Primers, probe and kit used for detecting BK viruses (BKVs)

A kit and virus technology, applied in the field of probes, kits, and primers for detecting BK virus, can solve the problems of slow speed and low specificity of BK virus, and achieve the effects of reliable detection results, high sensitivity and fast detection speed.

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 slow detection of BK virus and low specificity in the pri...

Method used

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  • Primers, probe and kit used for detecting BK viruses (BKVs)
  • Primers, probe and kit used for detecting BK viruses (BKVs)
  • Primers, probe and kit used for detecting BK viruses (BKVs)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

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

[0036] The reverse primer for detecting BK virus is shown in SEQ ID NO.2 in the sequence listing

[0037] The probe for detecting BK 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...

Embodiment 2

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

[0044] The embodiment of the present invention provides a kit for quantitative detection of BK virus 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.

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

Embodiment 3

[0079] Embodiment 3. A kind of kit for detecting BK virus

[0080] The embodiment of the present invention provides a kit for detecting BK virus nucleic acid in a sample. The components of the kit differ from those in the kit provided in Example 2 in that:

[0081] 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.

[0082] 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.

[0083] 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.

[0084] 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 BK viruses (BKVs), 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 BKVs. The kit comprises the primers and the probe. The primers, the probe and the kit have the characteristic of high sensitivity, the detection speed is high and the whole detection process only takes 2-3 hours.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer, a probe and a kit for detecting BK virus. Background technique [0002] BK virus (BK virus, BKV) is a kind of polyomaviruses (polyomaviruses, PV). BK virus is a non-enveloped circular double-stranded virus with a global distribution. Epidemiology shows that more than 90% of healthy adults have a history of BK virus infection in the past. BK virus reactivation occurs in patients with low immune function, and BK viruria may occur. According to reports, about 45% of kidney transplant recipients have viral activation, and 1%-10% of kidney transplant patients have BK virus associated nephropathy (BKVAN), of which 60%-70% will lead to loss of function of the transplanted kidney . In addition, BK virus infection is also associated with hemorrhagic cystitis, ureteral stricture, progressive multifocal leukoencephalopathy, lung disease, tumor, autoimmune disease, etc. in bone marr...

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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