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Respiratory tract pathogen rapid fluorescence PCR detection kit and primer probe combination thereof

A detection kit and primer probe technology, applied in the field of biotechnology applications, can solve the problems of low detection throughput, long detection period, low detection sensitivity of respiratory fever pathogens, etc., and achieve high detection sensitivity, good accuracy, and throughput. big effect

Inactive Publication Date: 2019-01-15
南通国际旅行卫生保健门诊部
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a rapid fluorescent PCR detection kit for respiratory pathogens and its combination of primers and probes to solve the problems of low detection sensitivity, long detection cycle and low detection throughput of respiratory fever pathogens in the prior art

Method used

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  • Respiratory tract pathogen rapid fluorescence PCR detection kit and primer probe combination thereof
  • Respiratory tract pathogen rapid fluorescence PCR detection kit and primer probe combination thereof
  • Respiratory tract pathogen rapid fluorescence PCR detection kit and primer probe combination thereof

Examples

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Embodiment 2

[0035] Example 2 Respiratory tract infectious disease detection operation specification

[0036] 1. Nucleic acid extraction

[0037]1.1 Nucleic acid extraction: The kit is equipped with DNA / RNA one-step lysis and extraction reagents. Use 200 μL serum or throat swab sample to add 50ul lysate, centrifuge at 5000RPM for 2min after 95℃ for 2min, and take 50μL supernatant as a template. For specific extraction steps, please refer to the corresponding extraction Kit instructions.

[0038] 2. Reagent preparation

[0039] 2.1 According to the number of samples to be tested (n), take out the corresponding number of 16-well chip plates, negative and positive control reagents, and briefly centrifuge after melting at room temperature;

[0040] 2.2 Move the above-mentioned 16-well chip plate to the sample processing area.

[0041] 3. Adding samples

[0042] Use a suction nozzle with a filter element to take 50 μL of the processed sample supernatant and add them to the sample wells of t...

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Abstract

The invention discloses a rapid fluorescent PCR detection kit for respiratory tract pathogens and a primer probe combination thereof. The kit comprises an amplification chip and a control reagent. Thecontrol reagent comprises a negative control and a positive control. The detection reagent comprises RT-PCR reaction solution consisting of Buffer, 2.0 mM dNTPs, 1 U / [mu]m L Taq DNA polymerase, 2 U / [mu]m L M-MLV reverse transcriptase, 0.3 U / [mu]m L RRI and 16 primer probes for detection of 9 respiratory pathogens. Sixteen types of primer probe mixtures of nine respiratory pathogens are pre-loadedinto a 16-channel microfluidic amplification tube by the method of reagent freeze-thawing intervention, Combined with the rapid microfluidic real-time fluorescent PCR amplification instrument, 16 types of infectious diseases associated with respiratory tract fever symptoms can be screened at one time.

Description

technical field [0001] The invention discloses a rapid fluorescent PCR detection kit for respiratory pathogens and a primer-probe combination thereof, belonging to the application field of biotechnology. Background technique [0002] Respiratory tract infection is one of the most common diseases in the world, and the incidence rate occupies a major position in the overall structure of the morbidity rate of residents in various countries. At the peak of the epidemic each year, about 10% of the residents suffer from respiratory tract infection. Respiratory tract infections are mainly caused by various respiratory viruses and some bacteria, mycoplasma and chlamydia. Common viruses are influenza A virus, influenza B virus, parainfluenza virus, coronavirus, respiratory syncytial virus, human metapneumovirus, human boca virus, respiratory adenovirus, rhinovirus, etc. Most of these respiratory viruses are highly infectious and spread quickly, with a short incubation period and acu...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/701C12Q2600/16C12Q2537/143C12Q2563/107C12Q2531/113C12Q2545/101C12Q2565/629C12Q2561/101Y02A50/30
Inventor 陈峰吴海磊杨庆贵杨国平
Owner 南通国际旅行卫生保健门诊部
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