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Primer, probe and kit all used for detecting mumps virus

A mumps virus and kit technology, applied in the field of kits, can solve problems such as the low positive rate of ELISA kits and cannot meet emergency diagnosis, and achieve reliable detection results, improved sensitivity, and high sensitivity

Inactive Publication Date: 2015-05-06
湖北永邦医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that the low positive rate of the ELISA kit in the prior art cannot meet the needs of emergency diagnosis, the embodiment of the present invention provides a primer, probe and kit for detecting mumps virus

Method used

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  • Primer, probe and kit all used for detecting mumps virus
  • Primer, probe and kit all used for detecting mumps virus
  • Primer, probe and kit all used for detecting mumps virus

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Example 1. A primer and fluorescent probe for the quantitative detection of mumps virus nucleic acid

[0034] The embodiment of the present invention provides a primer and probe for detecting mumps virus, the primer and probe include: a forward primer for detecting mumps virus, a reverse primer for detecting mumps virus and a A probe for detecting mumps virus, wherein,

[0035] The forward primer for detecting mumps virus is shown in SEQ ID NO.1 in the sequence listing;

[0036] The reverse primer used to detect mumps virus is shown in SEQ ID NO.2 in the sequence listing;

[0037] The probe for detecting mumps virus is shown in SEQ ID NO.3 in the sequence listing;

[0038] The 5' end of the probe is connected with fluorescent groups FAM, HEX, JOE, TET, Cy3, Cy5, ROX, Fluorescein, Rhodamine, Rhodamine Red, Rhodamine 6G, Orengon Green 488, Orengon Green 500, Orengon Green 514, Texas Red, TAMRA, Inosine, FITC or Acridine orange, the 3' end of the probe is connected with...

Embodiment 2

[0043] Example 2. A kit for detecting mumps virus

[0044]The embodiment of the present invention provides a kit for quantitative detection of mumps virus nucleic acid, the kit includes the primers and probes provided in Example 1 of the present invention, and the kit also includes: RNA extraction solution, RT-PCR reaction solution , Working Standards, Positive Controls, Borderline Positive Controls, and Negative Controls.

[0045] Specifically, the RT-PCR reaction solution includes: Taq enzyme at a final concentration of 0.01U / μL-0.05U / μL, reverse transcriptase at a final concentration of 0.4U / ul-2U / ul, dNTPs, 10×PCR buffer and a solution containing Mg ions at a final concentration of 1.5-5.0 mM. The ratio of the content of each component of the specific RT-PCR reaction solution is: 0.3 μL of Taq enzyme at a concentration of 5 U / μL, 0.3 μL of reverse transcriptase at a concentration of 200 U / μL, 2 μL of dNTPs at a concentration of 10 mmol / L, 10× PCR Buffer 5μL and MgCl at a...

Embodiment 3

[0074] Example 3. A kit for detecting mumps virus

[0075] The embodiment of the present invention provides a kit for detecting Streptococcus pneumoniae caused by Streptococcus mumps in a sample, the difference between the components of the kit and the kit provided in Example 2 is that:

[0076] The ratio of the contents of each component in the RT-PCR reaction solution is: 0.1 μL of Taq enzyme at a concentration of 5 U / μL, 0.1 μL of reverse transcriptase at a concentration of 200 U / μL, 1 μL of dNTPs at a concentration of 10 mmol / L, and 10×PCR Buffer5μL and MgCl at a concentration of 25mmol / L 2 Solution 3 μL.

[0077] Add 0.25 μL each of forward primer and reverse primer with a concentration of 10 μmol / L, and add 0.25 μL of probe with a concentration of 10 μmol / L.

[0078] In practical application, primers and probes can be added together into the RT-PCR reaction solution, and then sterile water can be added to a volume of 10 μL.

[0079]The RNA extraction solution included...

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Abstract

The invention discloses a primer, a probe and a kit all used for detecting mumps virus, and belongs to the technical field of kits. The primer and the probe comprise a forward primer for detecting mumps virus, a reverse primer for detecting mumps virus and a probe for detecting mumps virus. The kit comprises the primer and the probe. The primer, the probe, the kit and a detection method possess the characteristic of high sensitivity, the detection speed is fast, and the whole detection process only needs 2-3 h.

Description

technical field [0001] The invention relates to the field of kits, in particular to a primer, a probe and a kit for detecting mumps virus. Background technique [0002] Mumps is an acute respiratory infectious disease caused by mumps virus (MuV) infection. Mumps is often overlooked because of its mild clinical symptoms and is a self-limiting disease. However, the disease is often accompanied by various complications. The virus can invade multiple organs and the central nervous system, resulting in a variety of clinical symptoms. Common complications include viral meningitis, encephalitis, orchitis, epididymal Inflammation, oophoritis, pancreatitis, myocarditis, etc., can lead to disability or death in severe cases. [0003] Mumps is relatively common clinically, and its diagnosis is mainly based on clinical manifestations. In addition, ELISA (Enzyme Linked Immunosorbent Assay, enzyme-linked immunosorbent assay) method can also be used to detect mumps virus IgM antibody kit...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/701
Inventor 刘茹桑筱筱
Owner 湖北永邦医疗科技有限公司
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