Kit for genotyping VZV, production method of kit and application of kit

A varicella zoster and kit technology, which is applied in the field of kits for typing and detecting varicella zoster virus, which can solve the problems of difficult differential diagnosis, incapable of high-throughput detection, incapable of batch detection, etc.

Inactive Publication Date: 2015-12-09
CHENGDU MILITARY GENERAL HOSPITAL OF PLA
View PDF4 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Looking back on the previous clinical diagnosis methods of VZV infection, it is found that clinicians mainly rely on typical clinical symptoms and skin lesions to diagnose VZV infection, but other viruses (such as coxsackie virus, herpes simplex virus) infection can also cause chickenpox Pseudomonas rash makes differential diagnosis difficult
However, the existing laboratory diagnostic methods include virus culture, direct fluorescent antibody analysis, PCR, and serum antibody detection, all of which require large sample size, complicated operation, slow speed, incapable of batch detection, multiple detection, low sensitivity, etc. Disadvantages, these are plagued by clinical diagnosis and treatment, prevention
Among them, virus culture is the gold standard for the diagnosis of VZV infection, but it requires special laboratory conditions, complex operation, takes several days, and the positive rate is low; direct fluorescent antibody analysis is cumbersome, requires professional and experienced techn

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kit for genotyping VZV, production method of kit and application of kit
  • Kit for genotyping VZV, production method of kit and application of kit
  • Kit for genotyping VZV, production method of kit and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] The sources and records of the virus strains used for typing in Example 1 are shown in Table 2 below.

[0082] Table 2

[0083]

Embodiment 3

[0084] The sample herpes fluid to be tested used in the clinical verification of Example 3 was collected from patients with chickenpox and herpes zoster admitted to the General Hospital of the Chengdu Military Command of the Chinese People's Liberation Army.

[0085] Embodiment 1, the preparation of kit of the present invention

[0086] 1. Selection of varicella-zoster virus detection and typing sites

[0087] Using the NCBI nucleic acid database ( http: / / www.ncbi.nlm.nih.gov / nuccore / ) search and download the genome sequence of each virus strain of VZV, and type each virus strain according to the Clade typing method. Using DNAman software to conduct multiple sequence comparisons of VZV strains, herpes simplex virus type 1 (HSV1), herpes simplex virus type 2 (HSV2), cytomegalovirus (CMV), Epstein-Barr virus (EBV) strains, and human genome DNA sequences Yes, exclude homologous sequences and find out the specific SNP sites of various VZV strains.

[0088] A total of 21 strai...

Embodiment 2

[0104] Embodiment 2, the verification of kit sensitivity and specificity of the present invention

[0105] Use clade1-5 type VZV standard as the sample to be tested, human genome, herpes simplex virus 1, herpes simplex virus 2, cytomegalovirus, Epstein-Barr virus DNA as the control sample, globin probe as the internal standard, Verify the sensitivity and specificity of the kit of the present invention according to the following steps:

[0106] (1) adopt the specific primer shown in Table 3 of embodiment 1 to carry out multiplex PCR amplification to the genomic DNA of sample to be tested;

[0107] (2) The specific probes shown in Table 3 of Example 1 are respectively coupled with fluorescent microspheres of different numbers to prepare mixed microsphere hybridization solution;

[0108] (3) The hybridization product obtained after mixing the PCR product with the microsphere hybridization solution for hybridization reaction is loaded on the Luminex200 flow cytometry fluorescence...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a kit for genotyping VZV (Varicella-Zoster Viruses). The kit is characterized by comprising a nucleotide sequence shown as the Table 3 in the specification, and specific primers and specific probes corresponding to clade1-5 type VZV of chemical structures. The kit has the function of detecting various kinds of VZV DNA (Deoxyribonucleic Acid); the detection sensitivity is 10<2> copies/reaction; no cross reaction with human genome, herpes simplex viruses type I/type II, cytomegaloviruses and EB viruses exists; and the kit is applicable to the virus gene diagnosis of clinical VZV infected persons, and can also be used for the epidemiology survey of different Clade types of VZV.

Description

technical field [0001] The invention belongs to the technical field of microbial gene diagnosis, and in particular relates to a kit for typing and detecting varicella-zoster virus and its production method and application. Background technique [0002] Varicella-zostervirus (Varicella-zostervirus, VZV) is a virus belonging to the genus Varicellavirus in the family Herpesviridae, and is the common pathogen of clinical varicella and herpes zoster. When a patient is first infected with VZV, chickenpox symptoms will occur, and a lifelong latent infection will occur. After several years, when the body's immunity declines, the latent virus may reactivate to cause herpes zoster. If chickenpox is not diagnosed and treated in time, it will cause secondary varicella pneumonia, encephalitis, myocarditis, etc., and even cause death, while herpes zoster will bring severe pain to patients for a long time, bring a lot of inconvenience to patients' lives, and greatly increase public health ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/686C12Q1/70C12Q1/705C12Q2537/143C12Q2563/149C12Q2563/107
Inventor 吴丽娟刘毓刚
Owner CHENGDU MILITARY GENERAL HOSPITAL OF PLA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap