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Kit for detecting Zika virus, dengue virus and chikungunya virus

A kit, primer and probe technology, applied in the field of kits for detection of Zika virus, dengue fever virus and chikungunya virus, can solve the problems of reagent cold chain transportation difficulties and application limitations

Active Publication Date: 2017-05-31
CAPITALBIO CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the frequent occurrence of mosquito-related viruses, such as Zika virus, dengue virus, and chikungunya virus, etc., are concentrated in tropical areas such as Africa, South America, and Southeast Asia, and are limited by local climatic conditions and economic conditions, most places require It is very difficult to realize the cold chain transportation of reagents, which greatly restricts the application of these nucleic acid detection reagents in these regions

Method used

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  • Kit for detecting Zika virus, dengue virus and chikungunya virus
  • Kit for detecting Zika virus, dengue virus and chikungunya virus
  • Kit for detecting Zika virus, dengue virus and chikungunya virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0119] Embodiment 1, the preparation of kit

[0120] 1. Preparation of primers and probes

[0121] The primer probe set A for detecting Zika virus (Zika Virus) is composed of primer ZIKV-F, primer ZIKV-R and probe ZIKV-P. Primer ZIKV-F is shown as sequence 1 in the sequence listing. The primer ZIKV-R is shown as sequence 2 in the sequence listing. The nucleotide sequence of the probe ZIKV-P is shown as sequence 3 in the sequence listing, the 5' end has a fluorescent group JOE, and the 3' end has a quencher group BHQ2.

[0122] ZIKV-F: 5'-CAGCTGGCATCATGAAGAAYC-3';

[0123] ZIKV-R: 5'-CACYTGTCCCATCTTYTTCTCC-3';

[0124] ZIKV-P: 5'-JOE-CYGTTGTGGATGGAATAGTGG-BHQ2-3'.

[0125] Primer probe set B for detecting dengue virus (Dengue virus) consists of primer DENV-F, primer DENV-R1, primer DENV-R2 and probe DENV-P. The primer DENV-F is shown as sequence 4 in the sequence listing. The primer DENV-R1 is shown as sequence 5 in the sequence listing. The primer DENV-R2 is shown as s...

Embodiment 2

[0151] Example 2. Sensitivity and specificity analysis for three kinds of mosquito-borne virus nucleic acid detection kits

[0152] 1. Template preparation

[0153] A Zika virus RNA standard product (single-stranded RNA molecule, whose nucleic acid sequence is shown in sequence 11 in the sequence listing) was prepared as RNA1.

[0154] A dengue virus RNA standard product (single-stranded RNA molecule, see sequence 12 in the sequence listing for its nucleic acid sequence) was prepared as RNA2-1. RNA2-1 corresponds to dengue virus types Ⅰ, Ⅱ and Ⅲ.

[0155] A dengue virus RNA standard product (single-stranded RNA molecule, whose nucleic acid sequence is shown in sequence number 14 in the sequence listing) was prepared as RNA2-2. RNA2-2 corresponds to dengue virus type Ⅳ.

[0156] Prepare a chikungunya virus RNA standard product (single-stranded RNA molecule, whose nucleic acid sequence is shown in sequence number 13 in the sequence listing) as RNA3.

[0157] Get RNA (RNA1 or...

Embodiment 3

[0170] Embodiment 3, the specificity analysis of kit

[0171] Common pathogens with similar symptoms to the three mosquito-borne viral infections were used as specificity references.

[0172] The samples to be tested are as follows:

[0173] Hepatitis B virus: Zhang Juan, Chen Jianzong, Zhang Jinping, Jia Xin, Jiang Weifeng; Anti-hepatitis B virus effect of astragaloside IV in vitro; Fourth Military Medical University, 2008,28(24):2291-2293.

[0174] Epstein-Barr virus (human herpesvirus type 4): Wu Fan, Zhai Weiwei, Ge Liuying, Qi Yanwei, Gao Hui; Analysis of the detection of human herpesvirus types 1-4 in the saliva of 245 cases of human immunodeficiency virus infection; "West China Journal of Stomatology" , 2012, 30(5):514-517.

[0175] Measles virus L4 strain: Zhao Dapeng, Yao Weimin, Wu Xiaojuan, Qi Fengchun, Wang Chunyi; Determination of the complete genome sequence of measles virus L4 strain; National Conference on Biological Products, 2007.

[0176] H5N1 subtype avi...

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Abstract

The invention discloses a kit for detecting Zika virus, dengue virus and chikungunya virus. The invention discloses a primer and probe combination, and the primer and probe combination includes a primer and probe group I, a primer and probe group II and a primer and probe group III; the primer and probe group I is composed of a primer ZIKV-F, a primer ZIKV-R and a probe ZIKV-P; the primer and probe group II is composed of a primer DENV-F, a primer DENV-R1, a primer DENV-R2 and a probe DENV-P; the primer and probe group III is composed of a primer CHIKV-F, a primer CHIKV-R and a probe CHIKV-P; and the primer ZIKV-F, the primer ZIKV-R, the probe ZIKV-P, the primer DENV-F, the primer DENV-R1, the primer DENV-R2, the probe DENV-P, the primer CHIKV-F, the primer CHIKV-R and the probe CHIKV-P are sequentially shown as sequences 1-10. The kit provided by the invention has a great value for detection of the Zika virus, the dengue virus and the chikungunya virus and for prevention and control of related diseases.

Description

technical field [0001] The invention relates to a kit for detecting Zika virus, dengue fever virus and chikungunya virus. Background technique [0002] Mosquito-borne viruses are widely distributed around the world and are important pathogens of infectious diseases. They are mainly transmitted through mosquito bites, and the transmission speed is fast, which can easily cause serious harm to humans and society. Especially in recent years, mosquito-borne virus outbreaks have continued. Most of the mosquito-borne viruses belong to the genus Yellow fever virus of the family Yellow fever virus, mainly including yellow fever virus, dengue virus and Zika virus, and a few belong to the genus Alphavirus of the family Hymenaviridae, such as Chikungunya virus. The virus is transmitted by the media, and in recent years, there have been outbreaks of epidemics. From 2015 to 2016, the Zika virus outbreak broke out in Brazil and quickly spread to more than 30 countries and regions. At the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/701C12Q2521/107C12Q2531/113C12Q2545/113C12Q2563/107Y02A50/30
Inventor 张岩盖伟单万水邢婉丽宋翠丹马桂红刘厚明程京
Owner CAPITALBIO CORP
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