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Kit for detecting quantum dot nucleic acid of bloodstream infection pathogen

A technology of quantum dots and kits, applied in the field of biomedicine, can solve the problems of small Stokes shift, cumbersome operation steps, and poor repeatability

Active Publication Date: 2018-10-09
杭州千基生物科技有限公司 +1
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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
[0023] It can be seen from the above-mentioned patent literature and its prior art that there is currently no high-throughput, high-sensitivity, high-specificity nucleic acid detection method for bloodstream infection that utilizes the optical characteristics of quantum dot materials and the characteristics of gene chips. Detection kit

Method used

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  • Kit for detecting quantum dot nucleic acid of bloodstream infection pathogen
  • Kit for detecting quantum dot nucleic acid of bloodstream infection pathogen
  • Kit for detecting quantum dot nucleic acid of bloodstream infection pathogen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] Preparation and use of the kit for quantum dot nucleic acid detection of bloodstream infection pathogens of the present invention

[0121] 1. Quantum dot nucleic acid detection principle:

[0122] 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 hybridizes with the nucleic acid product is determined by spotting the light signal, so as to determine whether the sample contains the relevant target nucleic acid.

[0123] The capture probe is the 3' 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 a fatty acid C(n) chain or oligo Combination of dT(n...

Embodiment 2

[0259] Effect verification analysis of the kit for quantum dot nucleic acid detection of bloodstream infection pathogens according to the present invention

[0260] 1. Sensitivity detection

[0261] The reaction system was prepared according to Example 1, and divided into 21 ul, and 4 ul of genomic DNA was added to each reaction system, and the concentration of genomic DNA was 10 pg / ul, 1 pg / ul, and 0.1 pg / ul.

[0262] PCR amplification procedure: PCR amplification was performed according to the procedure described in Example 1.

[0263] The detection is carried out according to the kit usage procedure in Implementation 1. For test results, see figure 1 . The detection results show that the sensitivity of each detection target can reach 0.1pg / ul.

[0264] 2. Specific detection

[0265] The reaction system was prepared according to Example 1, and divided into 21ul, and 4ul of genomic DNA was added to each reaction system. The detected genomic DNA was the genomic DNA of Can...

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Abstract

The invention relates to the technical field of biological medicines, specifically to a kit for detecting quantum dot nucleic acid of a bloodstream infection pathogen. The kit comprises a detecting membrane strip, a fluorescence detecting solution and a reaction solution, wherein the detecting membrane strip comprises a nylon membrane and a capture probe fixed to the nylon membrane; the fluorescence detecting solution comprises quantum dots for marking streptavidin which is coupled on the surface of the capture probe; the reaction solution comprises a reaction solution I, a reaction solution II, a reaction solution III and a reaction solution IV. The kit has the beneficial effects of being high in throughput, flexibility and specificity; the steps are fewer than the steps of the conventional color-developing gene chip; the detection time is obviously shortened; the cost is less than that of an organic fluorescence gene chip device; the kit is beneficial for 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 bloodstream infection pathogens. Background technique [0002] Blood stream infection (Blood Stream Infection, BSI) mainly includes sepsis and bacteremia. Septicemia is a blood infection caused by various pathogenic microorganisms (bacteria or fungi) and toxins invading the bloodstream. The main clinical manifestations are: sudden chills, high fever, tachycardia, shortness of breath, rash, hepatosplenomegaly A series of serious clinical symptoms such as Yamato spirit and mental changes, severe cases can cause shock, disseminated intravascular coagulation (DIC) and multiple organ failure. If the bacteria only invade human blood for a short time without clinically obvious symptoms of toxemia (such as blood vessel-related infection), it is called bacteremia. Bloodstream infections (BSIs) are often critical and are one of the lead...

Claims

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Application Information

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IPC IPC(8): C12Q1/6834C12Q1/689C12Q1/04C12N15/11
CPCC12Q1/6834C12Q1/689C12Q2600/16C12Q2600/166C12Q2537/143C12Q2545/101C12Q2531/113C12Q2563/107
Inventor 尹华立张爽郑银娜裘惠良
Owner 杭州千基生物科技有限公司
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