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Kit and method for detecting food-borne pathogenic bacteria

A food-borne pathogenic bacteria and detection kit technology, applied in the field of molecular biology, can solve the problems of cumbersome operation steps, inability to cope with pathogenic bacteria, and long time consumption

Inactive Publication Date: 2011-09-14
SHENZHEN INT TRAVEL HEALTHCARE CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional detection of food-borne pathogens mainly adopts the method of bacterial culture, which is more accurate in detection results, but it takes a long time and the operation steps are cumbersome; in addition, real-time fluorescent PCR method can also be used for detection, which is specific strong, but one experiment can only detect one pathogenic target, and it cannot cope well with the detection of a large number of pathogenic bacteria at the same time
These two methods often take a long time to detect a variety of foodborne pathogens, which also delays the best time for diagnosis and treatment

Method used

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  • Kit and method for detecting food-borne pathogenic bacteria
  • Kit and method for detecting food-borne pathogenic bacteria
  • Kit and method for detecting food-borne pathogenic bacteria

Examples

Experimental program
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Effect test

Embodiment 1

[0102] Design and prepare primers and probes

[0103] By analyzing the gene sequences of 12 common food-borne pathogenic bacteria, a relatively conservative sequence is selected as the target amplification sequence, and the PCR for amplifying the target amplification sequence is designed according to the target amplification sequence Amplification primers and probes that specifically bind to the targeted amplification sequence.

[0104] The 12 targeted amplification sequences and their corresponding 12 sets of PCR amplification primers and 12 probes are as follows:

[0105] The first targeted amplification sequence is the nucleic acid fragment shown in SEQ ID NO.1 of Staphylococcus aureus, the corresponding upstream primer sequence is shown in SEQ ID NO.13, and the downstream primer sequence is shown in SEQ ID NO.25. Shown, the probe sequence is shown in SEQ ID NO.37.

[0106] The second targeted amplification sequence is the nucleic acid fragment shown in SEQ ID NO.2 of Sal...

Embodiment 2

[0121] Foodborne pathogenic bacteria detection kit and detection method

[0122] The detection kit includes the above-mentioned 12 sets of primers and 12 probes.

[0123] In a preferred embodiment, the detection kit also includes a PCR positive quality control probe, a PCR positive detection probe complementary to the PCR positive quality control probe, and a PCR positive detection probe primer for amplifying the PCR positive detection probe Pair, positive hybridization control probe, and positive hybridization detection probe complementary to the positive hybridization control probe.

[0124] The PCR positive quality control probe sequence is shown in SEQ ID NO.49, the PCR positive detection probe sequence is shown in SEQ ID NO.50, and the upstream primer sequence of the PCR positive detection probe primer pair is shown in SEQ ID NO.51 , the downstream primer sequence of the PCR positive detection probe primer pair is shown in SEQ ID NO.52, the sequence of the hybridization ...

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Abstract

The invention relates to a kit for detecting food-borne pathogenic bacteria, which comprises 12 groups of primers and probes thereof, wherein in the primers, the 5' ends of downstream primers are all tagged with biotin. The kit amplifies nucleic acid in a sample to be detected by utilizing the primers, and a formed biotinylated product and a probe fixed on the surface of a chip are hybridized to form a probe-biotinylated product compound; an alkaline phosphatase-streptavidin conjugate is combined with the probe-biotinylated product compound, and then chromogenic substrate which is tetranitroblue tetrazolium chloride / 5-bromine-4-chlorine-3-indolyl-phosphate (NBT / BCIP) is reacted with alkaline phosphatase to undergo a colour generation (bluish violet) reaction; and whether the sample to be detected contains the pathogenic bacteria to be detected is detected by interpreting whether a specific site has colour generation. The kit has the advantages of high detection throughput, high speed, high sensitivity, high specificity and great practicability. The invention also provides the method for detecting the food-borne pathogenic bacteria by adopting the detection kit.

Description

【Technical field】 [0001] The invention relates to the field of molecular biology, in particular to a detection kit and detection method for food-borne pathogenic bacteria. 【Background technique】 [0002] Humans eat a wide variety of foods, and there are also various pathogens transmitted through food, including bacteria, viruses, rickettsia, toxin-producing algae, parasites, and large fungi such as mushrooms. In daily life, food poisoning incidents caused by bacteria, especially food-borne pathogenic bacteria-contaminated food are the most common. [0003] The traditional detection of food-borne pathogenic bacteria mainly adopts the method of bacterial culture, which is more accurate in detection results, but it takes a long time and the operation steps are cumbersome; in addition, real-time fluorescent PCR method can also be used for detection, which is specific However, one experiment can only detect one pathogenic target, and it cannot cope well with the detection of a l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCY02A50/30
Inventor 顾大勇李永进张巍史蕾赵芳刘春晓赵纯中杨燕秋徐云庆孙杰莫秋华杨泽庄卫东
Owner SHENZHEN INT TRAVEL HEALTHCARE CENT
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