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Detection kit for novel coronavirus, influenza A and B and respiratory syncytial virus

A technology of syncytial virus and respiratory tract, which is applied in the fields of biomedicine and clinical diagnosis, can solve the problems of many co-infections of patients, adverse effects of new coronavirus prevention and control, and great hidden dangers of cross-infection, so as to reduce the burden on hospitals, false negatives and false negatives. Low positive, low false positive effect

Inactive Publication Date: 2020-05-01
GUANGZHOU HYBRIBIO MEDICINE TECH LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If fever cases cannot be diagnosed quickly and accurately in a timely manner, influenza cases are likely to be treated as new coronavirus infections, and there are many patients with co-infection and high risk of cross-infection, which will have an extremely adverse impact on the prevention and control of new coronaviruses.

Method used

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  • Detection kit for novel coronavirus, influenza A and B and respiratory syncytial virus
  • Detection kit for novel coronavirus, influenza A and B and respiratory syncytial virus
  • Detection kit for novel coronavirus, influenza A and B and respiratory syncytial virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Primer and probe design and screening

[0055] 1. The present invention analyzes several viral sequences and explores through a large number of studies, and designs primers and probe sets shown in Table 1-4:

[0056] Table 1 Combination of primers and probes of the present invention

[0057]

[0058] Table 2 Control group 1

[0059]

[0060] Table 3 Control group 2

[0061]

[0062] Table 4 Control group 3

[0063]

[0064] 2. At the same time, after optimization and exploration of the corresponding detection system, the following detection system was established: 23.5 μl RT-PCR amplification solution and 1.5 μl enzyme mixture were added to each PCR tube. And add 5 μl template RNA amount.

[0065] That is, the RT-PCR detection system (total volume 30μl) includes:

[0066] (1) 23.5 μl RT-PCR amplification solution, including primers, probes and PCR buffer;

[0067] The concentration of the probe used to detect the target gene was 200 nM, the c...

Embodiment 2

[0082] Example 2 kit

[0083] 1. New coronavirus, influenza A virus, influenza B virus and respiratory syncytial virus nucleic acid joint detection kit, including the following components:

[0084] (1) Primer and probe combination

[0085]By analyzing the DNA of clinical samples, the inventors obtained the primers and SEQ ID NOs of SEQ ID NO: 1~2, as shown in Table 1, which have complementary nucleotide sequences only to the 2019-nCoV ORF1ab of the new coronavirus, as shown in Table 1. The probe of ID NO: 3; the primer of SEQ ID NO: 4 ~ 5 and the probe of SEQ ID NO: 6 that are only complementary to the nucleotide sequence of the 2019-nCoV N target gene; only complementary to influenza A virus The nucleotide sequence of SEQ ID NO: 7 ~ 8 primers and the probe of SEQ ID NO: 9; only the primers and SEQ ID NO of the complementary nucleotide sequence of influenza B virus: 10 ~ 11 : the probe of 12; only with the primer of SEQ ID NO: 13~14 and the probe of SEQ ID NO: 15 of the comp...

Embodiment 3

[0111] Example 3 Detection example

[0112] 1. Experiment

[0113] In order to test the accuracy and effectiveness of the kit of the present invention for the detection of the new coronavirus 2019-nCoV, influenza A, influenza B and respiratory syncytial virus, we tested the quality control products of the A-fluid plasmid, the B-fluid plasmid quality control, Respiratory syncytial virus plasmid quality control, new crown plasmid quality control, internal standard plasmid quality control (different concentrations: 1×10 6 copies / ml, 1×10 5 copies / ml, 1×10 4 copies / ml, 1×10 3 copies / ml, 1×10 2 copies / ml, 1×10 1 copies / ml) for testing, and Guangdong CDC new crown external quality assessment quality control product 5330 (dilute different concentrations: 1×10 5 copies / ml, 1×10 4 copies / ml, 1×10 3 copies / ml, 1×10 1 copies / ml), 18 cases of quality control products from Bondeson Company were tested.

[0114] 2. Results

[0115] The result is as Figure 1-6 The amplification ...

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Abstract

The invention discloses a detection kit for a novel coronavirus, influenzas A and B and a respiratory syncytial virus. According to the detection kit, through combining primer probe design and adaptive detection system optimizing research, a group of nucleic acid combined detection primer and probe combinations and nucleic acid combined detection kits for the novel coronavirus (2019-nCoV / SARS-CoV2), an influenza A virus (FluA), an influenza B virus (FluB) and the respiratory syncytial virus (RSV) are developed, and sequences of primers and probes are shown in SEQ ID NO: 1-18. The kit has theadvantages of high detection sensitivity, good repeatability and low false negative and false positive and can effectively distinguish novel coronavirus infection from ordinary viral influenza and respiratory syncytial virus induced respiratory tract infection, so that precise diagnosis and subsequent precise treatment on a sufferer are achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and clinical diagnosis. More specifically, it relates to a nucleic acid joint detection kit for novel coronavirus (2019-nCoV / SARS-CoV-2), influenza A virus (FluA), influenza B virus (FluB) and respiratory syncytial virus (RSV) . Background technique [0002] Since December 2019, the new coronavirus 2019-nCoV / SARS-CoV-2 is spreading in the world, but humans lack effective antiviral vaccines and specific drugs. Currently, accurate detection and diagnosis of infected persons or carriers is of great importance. Epidemic control has a crucial role and significance. In addition, it is now a period of high incidence of various influenza A virus, influenza B virus, respiratory syncytial virus and other related virus infections. Their clinical manifestations are similar to those of the new coronavirus pneumonia, and it is difficult to distinguish them through clinical manifestations and chest imaging...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q2600/166C12Q1/6848C12Q2531/113Y02A50/30
Inventor 朱娟娟李烈军谢龙旭
Owner GUANGZHOU HYBRIBIO MEDICINE TECH LTD
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