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Primers, probe and kit used for detecting JC viruses (JCVs)

A JC virus and kit technology, applied in the field of probes, kits, and primers for detecting JC virus, can solve problems such as inaccurate antibody detection, and achieve the effects of improving sensitivity, high sensitivity, and reliable detection results.

Inactive Publication Date: 2015-07-01
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 of inaccurate antibody detection in the prior art, the embodiment of the present invention provides a primer, probe and kit for detecting JC virus

Method used

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  • Primers, probe and kit used for detecting JC viruses (JCVs)
  • Primers, probe and kit used for detecting JC viruses (JCVs)
  • Primers, probe and kit used for detecting JC viruses (JCVs)

Examples

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Effect test

Embodiment 1

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

[0034] The embodiment of the present invention provides a primer and a probe for detecting JC virus. The primer and probe include: a forward primer for detecting JC virus, a reverse primer for detecting JC virus and a primer for detecting JC virus. probes, where,

[0035] The forward primer used to detect JC virus is shown in SEQ ID NO.1 in the sequence listing;

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

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

[0038] 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, BHQ3 or DABCYL . In the embodiment of the present invention, the fluorescent reporting group of the probe is FAM, the e...

Embodiment 2

[0043] Embodiment 2. A kind of kit for detecting JC virus

[0044] The embodiment of the present invention provides a kit for detecting the quantitative nucleic acid of JC virus. The kit includes the primers and probes provided in Example 1 of the present invention. Positive controls, positive controls, negative controls, and working standards.

[0045]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 with a concentration of 5 U / μL, 2 μL of dNTPs with a concentration of 10 mmol / L, 5 μL of 10×PCR Buffer and 25 μL of dNTPs with a concentration of 25 mmol / L MgCl 2 Solution 5 μL.

[0046] Specifically, the final co...

Embodiment 3

[0078] Embodiment 3. A kind of kit for detecting JC virus

[0079] The embodiment of the present invention provides a kit for detecting JC virus nucleic acid in a sample. The components of the kit differ from those in the kit provided in Example 2 in that:

[0080] 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.

[0081] 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.

[0082] 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.

[0083] The DNA extraction solution included Triton-X100 with a final concentration of 0.1%, NP-40 with a final concentration of 0.2% and T...

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Abstract

The invention discloses primers, probe and kit used for detecting JC viruses (JCVs), belonging to the technical field of biology. The primers and probe comprise a forward primer, a reverse primer and a probe, which are used for detecting JCVs. The kit comprises the primers and the probe. The primers, the probe and the kit have the characteristic of high sensitivity and can be used for accurately detecting whether samples are infected with the JCVs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer, a probe and a kit for detecting JC virus. Background technique [0002] JC virus (JC virus, JCV) belongs to the Papovaviridae polyomavirus species human polyomavirus branch. In 1971, Pagett et al first isolated JC virus from the brain tissue of patients with progressive multifocal leucoencephalopathy (PML). JC virus exists widely in the human population, mainly lurking in tissues such as human kidneys. The infection of JC virus occurs in childhood, and it is in a state of continuous subclinical latent infection. In a healthy immune state, it can be asymptomatic for life, which belongs to asymptomatic infection. However, for AIDS patients or patients taking immunosuppressive drugs, it can cause PML and other series PML is a fatal demyelinating disease of the central nervous system. In recent years, studies have shown that JC virus is associated with the occurrence of a va...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6844C12Q1/70C12Q2563/107
Inventor 刘茹桑筱筱
Owner 湖北永邦医疗科技有限公司
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