Detection of real-time fluorescent polyase chain reaction of respiratory pathogen

A chain reaction and real-time fluorescence technology, which is applied in the determination/examination of microorganisms, biochemical equipment and methods, etc., can solve the problems that cannot meet clinical needs and cannot meet the needs of early pathogen diagnosis.

Active Publication Date: 2006-03-15
GUANGZHOU HUAYIN MEDICINE SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

Since it takes a certain amount of time for pathogens to enter the body for replication and antigen stimulation to produce antibodies, this method still cannot meet the needs of early diagnosis of pathogens, and thus cannot meet the clinical needs of early active treatment

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1: The real-time fluorescent PCR assay method and kit for respiratory pathogens of the present invention.

[0067] 1 kit composition:

[0068] (1) DNA extraction reagent: including A solution: 6mol / L guanidine isothiocyanate, 30mmol / L sodium citrate, 0.58% sodium dodecylsulfonate, 1mmol / L disodium edetate, 2% Glycogen; liquid B: isopropanol; liquid C: 75% ethanol in water; sample diluent: ddH 2 O.

[0069] (2) PCR reaction reagents: including PCR reaction solution and enzymes for each pathogen.

[0070] For RSV, EV, the PCR reaction solution of Influenza.A or Influenza.B (20 μ l per person, add the pathogen template 5 μ l that extracts during detection, the final volume of reaction solution is 25 μ l) components are as follows: 5 μ l of 5 × RT reaction solution ( 250mmol / L Tris-HCl, pH 9.2, 250mmol / L KCl, 20mmol / L MgCl 2 , 50mmol / L DTT); final concentration of 0.3mmol / L dNTP; 0.12μmol / L probe; 0.6μmol / L nested PCR inner primer; 0.06μmol / L nested PCR outer pr...

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PUM

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Abstract

The invention is about the PCR detecting method of a respiratory tract pathogen and the reagent box. The character is to design the vector and the probe according to the highly special base groups of the pathogen by analyzing the nucleic acid sequence of the pathogen. So next to amplify the PCR or the RT-PCR in the fluorescent PCR instrument.

Description

technical field [0001] The invention relates to a real-time fluorescent polymerase chain reaction (PCR) detection method and a kit for pathogens of seven respiratory infectious diseases. These seven pathogens include: Influenza A virus (Influenza.A), Influenza B virus (Influenza.B), Respiratory syncytial virus (RSV), Enterovirus (EV), Mycoplasma pneumoniae (MP), Chlamydia pneumoniae ( CP), respiratory adenovirus (ADV). Background technique [0002] Respiratory pathogens invade the human body mainly through the respiratory tract, and spread outward with respiratory secretions, and invade another susceptible body to cause respiratory infectious diseases. Respiratory pathogens are mainly transmitted through the air (droplets, dust), but there is still a lack of effective large-scale air disinfection methods, so the focus of prevention is on early detection and pre-epidemic prevention. Respiratory pathogens mainly include viruses, pathogenic bacteria, mycoplasma, chlamydia, et...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 周荣黄茜华王海
Owner GUANGZHOU HUAYIN MEDICINE SCI & TECH
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