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Primers, probe and kit for detecting enteric adenoviruses

A kit and virus technology, applied in the field of probes, kits, and primers for the detection of enteric adenoviruses, can solve problems such as inaccurate antibody detection, and achieve the effects of improving sensitivity, fast detection, and avoiding subjectivity.

Inactive Publication Date: 2018-05-08
湖北朗德医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problem of inaccurate antibody detection in the prior art, the embodiment of the present invention provides a primer, probe and kit for detecting intestinal adenovirus

Method used

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  • Primers, probe and kit for detecting enteric adenoviruses
  • Primers, probe and kit for detecting enteric adenoviruses
  • Primers, probe and kit for detecting enteric adenoviruses

Examples

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Embodiment 1

[0037] Example 1. A primer and fluorescent probe for the quantitative detection of intestinal adenovirus nucleic acid

[0038] An embodiment of the present invention provides a primer and a probe for detecting enteric adenovirus, the primers and probes include: a forward primer for detecting enteric adenovirus, a reverse primer for detecting enteric adenovirus, and a primer for detecting enteric adenovirus A probe for detecting enteric adenovirus, wherein,

[0039] The forward primer for detecting enteric adenovirus is shown in SEQ ID NO.1 in the sequence listing;

[0040] The reverse primer used to detect enteric adenovirus is shown in SEQ ID NO.2 in the sequence listing;

[0041] The probe for detecting enteric adenovirus is shown in SEQ ID NO.3 in the sequence listing;

[0042] The 5' end of the probe is connected with a fluorescent group FAM, HEX, TET, JOE, VIC, ROX, Cy3 or Cy5, and the 3' end of the probe is connected with a quencher group TAMRA, Eclipse, BHQ1, BHQ2, BH...

Embodiment 2

[0048]Example 2. A kit for detecting enteric adenovirus

[0049] The embodiment of the present invention provides a kit for quantitative detection of enteric adenovirus nucleic acid, the kit includes the primers and probes provided in Example 1 of the present invention, the kit also includes: nucleic acid release agent, PCR reaction solution, Critical Positive Controls, Positive Controls, Negative Controls, and Working Standards.

[0050] Specifically, each component of the PCR reaction solution includes in the PCR amplification reaction system: Taq enzyme with a final concentration of 0.01U / μL-0.05U / μL, dNTPs with a final concentration of 0.2-0.6mM, 10×PCR buffer and A solution containing Mg ions at a final concentration of 1.5-5.0 mM. In this example, the ratio of the contents of each component of the PCR reaction solution is: 0.3 μL of Taq enzyme at a concentration of 5 U / μL, 2 μL of dNTPs at a concentration of 10 mmol / L, 5 μL of 10×PCRBuffer, and MgCl at a concentration o...

Embodiment 3

[0084] Example 3. A kit for detecting enteric adenovirus

[0085] The embodiment of the present invention provides a kit for detecting enteric adenovirus nucleic acid in a sample. The components of the kit differ from those in the kit provided in Example 2 in that:

[0086] The ratio of the contents of each component of the PCR reaction solution is: 0.1 μL of Taq enzyme at a concentration of 5 U / μL, 1 μL of dNTPs at a concentration of 10 mmol / L, 5 μL of 10×PCR Buffer and MgCl at a concentration of 25 mmol / L 2 Solution 3 μL.

[0087] 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.

[0088] During practical application, primers and probes can be added together into the PCR reaction solution, and then sterile water can be added to a volume of 10 μL.

[0089] The DNA extraction solution included Triton-X100 with a final concentration of 0.1%, NP-40 with a final concentration of 0...

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Abstract

The invention discloses primers, a probe and a kit for detecting enteric adenoviruses, and belongs to the field of biotechnology. The primers and the probe include a forward primer for detecting the enteric adenoviruses, a reverse primer for detecting the enteric adenovirus and the probe for detecting the enteric adenoviruses. The kit comprises the primers and the probe. The primers, the probe andthe kit have the characteristic of high sensitivity, and can accurately detect whether a sample is infected by the enteric adenoviruses.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer, a probe and a kit for detecting intestinal adenovirus. Background technique [0002] Adenovirus is a common viral infection with many different clinical manifestations. Sometimes the clinical symptoms are unique and can be diagnosed clearly. However, in most cases, the symptoms caused by adenovirus are difficult to distinguish from those caused by other pathogens. Adenoviruses most commonly cause infections of the respiratory and gastrointestinal tracts. [0003] Since 1981, foreign Kidd, Takiff and others were the first to isolate enterotype adenovirus (EAD) from the feces of patients with diarrhea. Later, DeTong analyzed EAD with restriction endonuclease and found that it had two different electrophoretic patterns, named Ad- 40 and Ad-41. Adenovirus gastroenteritis is widely distributed all over the world, with a detection rate of 5% to 14%, of which Ad-40 and Ad-41 acc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2531/113C12Q2545/113C12Q2561/101
Inventor 石康朱玉华刘映乐朱世新
Owner 湖北朗德医疗科技有限公司
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