Human coronavirus nucleic acid multiplex detection kit

A coronavirus, multiplex detection technology, used in recombinant DNA technology, microbial assay/test, resistance to vector-borne diseases, etc.

Active Publication Date: 2021-05-11
上海捷诺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no method to establish a systematic, efficient, c

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  • Human coronavirus nucleic acid multiplex detection kit
  • Human coronavirus nucleic acid multiplex detection kit
  • Human coronavirus nucleic acid multiplex detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] The detection method of the human seven-fold coronavirus nucleic acid detection kit is as follows:

[0098] Samples: Various respiratory samples such as swabs, sputum, alveolar lavage fluid

[0099] Storage conditions: room temperature (18-30°C), no more than 48 hours; refrigerated temperature (2-8°C), no more than 72 hours; below -20°C, long-term storage is possible.

[0100] Steps:

[0101] 1. Extraction of sample nucleic acid.

[0102] Add 5uL internal quality control (IC) to 200uL sample, extract nucleic acid and finally elute with 100uL volume. Note that the negative control (NC) provided in the kit should be added to the internal quality control (IC) like other test samples before nucleic acid extraction. The positive control (PC) in the kit does not require nucleic acid extraction.

[0103] 2. rRT-PCR steps

[0104] 2.1 Preparation of rRT-PCR reaction solution.

[0105] Dissolve CoV detection buffer and CoV enzyme mixture at room temperature, flick to mix a...

Embodiment 2

[0125] Embodiment 2 Specificity and Sensitivity Determination

[0126] specificity test

[0127] Influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, Chlamydia pneumoniae, Mycoplasma pneumoniae, Streptococcus pneumoniae, Legionella pneumophila, Haemophilus influenzae as positive templates, cytomegalovirus, simple Herpesvirus type 1, human coronavirus NL63, human bocavirus, Epstein-Barr virus, parainfluenza virus type 1, metapneumovirus, enterovirus / rhinovirus, Mycobacterium tuberculosis, Escherichia coli, Pseudomonas aeruginosa nucleic acid is The negative template was tested by multiple detection kits for respiratory pathogens. The results showed that the detection reagents could accurately detect influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, Chlamydia pneumoniae, Mycoplasma pneumoniae, Streptococcus pneumoniae, Nucleic acid of Legionella pneumoniae and Haemophilus influenzae, and there was no false detection or miss...

Embodiment 3

[0130] Embodiment 3 clinical sample detection

[0131] Using the multiple detection system for respiratory pathogens established by the present invention, 60 cases of positive patient nucleic acid samples and 20 cases of negative samples were detected, and a commercially available single-plex PCR detection kit was used for simultaneous detection, and the positive samples were sequenced and verified .

[0132] The results show that among the 60 positive samples, commercially available single-plex fluorescent quantitative PCR and multiple fluorescent PCR methods of the present invention all detect 60 positive samples, and the positive detection rate is 100%, and 20 negative sample tests are all negative. The method consistency was high, and the results were statistically significant.

[0133] Specific to different types, the results showed 6 cases of 2019-nCoV virus, 1 case of MERS-CoV virus, 23 cases of OC43 virus, 11 cases of 229E virus, 13 cases of NL63 virus, and 6 cases of...

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Abstract

The invention provides a human coronavirus nucleic acid multiplex detection kit. Specifically, the nucleic acid RNA of seven coronaviruses, namely human coronaviruses OC43, 229E, HKU1 and NL63, middle east respiratory syndrome coronavirus (MERS-CoV), severe acute respiratory syndrome coronavirus (SARS-CoV) and acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is simultaneously detected by a multiplex rRT-PCR method, and human coronavirus nucleic acids from multiple sample sources can be rapidly detected and analyzed.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a detection kit for detecting seven kinds of human infection coronavirus nucleic acids. Background technique [0002] 2019-nCoV belongs to the genus Betacoronavirus of the Coronaviridae family, and is closely related to severe acute respiratory syndrome-related coronavirus (SARS-CoV) and Middle East respiratory syndrome-related coronavirus (MERS-CoV), both of which can cause severe pneumonia symptoms. The virus is transmitted through droplets, contact and other means. The incubation period is 3-7 days, up to 14 days. Patients in the incubation period are highly contagious. As of now, the cumulative number of confirmed cases has reached more than 20 million. Current data show that 2019-nCoV is less virulent than SARS-CoV, but more transmissible; the receptors that cause it to spread across species may be consistent with SARS-CoV, both of which are angiotensin-c...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2600/166C12Q2537/143C12Q2563/107C12Q2521/107C12Q2545/113Y02A50/30
Inventor 杨晓明秦佳华程鲁向夏小凯黄迎燕古斯·西蒙
Owner 上海捷诺生物科技有限公司
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