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Shigella sonnei special nucleotide PCR detection kit

A nucleotide and shigella technology, which is used in the determination/inspection of microorganisms, DNA/RNA fragments, biochemical equipment and methods, etc., can solve the problems of difficult identification, no rapid identification method, etc., and achieve high detection accuracy. , the market application prospects are wide, the effect of eliminating the extraction step

Inactive Publication Date: 2018-08-10
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Among the several known serotypes of P. shigella-like, O17 is difficult to identify from Shigella sonnei, and there is no rapid identification method

Method used

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  • Shigella sonnei special nucleotide PCR detection kit
  • Shigella sonnei special nucleotide PCR detection kit
  • Shigella sonnei special nucleotide PCR detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0036] 1: Genome extraction

[0037] (1) In the ultra-clean workbench, take out the strain from the strain preservation room, thaw it on ice, inoculate it into LB medium at a ratio of 1:1000, and cultivate overnight at 37°C on a shaker at 180rpm.

[0038] (2) Take 1 mL of the bacterial suspension into a 1.5 mL centrifuge tube, centrifuge at 8000 rpm for 5 minutes, and discard the supernatant.

[0039] (3) Add 250 μL of 50 mM Tris-HCl buffer (pH 8.0) to resuspend, centrifuge at 12,000 rpm for 5 minutes, discard the supernatant, and repeat once.

[0040] (4) Add 250 μL of 50 mM Tris-HCl buffer (pH 8.0) to resuspend, add 10 μL of 0.45M EDTA (pH 8.0), fully suspend, and incubate at 37°C for 20 minutes.

[0041] (5) Add 10 μL of 20 mg / mL lysozyme and incubate at 37°C for 20 minutes.

[0042] (6) Add 1.5 μL 20 mg / mL proteinase K and mix gently.

[0043] (7) Add 15 μL of 10% SDS, bathe in water at 50°C for 2 hours until the solution is clear, during which time gently invert and mi...

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Abstract

The invention relates to a detection application kit for Shigella sonnei wzy and wbgV gene special nucleotides. The nucleotides comprise nucleotides shown as SEQ ID NO:1 and / or nucleotides shown as SEQ ID NO:2; nucleotides shown as SEQ ID NO:3 and / or nucleotides shown as SEQ ID NO:4. The nucleotides can be used for preparing the PCR kit for detecting Shigella sonnei. The practicability for the Shigella sonnei wzy and wbgV gene special nucleotides and the PCR kit containing the nucleotides is high. The PCR kit has the characteristics of simple and convenient preparation method, short detectionperiod, high speed, high operability, high accuracy and high sensitivity, and is easy for commercial production; however, the detection cost is relatively low.

Description

technical field [0001] The present invention relates to a kind of Shigella sonnei ( Shigella sonnei ) of wzy Gene (putative polysaccharide polymerase, Putative polysaccharide polymerase, hereinafter referred to as wzy Gene ) and wxya Gene (hypothetical protein)-specific nucleotides and their application, through which Shigella sonnei can be accurately distinguished from other 13 serotypes of P. Background technique [0002] Shigella sonnei is one of the four groups of Shigella, with a single antigen and only one serotype. In my country, Shigella sonnei is the main pathogen causing bacillary dysentery. Shigella sonnei has two cross-variation phases, phase I and phase II. Phase I presents S colonies, which are pathogenic to mice and are mostly isolated from specimens of patients with acute infection. Phase II is R-type colonies, which are not pathogenic to mice and are often detected from chronic patients or carriers. [0003] At present, the typing and identificati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/10C12N15/11C12R1/01
CPCC12Q1/686C12Q1/689C12Q2563/173
Inventor 王磊经馥怿刘倩曹勃阳冯露
Owner NANKAI UNIV
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