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Detection kit and detection method for Campylobacter jejuni in pork

A technology of Campylobacter jejuni and a detection kit, which is applied in the direction of microorganism-based methods, microorganism measurement/inspection, biochemical equipment and methods, etc., can solve the problems of inaccurate detection results, shorten the detection time, and reduce false positives. Negative rate, simple operation effect

Inactive Publication Date: 2015-01-14
中山鼎晟生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Denaturing high-performance liquid chromatography (DHPLC) uses the principle of reversed-phase high-performance liquid chromatography to separate nucleotide fragment molecules. The operation is fully automated, with high accuracy and sensitivity. However, when the content of Campylobacter jejuni in food is extremely low, the detection results not very accurate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A detection kit of campylobacter jejuni in pork, described detection kit comprises substratum and detection liquid; Described substratum is broth; Described detection liquid is made of the component of following content:

[0042] 10PCR Buffer 3μl,

[0043] Taq DNA polymerase 0.2μl,

[0044] dNTP 3 μl,

[0045] 100bp DNA Ladder Marker 0.1μl,

[0046] PCR primer 1μl,

[0047] Tris.Cl 5mg,

[0048] Sodium chloride 1mg,

[0049] Magnesium Chloride 1mg,

[0050] Disodium hydrogen phosphate dodecahydrate 0.2mg,

[0051] Enrichment solution 3 mg.

[0052] Described broth is Bolton broth, contains the growth promoter of 0.3% weight portion in the broth; Described growth promoter is sodium metabisulfite, sodium pyruvate and ferrous sulfate mixed in the weight portion of 2:1:5 mixture.

[0053] The PCR primers are: C16S-F: 5'-CTG CTT AAC ACA AGT TGA GTA GG-3'(183~205), C16S-R: 5'-TTC CTT AGG TAC CGT CAG AA-3'(449 ~468).

[0054] The Taq DNA polymerase concentration is 2U...

Embodiment 2

[0063] A detection kit of campylobacter jejuni in pork, described detection kit comprises substratum and detection liquid; Described substratum is broth; Described detection liquid is made of the component of following content:

[0064] 10PCR Buffer 5μl,

[0065] Taq DNA polymerase 0.4μl,

[0066] dNTPs 5 μl,

[0067] 100bp DNA Ladder Marker 0.6μl,

[0068] PCR primer 2μl,

[0069] Tris.Cl 10mg,

[0070] Sodium Chloride 2.2mg,

[0071] Magnesium Chloride 2.2mg,

[0072] Disodium hydrogen phosphate dodecahydrate 0.5mg,

[0073] Enrichment solution 5 mg.

[0074] Described broth is Bolton broth, contains the growth promoter of 0.5% weight portion in the broth; Described growth promoter is sodium metabisulfite, sodium pyruvate and ferrous sulfate mixed in the weight portion of 2:1:5 mixture.

[0075] The PCR primers are: C16S-F: 5'-CTG CTT AAC ACA AGT TGA GTA GG-3'(183~205), C16S-R: 5'-TTC CTT AGG TAC CGT CAG AA-3'(449 ~468).

[0076] The Taq DNA polymerase concentrati...

Embodiment 3

[0085] A detection kit of campylobacter jejuni in pork, described detection kit comprises substratum and detection liquid; Described substratum is broth; Described detection liquid is made of the component of following content:

[0086] 10PCR Buffer 4μl,

[0087] Taq DNA polymerase 0.3μl,

[0088] dNTP 4 μl,

[0089] 100bp DNA Ladder Marker 0.5μl,

[0090] PCR primer 1.5μl,

[0091] Tris.Cl 8mg,

[0092] Sodium chloride 2mg,

[0093] Magnesium Chloride 1.5mg,

[0094] Disodium hydrogen phosphate dodecahydrate 0.3mg,

[0095] Enrichment solution 4 mg.

[0096] Described broth is Bolton broth, contains the growth promoter of 0.4% weight portion in the broth; Described growth promoter is sodium metabisulfite, sodium pyruvate and ferrous sulfate mixed in the weight portion of 2:1:5 mixture.

[0097] The PCR primers are: C16S-F: 5'-CTG CTT AAC ACA AGT TGA GTA GG-3'(183~205), C16S-R: 5'-TTC CTT AGG TAC CGT CAG AA-3'(449 ~468).

[0098] The Taq DNA polymerase concentration...

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Abstract

The invention relates to a detection kit for Campylobacter jejuni in pork. The detection kit comprises a culture medium and a detection solution, wherein the culture medium is broth. The detection solution is composed of 3-5 mu l of 10 PCR (polymerase chain reaction) buffer solution, 0.2-0.4 mu l of Taq DNA (deoxyribonucleic acid) polymerase, 3-5 mu l of dNTP (deoxyribonucleotide triphosphate), 0.1-0.6 mu l of 100bp DNA Ladder Marker, 1-2 mu l of PCR primer, 5-10 ml of Tris.cl, 1-2.2mg of sodium chloride, 1-2.2mg of magnesium chloride, 0.2-0.5mg of disodium hydrogen phosphate dodecahydrate and 3-5mg of enrichment broth. The detection kit is an improvement on the basis of the conventional PCR technology, and can be used for qualitative detection of Campylobacter jejuni. By integrating the advantages of PCR, DHPLC (denaturing high performance liquid chromatography) and other techniques, the detection kit has the advantages of high specificity of detection results and high sensitivity, and ensures the effective detection of Campylobacter jejuni in food.

Description

technical field [0001] The invention belongs to the field of food testing, and in particular relates to a detection kit and a detection method for Campylobacter jejuni in pork. Background technique [0002] Pork is the main meat species in people's daily life. Whether the content of microorganisms in pork reaches the standard is directly related to everyone's dietary health. Campylobacter jejuni is one of the most susceptible microorganisms in pork products. It is the main cause of foodborne bacterial gastroenteritis in humans and animals worldwide, and can cause acute enteritis and diarrhea. Reports show that Campylobacter jejuni infection accounts for 47% of all foodborne bacterial infections, and can cause sporadic and Endemic gastroenteritis outbreaks, especially malignant tumors, AIDS, chronic diseases, children and the elderly have the highest incidence. In addition, it can also cause Guillain-Barré syndrome (GBS), reactive arthritis, endocarditis, etc. A variety of ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/01
CPCC12Q1/6848C12Q2531/113C12Q2565/137
Inventor 郭狄
Owner 中山鼎晟生物科技有限公司
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