Detection kit for human herpes viruses EBV and VZV

A detection kit and technology for human herpes are applied in the field of human herpes virus EBV and VZV detection kits, which can solve the problems of low antibody specificity, long time, and low sensitivity.

Inactive Publication Date: 2014-06-18
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional EBV and VZV detection methods mainly include virus isolation and culture and serological detection. The former was once the "gold standard" for the diagnosis of herpes virus, but it takes a long time (3-30 days) and has low sensitivity (especially in the treatment of antiviral drugs). The latter is divided into direct antigen detection method and antibody indirect detection method. The dire

Method used

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  • Detection kit for human herpes viruses EBV and VZV
  • Detection kit for human herpes viruses EBV and VZV

Examples

Experimental program
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Effect test

Embodiment 1

[0028] see figure 1 , the human herpesvirus EBV and VZV detection kit provided by the present invention is composed of quantitative PCR reaction solution 1, EBV standard substance 2, VZV standard substance 3, EBV positive control substance 4, VZV positive control substance 5, negative control substance 6, instructions 7 and box body 8 form.

[0029] The quantitative PCR reaction solution contains PCR buffer, MgCl 2 , dNTPs, thermostable DNA polymerase, EBV upstream amplification primers, EBV downstream amplification primers, VZV upstream amplification primers, VZV downstream amplification primers, EBV fluorescent probes and VZV fluorescent probes.

[0030] The primer sequences for PCR amplification are:

[0031] EBV upstream amplification primer sequence (SEQ ID No: 1): 5'-TCATCTACGGGGACACGGAC-3';

[0032]EBV downstream amplification primer sequence (SEQ ID No: 2): 5'-GAGCTCCACCCCCTTCATC-3';

[0033] VZV upstream amplification primer sequence (SEQ ID No: 3): 5'-TAGGCGGATGG...

Embodiment 2

[0041] Example 2 Sensitivity and specificity test of human herpesvirus EBV and VZV detection kits

[0042] (1) Materials:

[0043] The selected pathogenic microorganisms include: experimental group: EBV (strain B95-8) provided by Beijing Institute of Virology, VZV (strain EF) provided by the microbiology teaching and research group of Anhui Medical University; control group: Staphylococcus aureus, Escherichia coli, type B Hepatitis virus, Cryptococcus neoformans, Candida albicans and human genome were provided by the Bacteria Room, Virus Room and Gene Amplification Room of Children's Hospital Affiliated to Zhejiang University.

[0044] (2) Design and synthesis of primers and probes:

[0045] Conduct bioinformatics analysis on the conserved region sequences of EBV and VZV, design and screen PCR amplification primers and specific fluorescent probes, and entrust Shanghai Sangon Biotech Co., Ltd. to synthesize them.

[0046] EBV upstream amplification primer sequence (SEQ ID...

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Abstract

The invention provides a detection kit for human herpes viruses EBV and VZV. The detection kit comprises quantitative PCR reaction solution, an EBV standard, a VZV standard, an EBV positive control, a VZV positive control, a negative control, instructions and the kit body, the quantitative PCR reaction solution comprises PCR buffer, MgCl2, dNTPs, heat-resistant DNA polymerase, an EBV forward amplification primer, an EBV reverse amplification primer, a VZV forward amplification primer, a VZV reverse amplification primer, a EBV fluorescent probe and a VZV fluorescence probe. The human herpes viruses EBV and VZV can be detected in one step by using real-time quantitative PCR and two-color fluorescent probes according to the detection kit, synchronous diagnosis of EBV and VZV infections is realized, positive virus can be accurately quantified in real time, urgent need for accurate diagnosis of EBV and VZV infections in the early clinical diagnosis is met and a basis is provided for timely definitive therapy of EBV and VZV infections.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a fluorescent quantitative PCR detection kit, in particular to a dual-probe real-time fluorescent quantitative PCR for simultaneously detecting Epstein-Barr virus (Epstein-Barr virus, EBV) and varicella-zoster virus (varicella-zoster virus, VZV) diagnostic kits. Background technique [0002] EBV and VZV are common human herpesviruses that cause viral encephalitis in children. After EBV invades the central nervous system, it can lead to cerebral edema, hemorrhage, deposition of immune complexes, and demyelination-like changes in brain tissue. The clinical manifestations can be acute onset or chronic active damage. Acute EBV encephalitis is mostly caused by the virus directly invading the nervous system, such as meninges, brain tissue, spinal cord, and nerve axons in multiple parts of peripheral nerves; chronic EBV encephalitis is related to autoimmune reactions, which is the de...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/70C12Q1/686C12Q2531/113C12Q2561/113C12Q2563/107
Inventor 陶然尚世强舒强杜立中李伟
Owner ZHEJIANG UNIV
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