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Quick high-flux dengue fever virus detection typing method

A dengue virus and typing method technology, applied in the field of in vitro diagnostic reagents, can solve the problems of increasing false positives and complicated operations, and achieve the effects of improving detection capabilities, rapid and accurate detection, and reducing time and complexity

Active Publication Date: 2015-04-01
SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The current gene chip detection for dengue fever virus detection has been used in some researches, but the probes of these gene chips mostly use manual mode, which is complicated to operate and increases the chance of many false positives

Method used

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  • Quick high-flux dengue fever virus detection typing method
  • Quick high-flux dengue fever virus detection typing method
  • Quick high-flux dengue fever virus detection typing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Detection specificity and sensitivity of four serotypes of dengue virus

[0075] 1. Experimental steps

[0076] 1. Collection of blood samples

[0077] Take 1.0ml of whole blood sample from the patient for port testing, and centrifuge to obtain serum.

[0078] 2. Preliminary Identification of Blood Samples

[0079] The colloidal gold method was used to preliminarily determine whether the patient was infected with dengue fever.

[0080] 3. Genomic RNA Extraction

[0081] Dengue RNA was extracted using a blood sample genomic RNA extraction kit (spin column type, Qiagen). Take 100uL serum sample, and extract RNA according to the extraction steps in the kit instruction manual.

[0082] 4. Determination of RNA Concentration

[0083] RNA concentration was quantified by real-time quantitative PCR using ABI's real-time fluorescence detector.

[0084] 5. Amplification of target genes

[0085] 5.1 Design of primers and probes

[0086] Primers were designed for ...

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Abstract

The invention relates to a quick high-flux dengue fever virus detection typing method. The detection method integrates the two powerful molecular biology techniques of polymerase chain reaction (PCR) and microarray, and directly fixes the PCR hybridized probe to the hybridization bin of the microarray in the same chip as the PCR reaction chamber. The detection method comprises the following steps: specimen RNA (ribonucleic acid) extraction, PCR amplification, hybridization, cleaning and result interpretation. The method can perform quick and sensitive detection and typing on dengue fever virus (serum type I, II, III and IV). The method can greatly enhance the detection efficiency of the front-line inspection health officers of import-export ports, reduce the workload and maximally solve the problem of possible undetected positive in the traditional detection method, thereby maximally preventing the dengue epidemic situation from generation.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnostic reagents, in particular to a rapid and high-throughput dengue virus detection and typing method. Background technique [0002] Dengue virus (DV) is a member of the Flaviviridae family (Flav iv iridae), along with yellow fever virus (YEV), Japanese encephalitis virus (JEV) and hepatitis C virus (HCV), all of which can cause severe human disease. cause major public health problems. Dengue virus infection mainly occurs in tropical and subtropical regions, and can cause symptoms such as dengue fever (DF) and dengue hemorrhagic fever / dengue shock syndrome (DHF / DSS). About 50 million to 100 million people are infected and 25,000 people die every year. . Since the 1980s, the epidemic has become more and more serious. There are more than 100 countries in the world. The epidemic is most serious in Southeast Asia and the Western Pacific region. There are more than one million cases in the worl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6837C12Q1/70C12Q2531/113C12Q2565/501Y02A50/30
Inventor 田桢干张子龙李深伟王传现李平王俐赵百慧
Owner SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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