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Respiratory tract pathogen detecting kit

A pathogen detection and kit technology, applied in the field of molecular biology, can solve problems such as inability to identify pathogen species, and achieve the effects of reducing the risk of cross-contamination, high specificity and low cost

Inactive Publication Date: 2015-11-04
JIANGSU PROVINCIAL CENT FOR DISEASE PREVENTION & CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when only multiplex PCR technology is used to identify different respiratory pathogens, the size of nucleic acid fragments amplified by different pathogens needs to be significantly different, otherwise the pathogen species cannot be identified

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: 9 kinds of common respiratory pathogen kit detection methods

[0023] 1 Sample selection: select common respiratory pathogens: RNA viruses, including H3N2, H1N1, swH1N1, H5N1, FluB, SARS-COV viruses. DNA viruses or bacteria, including LP, N.men and ADV viruses. Virus strains or samples are isolated, identified and preserved in our laboratory.

[0024] 2 Nucleic acid extraction: Nucleic acid extraction was carried out in accordance with the requirements of the Roche MagNA Pure LC Automatic Nucleic Acid Extractor and MagNA Pure LC Total Nucleic Acid Isolation Kit, and the nucleic acid was extracted and stored at -80°C.

[0025] 3 reverse transcription: according to Invitrogen SuperScript First-Strand Synthesis SuperMix manual operation method, use the random primers provided by the kit to reverse-transcribe FluA, FluB, SARS-COV viral nucleic acid into DNA and store at -20°C.

[0026] 4. Design and synthesis of multiplex PCR primers and nucleic acid invasion ...

Embodiment 2

[0035] Embodiment 2 Using the kit of the present invention to detect the specificity of common respiratory pathogens

[0036] Use the optimized multiplex PCR system to amplify nucleic acid samples of FluA, FluB, SARS-COV, LP, N.men, ADV, and human metapneumovirus. color method for detection. The results are shown in Table 3. Respiratory pathogens: FluA, FluB, SARS-COV, LP, N.men, and ADV can only be detected with corresponding primer probes when using multiplex PCR reaction technology combined with nucleic acid invasion reaction and nano-gold color development. When the nano-gold develops color, it will show positive results, and the results are all negative when detecting other pathogens, indicating that this method has high specificity for detecting respiratory pathogens.

[0037] Table 3 The kit of the present invention detects the specific detection results of common pathogens in the respiratory tract

[0038]

[0039] Note: √ indicates that the test result is positiv...

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Abstract

The invention discloses a respiratory tract pathogen detecting kit. The kit is used on the basis of multiple PCR combining nucleic acid intrusion reaction and the nano-gold color developing principle. The kit comprises one or more of 6 primer probe combinations and can be used for detecting influenza A / avian influenza viruses (FluA), influenza B viruses (FluB), SARS coronaviruses, legionella pneumophila (LP), neisseria meningitidis (N. men) and adenoviruses (ADV). The kit has the advantages that the kit is high in specificity and sensitivity during respiratory tract pathogen detecting, expensive equipment is not needed, detecting results can be observed through naked eyes, and the kit can be applied at primary level.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a respiratory pathogen detection kit. Background technique [0002] Respiratory tract infection is one of the most common diseases in humans. Some common respiratory pathogens such as influenza / avian influenza virus, adenovirus, Legionella pneumophila and Neisseria meningitidis are the most common infections in young children, the elderly, the infirm and immunocompromised main cause of morbidity and mortality. The infection of these pathogens has the characteristics of strong infectivity, fast transmission, short incubation period, acute onset, and no lifelong immunity after illness. Health and safety pose a great hazard. [0003] Because the infection symptoms and epidemic characteristics caused by respiratory pathogens are similar, and it is difficult to determine the true pathogens only by clinical symptoms, and the multi-virus co-infection, virus mutation and the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12Q1/04C12R1/93C12R1/36C12R1/01
CPCC12Q1/689C12Q1/701
Inventor 樊欢戚宇华朱政崔仑标葛以跃史智扬吴涛
Owner JIANGSU PROVINCIAL CENT FOR DISEASE PREVENTION & CONTROL
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