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Fluorescent PCR detection method for human cytomegalovirus (HCMV)

A technology of human cytomegalovirus and detection method, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of pollution, long operation steps and detection time, and difficult to popularize, so as to reduce pollution and detect The effect of short time and lower false positive rate

Inactive Publication Date: 2010-06-30
SHANGHAI FOSUN PHARMA (GROUP) CO LTD +1
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AI Technical Summary

Problems solved by technology

But at the same time, the operation steps and detection time are long, and the experiment is easy to cause pollution, so it is difficult to carry out extensive clinical promotion

Method used

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  • Fluorescent PCR detection method for human cytomegalovirus (HCMV)
  • Fluorescent PCR detection method for human cytomegalovirus (HCMV)

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Embodiment Construction

[0044] Design and prepare primers and probe sequences (designed for the HCMV standard strain-Human herpesvirus 5strain AD169 gene sequence design):

[0045] Primer 1: 5`-ACCCTGTTTCTTCCTCGCTAT-3` (Seq No.1)

[0046] Primer 2: 5`-GCCCGAGCCCGACTTTA-3` (Seq No.2)

[0047] Primer 3: 5`-CCAGTGAATTTTCTCTTCCGTCT-3` (Seq No.3)

[0048] Primer 4: 5`-CATAGAGATAGCGATAAATGAGTC-3` (Seq No.4)

[0049] Probe 1: 5`FAM-GATACAGCCGGCGGTATCGA-BHQ1-3` (Seq No.5)

[0050] Probe 2: 5`FAM-CTCCTTGATTCTATGCCGCACCAT-BHQ1-3`(Seq No.6)

[0051] The above primers and probes were all synthesized in Bao Biological Engineering (Dalian) Co., Ltd.

[0052] The human cytomegalovirus (HCMV) fluorescent PCR diagnostic kit consists of (20 servings):

[0053] Ingredient Name Volume

[0054] Nucleic acid extraction solution 1ml

[0055] PCR reaction buffer 460μl

[0056] PCR reaction enzyme 40μl

[0057] Positive control substance 50μl

[0058] Negative control substance 50μl

[0059] Among them, the PCR r...

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Abstract

The invention relates to a fluorescent PCR detection method for a human cytomegalovirus (HCMV). In the invention, two groups of detecting primers and probes are designed for UL122 and UL134 genes of a human cytomegalovirus (HCMV) genome. By detecting the two genes simultaneously, a false negative rate caused by the mutation of the HCMV genome is reduced. The technology of the invention can perform qualitative and quantitative detection on HCMV DNA quickly and conveniently and has relatively high specificity and sensitivity.

Description

technical field [0001] The invention relates to a fast and convenient fluorescent PCR detection method for human cytomegalovirus (HCMV), which can simultaneously detect UL122 gene and UL123 gene of HCMV genome, and belongs to the field of molecular biology. Background technique [0002] Human cytomegalovirus (HCMV), also known as human herpesvirus type 5 (Human herpesvirus 5), belongs to the subfamily of a herpesvirus. HCMV is one of the most common pathogens of human congenital virus infection, and can cause central nervous system infection in children. Nervous system involvement, multiple organ damage, mental retardation, deafness, etc.; can cause mononucleosis in normal healthy people; in immunocompromised persons such as organ transplant recipients, acquired immunodeficiency syndrome patients and cancer patients One can cause serious infection, even death. Together with toxoplasma, rubella virus, herpes simplex virus and Treponema pallidum, it is called the five major b...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 何剑军吴大治夏懿
Owner SHANGHAI FOSUN PHARMA (GROUP) CO LTD
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