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Respiratory pathogen multiplex detection kit and application thereof

A detection kit and respiratory technology, applied in the field of molecular biology, can solve problems such as complex operation process, manual on-duty, and long detection time, and achieve the effects of simplifying experimental operation steps, preventing environmental pollution, and improving detection throughput

Pending Publication Date: 2022-04-08
无锡科智达科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In addition, in the existing technology, the extraction and detection of nucleic acid are usually carried out in sections, the operation process is complicated, and a professional molecular detection laboratory is required. The detection takes a long time and costs high, which cannot meet the diverse detection needs and requires manual guarding. and participation, which brings a lot of inconvenience to the nucleic acid extraction and detection process

Method used

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  • Respiratory pathogen multiplex detection kit and application thereof
  • Respiratory pathogen multiplex detection kit and application thereof
  • Respiratory pathogen multiplex detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Respiratory pathogen multiple detection kit, said kit includes specific primers and fluorescent probes for amplifying five gene loci of influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus and mycoplasma pneumoniae; Among them, the 5 loci are: M gene of influenza A virus, NS gene of influenza B virus, M gene of respiratory syncytial virus, H gene of adenovirus, Mgp gene of Mycoplasma pneumoniae; and human reference gene RP specific primers and fluorescent probes.

[0023] Preferably, the sequences of the specific primers and fluorescent probes are as follows: FluA, SEQ ID NO.1-3; FluB, SEQ ID NO.4-6; RSV, SEQ ID NO.7-9; ADV, SEQ ID NO. 10-12; MP, SEQ ID NO. 13-15; Homo, SEQ ID NO. 16-18.

[0024] Table 1 Specific primers and fluorescent probe sequence list

[0025]

[0026]

[0027] Preferably, the kit includes PCR reaction solution I and PCR reaction solution II; wherein, PCR reaction solution I includes SEQ ID NO.1-2, SEQ ID NO.4-5, SE...

Embodiment 2

[0032] The influenza virus ribonucleic acid (IVARNA) liquid indoor quality control product, the influenza A H5N1 influenza virus ribonucleic acid (H5N1RNA) liquid indoor quality control product, the influenza A H7N9 influenza virus ribonucleic acid (H7N9RNA) liquid indoor quality control product, H9N2 influenza virus ribonucleic acid (H9N2RNA) liquid indoor quality control material, influenza B virus ribonucleic acid (IVBRNA) liquid indoor quality control material, respiratory syncytial virus type B ribonucleic acid liquid indoor quality control material, human metapneumovirus RNA ( HMPVRNA) liquid indoor quality control, adenovirus deoxyribonucleic acid (ADVDNA) liquid indoor quality control, adenovirus type 11 deoxyribonucleic acid (ADV11DNA) liquid indoor quality control, mycoplasma pneumoniae deoxyribonucleic acid (MPDNA) liquid indoor quality control Add human parainfluenza type 1 virus ribonucleic acid (PIV1RNA) liquid indoor quality control product, human parainfluenza t...

Embodiment 3

[0038] Embodiment 3 Sensitivity detection test

[0039] Dilute the plasmid DNA containing the respective targets of influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus and mycoplasma pneumoniae with a certain number of copies in sterilized purified water, and then add them to the detection wells of the detection device on the detection.

[0040] It can be shown from the detection results that the detection sensitivity of the detection method for influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus and Mycoplasma pneumoniae is 500 copies / reaction.

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Abstract

The invention discloses a multiplex detection kit for respiratory pathogens and application of the multiplex detection kit. The kit comprises specific primers and fluorescent probes for amplifying five gene loci of influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus and mycoplasma pneumoniae. Compared with the prior art, the invention has the following advantages: (1) the multiple fluorescent quantitative PCR amplification technology is combined with the freeze drying technology, simultaneous detection of multiple indexes of a single sample is realized, the system simplifies experimental operation steps, improves the detection flux, provides a new method for accurate detection of multiple indexes of five respiratory pathogens, and has good application prospects. The urgent requirements of multi-index rapid screening and accurate detection are met; (2) in the detection process, the detection areas are completely isolated from each other and sealed, and after detection is completed, the sealing reagent is cured, so that outward diffusion of amplification products in the whole detection process and after detection is completed is effectively prevented, and environmental pollution is prevented.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to the detection of five kinds of human respiratory pathogens, in particular to a multiple detection kit for respiratory pathogens and an application thereof. Background technique [0002] Acute respiratory infection is a common infectious disease that is widely distributed in the population, has high morbidity and mortality among the elderly and infants, and brings serious economic burden to the society. Acute upper respiratory tract infection and lower respiratory tract infection can be caused by bacteria, virus and mycoplasma infection, mainly caused by respiratory virus. The reasons are: virus particles are small and easy to spread through the respiratory tract; viruses are prone to mutations and produce new highly pathogenic mutants; at the same time, with the development of new technologies, new pathogens are constantly being discovered. Among the viruses that cause r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/689C12Q1/6851C12N15/11C12R1/93C12R1/35
Inventor 陈俊飞邓永杰项春燕唐大运刘梅
Owner 无锡科智达科技有限公司
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