and the primers are selected from the alternative primer pairs. The primer sequences are as follows (primers Synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.):

[0024] SEN2-L: 5'-GCCACTGTCTGATGCTCTTG-3', (SEQ ID NO: 2)

[0025] SEN2-R: 5'-GAAAGGCTCCGTGGTTAGT-3'; (SEQ ID NO: 3)

[0026] Step 2, the system and ...

Embodiment 2

[0038] Detection of suspected strains of Salmonella enteritidis

[0039] Using the PCR detection method for Salmonella enteritidis established in Example 1, 63 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. Detected 4 suspected bacterial strains were positive results, these 4 suspected bacterial strains were identified as 09:Hg, m:7 by Salmonella diagnostic serum (Lanzhou Institute of Biological Products), and determined to be Salmonella enteritidis (serum identification steps please refer to the product Instructions and national standard GB / T 4789.4-2008), other suspected strains were identified as not Salmonella Enteritidis. This example proves that the PCR detection method for Salmonella enteritidis has very high reliability.

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Abstract

The invention discloses a PCR (Polymerase Chain Reaction) detection method of salmonella enteritidis, nucleic acid and a primer pair in the technical field of food safety detection. The PCR (Polymerase Chain Reaction) detection method comprises the following steps of: designing a specific amplimer pair according to base sequence in genomic DNA of the salmonella enteritidis, such as the sequence shown as in SEQ ID NO:1; extracting a sample DNA and amplifying by a PCR method; and detecting an amplified product by gel electrophoresis and judging whether the sample contains the salmonella enteritidis or not, wherein the judgments are specifically as follows: detecting whether the amplified product has single amplified strip or not in a 656bp position, if so, judging that the sample contains the salmonella enteritidis, and if not, judging that the sample contains no salmonella enteritidis. The invention also relates to the nucleic acid, the base sequence of which is shown as the SWQ IDNO:1, and the primer pair, wherein the base sequence of a forward primer is shown as SEQ ID NO:2, and the base sequence of a reverse primer is shown as SEQID NO:3. By adopting the detection method to detect the salmonella enteritidis, the detection time is short, the cost is low, the practicability is better, the detection result is peculiar and the judging result is simple.

Description

technical field [0001] The invention relates to a PCR detection method, a nucleic acid and a primer pair in the technical field of food safety detection, in particular to a PCR detection method, a nucleic acid and a primer pair for Salmonella enteritidis. Background technique [0002] Salmonella Enteritid (Salmonella Enteritid) is one of the most common types of Salmonella infection. It has no host specificity and is very aggressive. It can cause gastroenteritis in livestock and poultry, enteritis and food poisoning in humans. Salmonella enteritidis is highly resistant to the external environment and can survive for several months in water, milk and meat products. Salmonella Enteritidis is widely distributed in nature and is stored in the intestines of many poultry and wild animals, among which poultry eggs and poultry meat are the main sources of infection for humans. Clinically, the main symptoms of Salmonella Enteritidis infection are fever, nausea, vomiting and diarrhea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/10C12N15/11
CPCY02A50/30
Inventor 史贤明刘斌施春雷何晓华陈婧
Owner SHANGHAI JIAO TONG UNIV
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