Primer sequences and quantitative determination kit used for simultaneously detecting human CMV and BK virus DNA

A quantitative detection and virus technology, applied in the direction of recombinant DNA technology, DNA / RNA fragments, microbial measurement / inspection, etc., to achieve the effect of multi-experimental information, simple composition, and reduced manpower and material resources
CN102344970BInactive Publication Date: 2013-01-23THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL COLLEGE
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL COLLEGE
Publication Date
2013-01-23
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses primer sequences and a quantitative determination kit used for simultaneously detecting human CMV and BK virus DNA. The invention discloses primers, fluorescent probes, and a real-time quantitative polymerase chain reaction (real-time PCR) kit used for simultaneously carrying out quantitative determinations on deoxyribonucleic acid (DNA) segments of cytomegalovirus (CMV) and BK virus (BKV). The kit comprises reagent A, B, C, D, E, and G. The reagent A is composed of a mixed set of primers and probes, wherein the primers and probes are targeted at consensus sequences ofCMV DNA and BKV DNA. The reagent B is a buffer solution containing dNTP. The reagent C is Tag DNA polymerase. The reagent D is composed of standard samples with different concentrations. The reagent E and G are respectively positive and negative controls. With the whole set of kit provided by the invention, quantitative determinations can be simultaneously carried out on DNA of CMV and BK virusesin clinical specimens of urine, blood or other body fluids.
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Description

technical field

[0001] The present invention relates to biotechnology, is a kind of molecular biology method based on quantitative nucleic acid amplification, particularly relates to using real-time fluorescent quantitative polymerase chain reaction (real-time PCR), by using specific primers to amplify cytomegalovirus (CMV ) and polyomavirus (BKV) conserved fragments, and quantify them by releasing fluorescent signals from specific probes. After optimizing the experimental conditions, a kit that can simultaneously detect CMV DNA and BKV DNA in a single reaction tube was constructed. Background technique

[0002] CMV and BKV are latent viruses in kidney transplant recipients. When immunosuppressive therapy is overdone, both viruses reactivate to varying degrees and damage the transplanted kidney. These viruses can be found in the urine, plasma, and peripheral blood cells of kidney transplant recipients, and have a huge negative impact on the long-term survival of kidney tra...

Claims

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