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Multiple rapid detection method for three food borne pathogenic bacteria, and detection primer set and kit thereof

A detection kit and detection primer technology, applied in the field of microbial detection, can solve the problems that have not been seen before, and achieve the effects of strong specificity, high yield of amplification products and high sensitivity

Inactive Publication Date: 2011-10-05
浙江省质量技术监督检测研究院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching domestic and foreign literature, there is no relevant report on the application of LAMP in multiple rapid detection of Salmonella, Shigella and Staphylococcus aureus.

Method used

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  • Multiple rapid detection method for three food borne pathogenic bacteria, and detection primer set and kit thereof
  • Multiple rapid detection method for three food borne pathogenic bacteria, and detection primer set and kit thereof
  • Multiple rapid detection method for three food borne pathogenic bacteria, and detection primer set and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Salmonella, Shigella, Staphylococcus aureus triple LAMP rapid detection of Mg 2+ concentration test

[0034] The concrete detection method of the present embodiment is:

[0035] 1. Extraction of sample DNA

[0036] 1.1. Inoculate the standard strains of Salmonella, Shigella and Staphylococcus aureus in nutrient broth respectively, and incubate at 36°C±1°C for 18 hours.

[0037] 1.2. Use the QIAGEN Bacterial Genomic DNA Extraction Kit to extract the bacterial DNA in the above cultured products respectively.

[0038] 2. LAMP reaction

[0039] 2.1. Artificially synthesize upstream and downstream primers for the detection of Salmonella, Shigella and Staphylococcus aureus respectively, wherein the upstream primer for Salmonella detection has the nucleotide sequence shown in SEQ No.1, and the downstream primer has Nucleotide sequence as shown in SEQ No.2; The upstream primer for Shigella detection has the nucleotide sequence as shown in SEQ No.3, and the downst...

Embodiment 2

[0054] Embodiment 2: Salmonella, Shigella, Staphylococcus aureus triple LAMP rapid detection reaction temperature test

[0055] The concrete detection method of this embodiment is as follows:

[0056] 1. Extraction of sample DNA

[0057] 1.1. Inoculate the standard strains of Salmonella, Shigella and Staphylococcus aureus in nutrient broth respectively, and incubate at 36°C±1°C for 18 hours.

[0058] 1.2. Use the QIAGEN Bacterial Genomic DNA Extraction Kit to extract the bacterial DNA in the above cultured products respectively.

[0059] 2. LAMP reaction

[0060] 2.1. Artificially synthesize upstream and downstream primers for the detection of Salmonella, Shigella and Staphylococcus aureus respectively, wherein the upstream primer for Salmonella detection has the nucleotide sequence shown in SEQ No.1, and the downstream primer has Nucleotide sequence as shown in SEQ No.2; The upstream primer for Shigella detection has the nucleotide sequence as shown in SEQ No.3, and the do...

Embodiment 3

[0075] Example 3: Detection Sensitivity of Triple LAMP Rapid Detection to Salmonella

[0076] The concrete detection method of the present embodiment is:

[0077] 1. Extraction of sample DNA

[0078] 1.1. Inoculate the standard strain of Salmonella in nutrient broth and incubate at 36°C±1°C for 18 hours.

[0079] 1.2. Use the QIAGEN Bacterial Genomic DNA Extraction Kit to extract the bacterial DNA in the above cultured products respectively.

[0080] 2. LAMP reaction

[0081] 2.1. Artificially synthesize upstream and downstream primers for the detection of Salmonella, Shigella and Staphylococcus aureus respectively, wherein the upstream primer for Salmonella detection has the nucleotide sequence shown in SEQ No.1, and the downstream primer has Nucleotide sequence as shown in SEQ No.2; The upstream primer for Shigella detection has the nucleotide sequence as shown in SEQ No.3, and the downstream primer has the nucleoside as shown in SEQ No.4 acid sequence; the upstream prim...

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Abstract

The invention discloses a multiple rapid detection method for three food borne pathogenic bacteria, and detection primer sets and a detection kit thereof. An upstream primer and a downstream primer of a rapid detection primer set for Salmonella have sequences as represented by SEQ No.1 and SEQ No.2, respectively; an upstream primer and a downstream primer of a rapid detection primer set for Shigella have sequences as represented by SEQ No.3 and SEQ No.4, respectively; an upstream primer and a downstream primer of a rapid detection primer set for o Staphylococcus aureus have sequences as represented by SEQ No.5 and SEQ No.6, respectively. Bacterial genome DNA extracted from a sample to be tested is subjected to a LAMP reaction with the primer sets in a same reaction system, and the resultant of the reaction is identified to determine whether the sample contains Salmonella, Shigella or Staphylococcus aureus. Every primer set provided in the invention has strong specificity and can accurately detect genome DNAs of Salmonella, Shigella and Staphylococcus aureus in a same reaction system.

Description

technical field [0001] The invention belongs to the field of microbial detection, and relates to a triple rapid detection method for Salmonella, Shigella and Staphylococcus aureus, a detection primer set and a detection kit. Background technique [0002] salmonella( Salmonella spp. ) is an important pathogen in the field of public health, belonging to Gram-negative Enterobacteriaceae. According to statistics, in the bacterial food poisoning that occurs in countries all over the world, the food poisoning caused by Salmonella often ranks first, and the bacterial food poisoning that occurs in the inland areas of our country also takes Salmonella as the primary cause. In addition to causing food poisoning, bacteria in this genus can also cause diseases such as gastroenteritis, typhoid fever, and paratyphoid fever. Shigella( Shigella spp ) is a Gram-negative Enterobacteriaceae, the most common pathogen of human bacillary dysentery. In recent years, Shigella type Ⅰ bacillary ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCY02A50/30
Inventor 姜侃张东雷金燕飞黄建锋汪新李洪波
Owner 浙江省质量技术监督检测研究院
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