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Primer probe composition, kit containing primer probe composition and detection method of primer probe composition

A primer-probe and kit technology, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc., can solve the problems of low detection efficiency, poor specificity and low sensitivity of the kit, and achieve accurate detection. High sensitivity and sensitivity, accurate results and high detection efficiency

Active Publication Date: 2022-02-18
GUANGZHOU KEFEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The second object of the present invention is to provide a joint detection kit for detecting influenza A virus, influenza B virus and respiratory syncytial virus nucleic acid, which can realize multiple inspections in one tube, and completely solve the low detection efficiency of the kit in the prior art , poor specificity and low sensitivity

Method used

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  • Primer probe composition, kit containing primer probe composition and detection method of primer probe composition
  • Primer probe composition, kit containing primer probe composition and detection method of primer probe composition
  • Primer probe composition, kit containing primer probe composition and detection method of primer probe composition

Examples

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Effect test

Embodiment 1

[0046] The primer-probe composition for detecting influenza A virus, influenza B virus and respiratory syncytial virus, and the preparation of the kit comprising it are as follows:

[0047] The primer and probe sequences in the kit are shown in Table 1 below:

[0048] Table 1 Primer, probe sequence set

[0049]

[0050] In this embodiment, the 5' end of the probe shown in SEQ ID NO:3 is marked with a VIC fluorescent group, and the 3' end is marked with a BHQ1 quenching group, so as to ensure that influenza A viruses can be distinguished during the reaction and improve detection efficiency.

[0051] In this embodiment, the 5' end of the probe shown in SEQ ID NO: 6 is marked with a ROX fluorescent group, and the 3' end is marked with a BHQ2 quenching group to ensure that influenza B viruses can be distinguished during the reaction and improve detection efficiency.

[0052]In this embodiment, the 5' end of the probe shown in SEQ ID NO: 9 is marked with a FAM fluorescent gro...

Embodiment 2

[0057] Methods for detecting influenza A virus, influenza B virus, and respiratory syncytial virus for non-diagnostic purposes

[0058] The detection method adopted in the present invention is q-PCR, and the PCR reaction solution, DNA polymerase, reverse transcriptase and sample nucleic acid are prepared into a nucleic acid amplification reaction mixture according to the proportion of the following reaction system for PCR reaction. The proportioning ratio of the reaction system is shown in Table 2:

[0059] Table 2 Configuration of nucleic acid amplification reaction mixture

[0060]

[0061] After the nucleic acid amplification reaction mixture is configured, the PCR reaction is carried out according to the procedure shown in Table 3:

[0062] Table 3 Fluorescent quantitative PCR reaction conditions

[0063]

[0064] Selection of fluorescence detection channels: select VIC channel to detect influenza A virus nucleic acid; select ROX channel to detect influenza B virus...

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Abstract

The invention discloses a primer probe composition, a kit containing the primer probe composition and a detection method of the primer probe composition, belonging to the technical field of virus detection. The primer probe composition comprises specific primers and probes aiming at the influenza A virus, the influenza B virus and the respiratory syncytial virus. The specific primer and the probe have good specificity. High-specificity, high-sensitivity and high-efficiency detection on the influenza A virus, the influenza B virus and the respiratory syncytial virus can be realized by adopting the kit and a Q-PCR method.

Description

technical field [0001] The invention belongs to the technical field of virus detection, and in particular relates to a primer probe composition for detecting influenza A virus, influenza B virus and respiratory syncytial virus, a kit containing the same and a detection method thereof. Background technique [0002] Respiratory tract infection (RTI) is the most common disease in humans and one of the leading causes of morbidity and mortality worldwide. The clinical manifestations caused by respiratory tract infection are mainly rhinitis, pharyngitis, laryngitis, tonsillitis and other symptoms, and severe cases can cause tracheitis, bronchitis and pneumonia, etc. However, the treatment methods, curative effects and course of the infection caused by different pathogens are also different. same. It has been proven that most of the respiratory infections are caused by respiratory viruses. Common respiratory viruses include influenza A virus, influenza B virus and respiratory sync...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 李娇娇
Owner GUANGZHOU KEFEN BIOTECH CO LTD
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