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Coronavirus detection product and application thereof

A coronavirus and product technology, applied in the field of molecular biology, can solve the problems of increasing the time for patients to seek medical treatment, high cost of patient testing, missed detection of coronavirus, etc., and achieve the effect of reducing medical treatment time, easy operation and high sensitivity

Active Publication Date: 2020-12-25
SHANGHAI ZJ BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There are currently two methods most commonly used in clinical practice for the detection of coronaviruses: one is for suspected patients, first detect whether they are infected with other influenza viruses, and if they are not infected with influenza viruses, continue to screen for other items, such as new coronaviruses (or Influenza virus) detection, this method is time-consuming and labor-intensive, which not only increases the time for patients to seek medical treatment, but also easily causes the risk of missed detection of other coronaviruses, such as patients with other coronavirus infections, or the selected detection gene loci A new type of coronavirus has mutated; the second is for suspected patients, multiple items are tested at one time to screen for pathogens, but this method is too expensive for patients to detect, and the problem of missed detection is still unresolved

Method used

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  • Coronavirus detection product and application thereof
  • Coronavirus detection product and application thereof
  • Coronavirus detection product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0121] Embodiment 2 The present invention compares with other commercialization extraction kits extraction effect

[0122] 1. Experimental method

[0123] 1.1 Detection object

[0124] Construct a high-concentration Coronavirus simulated positive sample. The simulated positive sample is composed of the synthetic product of the new coronavirus genome sequence (as shown in SEQ ID NO.10 and SEQ ID NO.11) and the background matrix of normal human exfoliated cells. Take a high-concentration Coronavirus simulated sample. For positive samples, three gradients were diluted with Coronavirus-negative nasopharyngeal swab wash samples and normal saline, respectively. The three concentration samples diluted in the coronavirus negative nasopharyngeal swab wash sample are respectively defined as XL-1, XL-2, and XL-3, and the three concentration samples diluted in normal saline are respectively defined as XN1, XN2, and XN3.

[0125] 1.2 Inactivation of samples

[0126] Take the above sampl...

Embodiment 3

[0150] Application of the Coronavirus joint detection kit in embodiment 3 saliva and sputum

[0151] 1. Experimental method

[0152]1.1 Detection object

[0153] The saliva and sputum obtained from 4 clinical cases were tested for Coronavirus nucleic acid. These 4 samples have been identified as 2 Coronavirus positive samples and 2 Coronavirus negative samples by culture method in the hospital, which are numbered S1, S2, S3 and S4 in sequence.

[0154] 1.2 Sample inactivation and nucleic acid extraction

[0155] Take the above samples and add 1.5mL virus inactivator respectively, shake and mix well, and let stand at room temperature for 30min.

[0156] Use the self-contained extraction reagent of the present invention to carry out the extraction of 4 examples of saliva and sputum samples (same as 1.3 in Example 2). Add 1 μL of internal standard to row A of the preloaded plate, and then add S1, S2, S3, S4, negative quality control or positive quality control to each well in ...

Embodiment 4

[0165] Coronavirus detection kit performance evaluation in embodiment 4 sputum

[0166] 1. Experimental method

[0167] 1.1 Detection object

[0168] (1) Sensitivity analysis

[0169] Take 1 copy of Coronavirus simulated positive sample, dilute it 10 times with sample diluent to form 5 gradients, numbered S1, S2, S3, S4 and S5 in sequence, add negative quality control and positive quality control, nucleic acid extraction and detection .

[0170] The detection method is as follows: Take 30 μL of the PCR detection mixture, mix it with 0.5 μL of PCR enzyme, shake and mix for a few seconds, centrifuge at 3000 rpm for a few seconds, take 30 μL of the above-mentioned mixed liquid and put it in a PCR tube, and then add 20 μL of the sample, negative quality control or Add the nucleic acid extraction product of the positive quality control product into the PCR reaction tube, cover the PCR reaction tube cap, and proceed to the PCR amplification reaction immediately.

[0171] The cyc...

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Abstract

The invention relates to the technical field of molecular biology, in particular to a coronavirus detection product and an application thereof. A detection substance comprises a primer and probe for specifically recognizing coronavirus nucleic acid, the primer comprises an upstream primer and a downstream primer, and the upstream primer is selected from a first upstream primer as shown in SEQ ID NO: 1 or a second upstream primer as shown in SEQ ID NO: 2; the downstream primer is selected from a first downstream primer as shown in SEQ ID NO: 3 or a second downstream primer as shown in SEQ ID NO: 4; and the probe is selected from a first probe as shown in SEQ ID NO: 5 or a second probe as shown in SEQ ID NO: 6. According to the coronavirus detection product, the current situation that no detection reagent for the oropharyngeal swabs, the nasopharyngeal swabs, the saliva, the sputum, the excrement and the blood samples exists is solved for the first time, coronavirus total viruses are qualitatively detected, and missing detection of other coronaviruses except the new coronaviruses is prevented.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a coronavirus detection product and its application. Background technique [0002] Coronaviruses belong to the genus Coronavirus of the family Coronaviridae in systematic classification. Viruses of the genus Coronavirus are positive-sense single-stranded RNA viruses with an envelope (envelope), and their genetic material is the largest among all RNA viruses. Different types of viruses have different pathogenicity and cause different clinical manifestations. The symptoms caused by the OC43 strain are generally more severe than those caused by the 229E virus. Coronavirus is one of the main pathogens of common cold in adults, and the incubation period of infection is generally 2-5 days. Typical coronavirus infection shows cold symptoms such as runny nose and malaise, and symptoms such as fever, chills, and vomiting may also occur. The representative variants are mainly ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/118C12Q2600/166C12Q2561/101C12Q2561/113C12Q2563/107
Inventor 李强史艳梅张阳明雷永兴王占坤张捷邵俊斌
Owner SHANGHAI ZJ BIO TECH
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