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Method for detecting dengue by using gene chip method

A technology of gene chip and dengue fever is applied in the field of virus inspection to achieve the effect of improving efficiency, saving time and avoiding repeated cultivation

Inactive Publication Date: 2012-12-05
中华人民共和国天津出入境检验检疫局
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no effective treatment for dengue fever, so rapid detection and timely control of the source of infection are of great significance for effective control of the disease

Method used

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  • Method for detecting dengue by using gene chip method
  • Method for detecting dengue by using gene chip method
  • Method for detecting dengue by using gene chip method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Sample: A sample to be tested.

[0054] 1. Sample Processing

[0055] Mix the sample for processing.

[0056] 2. Nucleic acid extraction

[0057] Take the sample and resuspend it in PBS, and use the RNA extraction kit to perform RNA extraction according to the instructions.

[0058] 3. Preparation of cDNA

[0059] The total RNA was reverse-transcribed into cDNA using an RNA reverse transcription kit according to the instructions.

[0060] 4.PCR amplification

[0061] The composition ratio of each component in the reaction system in the isothermal amplification system is as follows

[0062] The composition ratio of each component in the nucleic acid amplification reaction system is as follows:

[0063]

[0064] The procedure for nucleic acid amplification is:

[0065] (1) 95°C 10min

[0066] (2) 95°C 30s

[0067] (3) 55°C 20s

[0068] (4) 72°C 30s

[0069] (5) Go back to step (2) and repeat 35 times

[0070] (6) 72°C 2min

[0071] 5. Prehybridization

[00...

Embodiment 2

[0082] Sample: A sample to be tested.

[0083] 1. Sample Processing

[0084] Mix the sample for processing.

[0085] 2. Nucleic acid extraction

[0086] Take the sample and resuspend it in PBS, and use the RNA extraction kit to perform RNA extraction according to the instructions.

[0087] 3. Preparation of cDNA

[0088] The total RNA was reverse-transcribed into cDNA using an RNA reverse transcription kit according to the instructions.

[0089] 4.PCR amplification

[0090] The composition ratio of each component in the reaction system in the isothermal amplification system is as follows

[0091] The composition ratio of each component in the nucleic acid amplification reaction system is as follows:

[0092]

[0093]

[0094] The procedure for nucleic acid amplification is:

[0095] (1) 95°C 10min

[0096] (2) 95°C 30s

[0097] (3) 55°C 20s

[0098] (4) 72°C 30s

[0099] (5) Go back to step (2) and repeat 35 times

[0100] (6) 72°C 2min

[0101] 5. Prehybrid...

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PUM

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Abstract

The invention discloses a method for detecting dengue by using a gene chip method. According to the method, sequences of universal primers are designed, namely 5'ACAGCMGGIITGGGACACAAGAAT and 5'GGTTTRACRCARTCRTCYCCRCT; and specific probes for detecting four dengue virus are designed, namely 5'CCAACCTAGCTGAGAGYGRTCTTGACT, 5'TGGTAACYAACCACATGYAAGGATAACARAAGA, 5'AGGYGTGTTYTCYAAGYCAGACCTCGAGAACCC and 5'AAGGGCTGAAAGAAAGAGTTGAGAAATGGCTGA. Aiming at the dengue, the defects in the prior art are overcome, and a method for detecting the dengue quickly in high flux and in a typing mode is provided.

Description

technical field [0001] The invention relates to virus inspection technology, in particular to a method for detecting dengue fever virus by using a gene chip method. Background technique [0002] Dengue fever is an acute infectious disease caused by dengue virus and transmitted by Aedes mosquitoes. The clinical features are sudden onset, high fever, muscle, bone marrow and joint pain all over the body, extreme fatigue, and some patients may have rash, bleeding tendency and enlarged lymph nodes. The disease was discovered in Cairo, Egypt, Jakarta, Indonesia, and Philadelphia, USA in 1779, and was named joint fever and fracture fever according to the symptoms. Dengue fever was named by the Royal Society of Physicians in London in 1869. In 1943, Japanese scientists discovered dengue virus for the first time, and the United States also discovered the virus one after another. Its etiology was not understood until 1944. Dengue virus was isolated for the first time in 1952, and d...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCY02A50/30
Inventor 杨春江祁军柴洪森詹曦菁刘寅窦勇鹰周君野
Owner 中华人民共和国天津出入境检验检疫局