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Kit for quantum dot nucleic acid detection of urinary tract infection-causing pathogens

A urinary tract infection and quantum dot technology, applied in the field of biomedicine, can solve the problems of complex preparation process, unfavorable promotion, complicated and cumbersome detection process, etc., and achieve high specificity, high throughput, and high sensitivity

Active Publication Date: 2018-10-16
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, DAB and organic fluorescence as detection methods. Organic fluorescent dye gene chips have many defects: the preparation process is complex, the detection process is complex and cumbersome, and the detection equipment requires expensive laser scanners , not conducive to clinical promotion
[0028] It can be seen from the above-mentioned patent documents and other prior art that there is currently no method for establishing a high-throughput, high-sensitivity, high-specificity nucleic acid detection method for urinary tract infection by utilizing the optical characteristics of quantum dot materials and the characteristics of gene chips. Detection kit

Method used

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  • Kit for quantum dot nucleic acid detection of urinary tract infection-causing pathogens
  • Kit for quantum dot nucleic acid detection of urinary tract infection-causing pathogens
  • Kit for quantum dot nucleic acid detection of urinary tract infection-causing pathogens

Examples

Experimental program
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Effect test

Embodiment 1

[0113] Preparation and use of the kit for quantum dot nucleic acid detection of urinary tract infection pathogens according to the present invention

[0114] 1. Quantum dot nucleic acid detection principle:

[0115] 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.

[0116] 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 dT (n) ...

Embodiment 2

[0237] Validation analysis of the present invention's kit for quantum dot nucleic acid detection of urinary tract infection pathogens

[0238] 1. Sensitivity detection

[0239] The genomic DNA of each pathogenic bacteria at the calibration concentration was serially diluted (10pg / μL, 1pg / μL, 0.5pg / μL, 0.1pg / μL, 0.05pg / μL, 0.01pg / μL), and passed the PCR-quantum dot fluorescence detection method The above concentrations of DNA were tested separately, and those with obvious bright spots were judged as positive results, and those without spots were judged as negative results. The detection is carried out according to the kit usage procedure in Implementation 1. For test results, see figure 1 .

[0240] 2. Specific detection

[0241] The reaction system was prepared according to the above-mentioned Example 1, and divided into 21 ul, and 4 ul of genomic DNA was added to each reaction system. The detected genomic DNA was Streptococcus pyogenes, Streptococcus agalactiae, Streptoco...

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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 urinary tract infection-causing pathogens. The kit comprises a detection membrane strip, a fluorescent detection solution and reaction solutions, wherein the detection membrane strip comprises a nylon membrane and a capture probe fixed to the nylon membrane; the fluorescent detection solution comprises quantum dots for marking the capture probe and coupled with streptavidin on the surfaces; the reaction solutions include a reaction solution I, a reaction solution II, a reaction solution III and a reaction solution IV. The kit has the beneficial effects as follows: the high-throughput, high-sensitivity and high-specificity kit for the quantum dot nucleic acid detection of the urinary tract infection-causing pathogens is provided; the kit has fewer steps and obviously shorter detection time than the existing colorimetric gene chip, has lower equipment cost than an organic fluorescent gene chip, and is conducive 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 urinary tract infection pathogens. Background technique [0002] Urinary tract infection (Urinary Tract Infection, UTI) refers to the inflammation caused by pathogenic microorganisms invading the urinary system. Its condition recurs, protracted, and can range from asymptomatic bacteriuria to severe toxemia. It can occur in various ages and genders, and is common in women. Its incidence is relatively high, second only to infectious diseases of the respiratory tract and digestive tract. The main clinical manifestations are bladder irritation symptoms such as frequent urination, urgency, and painful urination. Some patients also have symptoms such as fever and suprapubic tenderness. If the treatment is not timely, it can also cause complications such as bacteremia, acute or chronic renal insufficiency, etc. symptom. [0003] Pat...

Claims

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Application Information

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IPC IPC(8): C12Q1/6837C12Q1/6895C12Q1/689C12Q1/14C12Q1/10C12Q1/04
CPCC12Q1/6837C12Q1/689C12Q1/6895C12Q2600/16C12Q2565/519C12Q2563/131C12Q2563/107C12Q2565/625C12Q2531/113C12Q2537/143
Inventor 尹华立唐小利郑银娜裘惠良
Owner 杭州千基生物科技有限公司
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