Method for detection and typing of dengue virus, special chip and kit

A dengue virus and kit technology, applied in biochemical equipment and methods, microbial determination/inspection, resistance to vector-borne diseases, etc., can solve the problems of long time and complicated operation, and achieve low cost and good specificity. Effect

Inactive Publication Date: 2011-12-21
SHENZHEN ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, gold nanoparticle-based detection methods usually require an additional silver staining step, which is cumbersome and time-consuming

Method used

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  • Method for detection and typing of dengue virus, special chip and kit
  • Method for detection and typing of dengue virus, special chip and kit
  • Method for detection and typing of dengue virus, special chip and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1, the structure of biochip and its preparation

[0064] 1. The structure of the biochip

[0065] 1. The biochip is composed of a biochip base and detection probes connected thereto, positive quality control probes (PC), negative quality control probes (NC) and magnetic labels;

[0066] 2. The detection probes are shown in 1) and 2) below:

[0067] 1) a universal probe for detecting dengue virus, the core hybridization sequence of which is shown in SEQ ID NO: 5;

[0068] 2) There are four types as follows:

[0069] 2)-1. The probe for detecting type I dengue virus, its core hybridization sequence is shown in SEQ ID NO: 1,

[0070] 2)-2. The probe for detecting type II dengue virus, its core hybridization sequence is shown in SEQ ID NO: 2,

[0071] 2)-3. The probe used to detect type III dengue virus, its core hybridization sequence is shown in SEQ ID NO: 3,

[0072] 2)-4. The probe for detecting type IV dengue virus, the core hybridization sequence of wh...

Embodiment 2

[0116] Embodiment 2, the kit for detecting dengue virus serotype

[0117] 1. The composition of the kit is as follows:

[0118] 1. The biochip in Example 1.

[0119] 2. The primer pair composition of the target band in the amplified sample, as shown in I to V:

[0120] I. A pair of primers shown in SEQ ID NO: 8 and SEQ ID NO: 9; the amplified product of the primer pair I type dengue virus can hybridize with the probe DEN-1.

[0121] II. A pair of primers shown in SEQ ID NO: 10 and SEQ ID NO: 11; the amplified product of the primer pair of type II dengue virus can hybridize with the probe DEN-2.

[0122] III. A primer pair shown in SEQ ID NO: 12 and SEQ ID NO: 13; the amplified product of the primer pair III dengue virus can hybridize with the probe DEN-3.

[0123] IV. A primer pair shown in SEQ ID NO: 14 and SEQ ID NO: 15; the amplification product of the primer pair IV dengue virus can hybridize with the probe DEN-4.

[0124] V, a pair of primers shown in SEQ ID NO: 6 and...

Embodiment 3

[0136] Embodiment 3, the application of kit

[0137]Type I dengue virus positive samples: Serum samples from people infected with type I dengue virus but not infected with other types of dengue virus, taken from the Shenzhen Center for Disease Control and Prevention, all with the consent of the patient; identification methods see: KRGurukumar, D Priyadarshini, JA Patil, A Bhagat, A Singh. Development of real time PCR for detection and quantitation of Dengue Viruses. Virology Journal, 2009, 1, 23: 1-8).

[0138] Type II dengue virus positive samples: Serum samples from people infected with type II dengue virus but not infected with other types of viruses, obtained from the Guangdong Provincial Center for Disease Control and Prevention, with the consent of the patients; for identification methods see: KR Gurukumar, D Priyadarshini, JA Patil, A Bhagat, A Singh. Development of real time PCR for detection and quantitation of Dengue Viruses. Virology Journal, 2009, 1, 23: 1-8).

[...

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Abstract

The invention discloses a dengue virus detection and parting method, a special chip and a kit. The kit comprises a biological chip and target segment primer pair compositions: primer pairs SEQ ID NO:6 and SEQ ID NO:7; primer pairs SEQ ID NO:8 and SEQ ID NO:9; primer pairs SEQ ID NO:10 and SEQ ID NO:11; primer pairs SEQ ID NO:12 and SEQ ID NO:13; and primer pairs SEQ ID NO:14 and SEQ ID NO:15, wherein the target segment primer pair compositions are hybridized with probes in amplification samples. The kit can detect dengue viruses and can carry out parting identification on the dengue viruses, the dengue viruses can be classified and directly identified into dengue viruses, first dengue viruses, second dengue viruses, third dengue viruses and fourth dengue viruses, the dengue virus classification experiment is carried out on the basis of the method, a better effect is obtained, the true positive rate reaches 100 percent, and the true negative rate reaches 100 percent. The accuracy rate of the four-type dengue virus identification reaches 100 percent.

Description

technical field [0001] The invention relates to a method for detecting and typing dengue virus, a special chip and a kit. Background technique [0002] Dengue virus (dengue virus) belongs to a serotype subgroup of the Flavivirus genus in the Flaviviridae family. Its shape and structure are similar to JE virus, but its size is smaller, about 17-25nm. The dengue virus genome RNA contains about 11,000 nucleotides, and its 5' end is a type I cap structure, and the 3' end lacks a poly(A) tail. Both the 5' end and the 3' end of the genome have a non-coding region. The genome has only one open reading frame, and its 5' end 1 / 4 encodes three structural proteins (C, PrM, and E) of the virus, and the 3' end 3 / 4 encodes seven nonstructural proteins (NS1, NS2a, NS2b, NS3, NS4a, NS4b and NS5). Dengue virus is divided into 1 to 4 serotypes (I, II, III, IV) according to the antigenicity of the viral envelope protein E. There is cross-antigenicity among various types of viruses, but no c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCY02A50/30
Inventor 史蕾顾大勇徐云庆刘春晓赵纯中杨燕秋冬冰
Owner SHENZHEN ACAD OF INSPECTION & QUARANTINE
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