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Staphylococcus aureus enterotoxin gene PCR (Polymerase Chain Reaction) parting detection kit and method

A staphylococcus entero and detection kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of undetected Staphylococcus aureus enterotoxin, etc.

Inactive Publication Date: 2011-02-16
长沙市疾病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no genotyping detection kit and method for detecting Staphylococcus aureus enterotoxin SEA, SEB, SEC, SEE by PCR technology at present

Method used

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  • Staphylococcus aureus enterotoxin gene PCR (Polymerase Chain Reaction) parting detection kit and method
  • Staphylococcus aureus enterotoxin gene PCR (Polymerase Chain Reaction) parting detection kit and method
  • Staphylococcus aureus enterotoxin gene PCR (Polymerase Chain Reaction) parting detection kit and method

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Embodiment 1

[0047] Make PCR-based Genotyping Detection Kit for Staphylococcus aureus Enterotoxin according to the following formula:

[0048] (1) Amplification reaction solution components:

[0049] 10×Taq DNA Polymerase Buffer, 250mM dNTP, ddH 2 O: 10×Taq DNA Polymerase Buffer contains 200mM trishydroxymethionine methane-hydrochloride pH 8.4, 200mM potassium chloride, 100mM ammonium sulfate, 15mM magnesium sulfate and 1% Triton X-100.

[0050] (2) Typing reaction solution A: Contains 20uM each of SEA upstream and downstream primers:

[0051] SEA-F: 5′-CCGAGTCACGATCAATTTTTACAG-3′

[0052] SEA-R: 5'-GCGGCTTGTATATAAATATATATCA-3';

[0053] (3) Typing reaction solution B: Contains 20uM each of SEB upstream and downstream primers:

[0054] SEB-F: 5′-AATCTATAGATCAATTTCTATAC-3′

[0055] SEB-R: 5'-CTTTTTCTTTGTCGTAAGATA-3';

[0056] (4) Typing reaction solution C: Contains 20uM each of SEC upstream and downstream primers:

[0057] SEC-F: 5′-GTCTGTAGATAAATTTTTGGCA-3′

[0058] SEC-R: 5'-TCCA...

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Abstract

The invention relates to a gene parting detection kit for detecting staphylococcus aureus enterotoxin SEA, SEB, SEC and SEE by a PCR (Polymerase Chain Reaction) method as well as a method thereof, belonging to the field of biological detection reagents. The kit comprises amplification reaction liquid, parting reaction liquids A, B, C and E, Taq enzyme, ddH2o, positive standard product of SEA, SEB, SEC and SEE, wherein the parting reaction liquid A contains SEA primers: SEA-F and SEA-R; the parting reaction liquid B contains SEB primers: SEB-F and SEB-R; the parting reaction liquid C contains SEC primers: SEC-F and SEC-R; the parting reaction liquid E contains SEE primers: SEE-F and SEE-R. The detection method comprises the steps of: extracting bacteria DNA, performing parting detection on the enterotoxin SEA, SEB, SEC and SEE genes by using the parting primers designed in the invention under the same PCR instrument and the same condition, and displaying the result via electrophoresis. The detection is rapid, accurate, high in sensitivity and low in cost.

Description

technical field [0001] The invention belongs to the field of biological detection reagents, and relates to a genotyping detection kit for detecting Staphylococcus aureus enterotoxins SEA, SEB, SEC and SEE by PCR method, and using the kit to rapidly detect Staphylococcus aureus enterotoxins Methods for SEA, SEB, SEC, SEE genes. Background technique [0002] Staphylococcus aureus (SA) is one of the most common bacterial food poisoning pathogens at home and abroad. Among the poisoning outbreaks caused by it, the "Snowprint Milk Powder" incident in 2000 was the first in recent years, and more than 14,000 people were infected. . According to figures released by the Ministry of Health of Japan, there were 2,525 outbreaks of food poisoning caused by Staphylococcus aureus during the 20-year period (1980-1999) in Japan, infecting 59,964 people. The main cause of food poisoning caused by this bacteria is the production of staphylococcal enterotoxins (SEs). According to its serologic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/02
Inventor 孙边成张如胜欧新华宋克云陈法明
Owner 长沙市疾病预防控制中心
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