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Kit for quantum dot nucleic acid detection of bacterial intestinal pathogen

A technology for enteric pathogens and quantum dots, applied in the field of biomedicine, can solve the problems of small Stokes displacement, complicated operation steps, low detection sensitivity, etc., and achieve the effects of shortened detection time, low light source requirements, and high sensitivity

Pending Publication Date: 2022-04-05
杭州千基生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, gene chips mainly use organic fluorescent dyes, BCIP / NBT and DAB as detection methods. Organic fluorescent dyes have many defects: low fluorescence intensity, poor stability, easy to be photobleached, wide emission peak, small Stokes shift, BCIP / NBT and DAB methods have technical defects such as cumbersome operation steps, low detection sensitivity, and poor repeatability
[0021] It can be seen from the above-mentioned patent documents and their prior art that there has not yet been established a high-throughput, high-sensitivity, and high-specificity nucleic acid detection method for the detection of 14 bacterial strains using the optical properties of quantum dot materials and the characteristics of gene chips. Nucleic acid detection kit for enteric pathogens

Method used

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  • Kit for quantum dot nucleic acid detection of bacterial intestinal pathogen
  • Kit for quantum dot nucleic acid detection of bacterial intestinal pathogen
  • Kit for quantum dot nucleic acid detection of bacterial intestinal pathogen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Preparation and use of the kit for quantum dot nucleic acid detection of bacterial enteric pathogens of the present invention

[0095] 1. Quantum dot nucleic acid detection principle:

[0096] Molecularly hybridize the nucleic acid amplification product labeled with biotin with the probe on the detection membrane strip, and then combine biotin with quantum dots coupled with streptavidin, and observe each position of the detection membrane strip through a fluorescence detection instrument. Whether the probe is hybridized with the nucleic acid product can be judged by checking whether there is a fluorescent signal, so as to determine whether the sample contains the relevant target nucleic acid.

[0097] The capture probe is the 3' end or 5' end of the oligonucleotide single-stranded DNA labeled with an amino group, and there is an intermediate arm between the amino group and the oligonucleotide single-stranded DNA, and the intermediate arm is the fatty acid Cn chain and o...

Embodiment 2

[0208] Validation analysis of the present invention's kit for quantum dot nucleic acid detection of bacterial enteric pathogens

[0209] 1. Sensitivity detection

[0210] The reaction system was prepared according to Example 1, and divided into 21 ul, and 4 ul was added to each reaction system to extract different concentrations of pathogenic nucleic acids.

[0211] PCR amplification procedure: PCR amplification was performed according to the procedure described in Example 1.

[0212] The detection is carried out according to the kit usage procedure in Implementation 1. For test results, see figure 1 . The test results show that the sensitivity of each detection target can reach 10 3 cfu / ml.

[0213]2. Specific detection

[0214] Prepare the reaction system according to Example 1, and divide it into 21ul, add 4ul genomes to each reaction system, the detected genomes are Enterobacter cloacae, Escherichia coli, Candida albicans, Staphylococcus epidermidis, Lactobacillus bu...

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Abstract

The invention relates to the technical field of biological medicines, in particular to a kit for quantum dot nucleic acid detection of bacterial intestinal pathogens. The kit comprises a detection membrane strip, a fluorescence detection liquid and a reaction liquid, wherein the detection membrane strip comprises a nylon membrane and a capture probe fixed on the nylon membrane; the fluorescence detection liquid comprises quantum dots which are used for marking the capture probe and are coupled with streptavidin on the surface; the reaction liquid comprises a reaction liquid I, a reaction liquid II and a reaction liquid III. The kit has the beneficial effects that the kit for quantum dot nucleic acid detection of bacterial intestinal pathogens is high in throughput, high in sensitivity and high in specificity, and compared with an existing color development method gene chip, the kit has fewer steps, the detection time is obviously shortened, and meanwhile, compared with an organic fluorescent gene chip, the kit is low in equipment cost and beneficial to clinical popularization.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a kit for detecting quantum dot nucleic acid of bacterial intestinal pathogens. Background technique [0002] At present, the incidence of infectious diarrhea (ID) in my country is very high, and it is one of the important public health problems affecting human health. Infectious diarrhea (Infectious diarrhea, ID), also known as acute gastroenteritis, is a group of common intestinal infectious diseases with diarrhea and abdominal pain as the main clinical manifestations. Common pathogens of infectious diarrhea (ID) include viruses, bacteria, fungi , parasites, etc., a symptom characterized by an increase in the frequency of defecation, an increase in the amount of feces, and thin feces. Clinical manifestations include nausea, vomiting, fever, abdominal pain, tenesmus, fatigue and other symptoms, and severe cases may be accompanied by electrolyte imbalance, dehydration, renal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/6837C12N15/11
Inventor 尹华立裘惠良刘娜娜
Owner 杭州千基生物科技有限公司
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