Human herpes virus six-type real-time fluorescence quantitative PCR (polymerase chain reaction) kit

A human herpes virus, real-time fluorescence quantitative technology, applied in the determination/inspection of microorganisms, microorganisms, microorganism-based methods, etc., can solve the problem of inability to detect virus infection, and achieve a short detection period, high efficiency, and strong specificity. Effect

Inactive Publication Date: 2011-08-17
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Qualitative PCR, similar to IgG antibody titration, cannot detect latent virus infection in most healthy people when tested on peripheral blood leukocytes, and only quantitative PCR may be applicable to the detection of human herpesvirus 6 virus DNA in body fluids

Method used

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  • Human herpes virus six-type real-time fluorescence quantitative PCR (polymerase chain reaction) kit
  • Human herpes virus six-type real-time fluorescence quantitative PCR (polymerase chain reaction) kit
  • Human herpes virus six-type real-time fluorescence quantitative PCR (polymerase chain reaction) kit

Examples

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Embodiment Construction

[0067] Below in conjunction with accompanying drawing and embodiment describe in detail:

[0068] 1. Example 1---Human herpesvirus type 6 real-time fluorescent quantitative PCR kit

[0069] 1) The composition of the kit:

[0070] A pair of specific primers F and R, a specific fluorescent probe FP, HSV type positive control, RNase FreeH 2 O negative control, 5xPCR Buffer, Enzyme Mix

[0071] 2) The primers designed for the specific amplification gene sequence of human herpesvirus type 6 are:

[0072] The specific primers designed according to the U67 open reading frame of HHV6 are:

[0073] F: 5'-ACAAAGCGAAATTATTCCAGAGCGT-3' 24bp

[0074] R: 5'-GCGCTAGGTTGAGGATGATCGC-3' 22bp

[0075] The total length of the amplified gene sequence is 133bp;

[0076] The specific probes designed according to the U67 open reading frame of HHV6 are:

[0077] FP: 5'FAM-ACACCAGACGTCACACCCGAAGGAATT-TAMARA 3'27bp

[0078] 3) 5xPCR Buffer preparation:

[0079]

[0080]

[0081] Store in ...

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Abstract

The invention provides a human herpes virus six-type real-time fluorescence quantitative PCR (polymerase chain reaction) kit, and relates to a PCR kit in the technical field of the biology. The kit comprises a pair of specific primers, a specific fluorescent probe, a positive control, a negative control, a 5*PCR Buffer and an Enzyme Mix, wherein the specific primers are designed to be: F: 5'-ACAAAGCGAAATTATCCAGAGCGT-3', and R: 5'-GCGCTAGGTTGAGGATGATCGC-3'; and the specific probe is designed to be: FP: 5'-FAM-ACACCAGACGTCACACCCGAAGGAATT-TAMARA3'. The kit is short in detection period, high in efficiency, high in viral specificity detection, high in accuracy, qualitative and quantitative in virus analysis, higher in the sensitivity compared with the common PCR and immunology detection method, simple in operation, easy to popularize, and good in the repeatability of the experimental result.

Description

technical field [0001] The invention relates to a PCR ((Polymerase Chain Reaction, polymerase chain reaction) kit in the field of biotechnology, in particular to a human herpesvirus type 6 real-time fluorescent quantitative PCR kit. Background technique [0002] Human herpesvirus type 6 (Human Herpers Viruses, HHV-6), belonging to the Herpesviridae β subfamily, is a linear double-stranded DNA virus. Human herpesvirus 6 is a ubiquitous pathogen that was isolated in 1986 from a patient with lymphoproliferative disease. Serological epidemiological surveys show that human herpesvirus 6 is highly prevalent in the population, and the main infected population is infants and young children aged 6 months to 2 years. Viral infection causes fever and rash in infants and young children, and occasionally meningitis or chronic hepatitis. [0003] In immunocompetent adults, human herpesvirus 6 etiology has been proposed for several pathological entities, such as encephalitis, fulminant h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12R1/93C12Q1/68
Inventor 吴建国刘为勇艾洪武石康刘映乐汤行春熊鹰项荣程议锋李彤亚章琪程念张家云何茜妮
Owner WUHAN UNIV
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