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Rapid Campylobacter jejuni detection kit

A technology of Campylobacter jejuni and a detection kit, which is applied in the determination/inspection of microorganisms, microorganisms, biochemical equipment and methods, etc., and can solve the problems affecting the rapid diagnosis and treatment of food poisoning, easy contamination of PCR technology, false positive test results, etc. problems, to achieve the effect of fast detection speed, good specificity, and easy operation

Inactive Publication Date: 2015-03-18
SHANGHAI TONEKER BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the traditional culture method is still the main method for the detection of Campylobacter jejuni, but the operation is cumbersome and time-consuming, and it takes 4-5 days to finally determine whether it is Campylobacter jejuni, which affects the rapid diagnosis and treatment of food poisoning
With the development of molecular biology technology, PCR technology is widely used in the rapid diagnosis of bacteria, but the traditional PCR technology is easy to contaminate, resulting in false positive test results

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Reagent Composition of Campylobacter jejuni Fluorescent Quantitative PCR Detection Kit

[0038] Each kit contains 48 reactions, and the main components include:

[0039] (1) DNA extraction solution

[0040] The composition of the DNA extraction solution is: 10mM Tris-HCl, 1mM EDTA, pH=8.0, Chexe-100 with a volume fraction of 10%; a total of 5ml.

[0041] (2) PCR reaction master mix

[0042] The PCR reaction master mix includes PCR reaction buffer components (50mM KCl, 10mM Tris-HCl, pH 8.5), each 400μmol / L upstream primer hipO-1 and downstream primer hipO-2, 200μmol / L fluorescent probe hipO-3, 5mmol / L MgCl 2 , 4mmol / L dNTPs (equal volume mixture of dATP, dGTP, dCTP and dTTP) and sterile double-distilled water; a total of 2 tubes, each tube 1100ul.

[0043] (3) Hot start Taq enzyme

[0044] Hot Start Taq The concentration of the enzyme is 5U / ul; a total of 10ul.

[0045] (4) Positive standard

[0046] The positive standard is a plasmid constructed ...

Embodiment 2

[0049] Example 2: Campylobacter jejuni fluorescent quantitative PCR detection kit detects Campylobacter jejuni

[0050] The specific method is:

[0051] (1) Sample processing and DNA extraction

[0052] The method of sample processing refers to GB / T 4789.9-2008 "Food Hygiene Microbiological Examination - Campylobacter jejuni Examination".

[0053] Take 1mL of enrichment solution, centrifuge at 10 000rpm for 5 minutes, discard the supernatant to save the precipitate, add 100uL DNA extraction solution, shake and mix well; 100℃ boiling water bath or dry bath for 10 minutes; centrifuge at 10 000rpm for 5 minutes; take 2.0uL of supernatant As a template for PCR amplification, the rest were stored at -20°C. (If the extracted supernatant has not been subjected to PCR amplification on the same day, it is recommended to store it at -20°C for no more than 1 month; before using it again, it should be centrifuged at 10 000 rpm for 5 minutes.)

[0054] (2) Prepare PCR reaction solution ...

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PUM

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Abstract

The invention relates to a fluorescent quantitative PCR (polymerase chain reaction) detection kit for rapidly detecting Campylobacter jejuni DNA. The kit is suitable for qualitatively or quantitatively detecting the Campylobacter jejuni DNA; and the fluorescent quantitative Campylobacter jejuni PCR detection kit comprises a DNA extract liquid, a PCR premix liquid, a hot start Taq enzyme, a positive standard substance and a negative quality control. Compared with traditional bacterium culture methods and biochemical identification, the fluorescent quantitative Campylobacter jejuni PCR detection kit has the advantages of strong specificity, high sensitivity, good repeatability, quantification accuracy, fast speed, high flux, time saving, simple operation and the like.

Description

technical field [0001] The invention relates to a fluorescent quantitative PCR (polymerase chain reaction) detection kit for rapidly detecting the presence of Campylobacter jejuni DNA, which is suitable for qualitative or quantitative detection of the presence of Campylobacter jejuni DNA. Background technique [0002] Campylobacter jejuni (Campylobacter jejuni) is one of the most common foodborne pathogens causing gastrointestinal infection bacterial enteritis. Common pathogenic bacteria, most poultry and livestock carry this bacteria, parasitize in their intestines, feces or intestinal contents after slaughter spread in the surrounding environment, and then contaminate meat or other foods, causing food poisoning. It can cause acute gastroenteritis, and can also cause systemic diseases such as endocarditis, arthritis, sepsis, and thrombotic veins, and is also closely related to the occurrence of acute infectious polyradiculoneuropathy in humans, namely Guillain-Barré syndro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/01
CPCC12Q1/6851
Inventor 刘代新潘海云
Owner SHANGHAI TONEKER BIOTECH