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Method for detecting respiratory tract pathogens in to-be-detected sample and kit and application thereof

A pathogen and respiratory technology, which is applied in the field of detection of respiratory pathogens in samples to be tested, can solve the problems of long cultivation period, inability to quickly detect pathogens in samples in a short time, low detection sensitivity and specificity, etc.

Pending Publication Date: 2021-09-07
GUANGDONG FAPON BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the bacteria and virus culture method has a long culture cycle and a harsh culture environment, which cannot meet the requirements of rapid detection of sample pathogens in a short period of time, and may delay the best time for treatment; although antibody immunoassays can detect multiple pathogens at the same time, their detection is limited. It is the specific antibody produced by the body caused by the invasion of pathogens in the sample to be tested, but the production of specific antibody takes time, there is a long window period, and the detection sensitivity and specificity are also low, prone to false positives
[0005] PCR usually includes gel band analysis method and fluorescence detection method, among which gel band analysis method has low sensitivity, and blurred bands may lead to errors in judgment, resulting in misdiagnosis; fluorescence detection method usually uses fluorescent dyes or fluorescent probes as the base Markers that enable signal recognition for singleplex or multiplex PCR

Method used

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  • Method for detecting respiratory tract pathogens in to-be-detected sample and kit and application thereof
  • Method for detecting respiratory tract pathogens in to-be-detected sample and kit and application thereof
  • Method for detecting respiratory tract pathogens in to-be-detected sample and kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] This embodiment provides a reagent for detecting respiratory pathogens in a sample to be tested, which includes a first reagent to a fourth reagent, as shown in Table 1 in detail.

[0102] Table 1 sequence

[0103]

[0104]

[0105] Note: "F" in Table 1 refers to the upstream primer, and "R" refers to the downstream primer.

[0106] The target regions of the first to fourth reagents in Table 1 are target regions 1a to 4a, respectively.

Embodiment 2

[0108] This embodiment provides a kit for diagnosing respiratory-related diseases, which includes the first to fourth reagents provided in Example 1, RT-PCR buffer, enzyme mixture and positive quality control.

[0109] Wherein, RT-PCR buffer includes: including PCR buffer, dATP, dUTP, dTTP, dCTP, dGTP and MgCl 2 ;

[0110] Enzyme Mix includes: Hot Start DNA Polymerase, Reverse Transcriptase, RNase Inhibitor and UNG Enzyme;

[0111] Positive quality control products include quality control products 1-4, please refer to Table 2 for the target sequences contained in quality control products 1-4.

[0112] Table 2 Target sequence

[0113]

[0114]

[0115] .

Embodiment 3

[0117] This embodiment provides a method for detecting respiratory pathogens in a sample to be tested, and the method includes the following steps.

[0118] Using the kit provided in Example 2, the samples to be tested were subjected to multiplex fluorescent PCR amplification according to the following system and conditions.

[0119] The final concentration of the primer pair 1 in the amplification system is 750nM, and the final concentration of the probe 1 in the amplification system is 1000nM;

[0120] The final concentration of the primer pair 2 in the amplification system is 750nM, and the final concentration of the probe 2 in the amplification system is 1000nM;

[0121] The final concentration of the primer pair 3 in the amplification system is 400nM, and the final concentration of the probe 3 in the amplification system is 500nM;

[0122] The final concentration of the primer pair 4 in the amplification system is 750 nM, and the final concentration of the probe 4 in the...

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Abstract

The invention discloses a method for detecting respiratory pathogens in a to-be-detected sample and a kit and application thereof, and relates to the technical field of biology. The method comprises the step of detecting a to-be-detected sample with a first reagent for detecting influenza A virus, a second reagent for detecting influenza B virus, a third reagent for detecting mycoplasma pneumoniae and / or a fourth reagent for detecting chlamydia pneumoniae, and a new way is provided for effectively detecting respiratory pathogens or simultaneously detecting multiple respiratory pathogens.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting respiratory pathogens in a sample to be tested, a kit and application thereof. Background technique [0002] Respiratory system disease is a common and frequently-occurring disease. The main lesions are in the trachea, bronchi, lungs, and chest cavity. Mild cases often cause coughing, chest pain, and respiratory problems. Severe cases have dyspnea, hypoxia, and even death due to respiratory failure. It is one of the major diseases threatening human health. [0003] The pathogens that cause respiratory diseases mainly include viruses, bacteria, mycoplasma, chlamydia, and rickettsia, etc., but the clinical symptoms caused by different pathogens are very similar. Therefore, when diagnosing pathogens of respiratory diseases, misdiagnosis or missed diagnosis is easy , which in turn leads to inappropriate use of drugs (such as antibiotics). [0004] Traditional det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/689C12Q1/6851C12Q1/04C12N15/11C12R1/93C12R1/35
CPCC12Q1/701C12Q1/689C12Q1/6851C12Q2531/113C12Q2563/107
Inventor 毛平道林钰琼
Owner GUANGDONG FAPON BIOTECH CO LTD
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