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Kit for flavivirus quick typing and virus load detection

A viral load and flavivirus technology, which is applied in the determination/inspection of microorganisms, resistance to vector-borne diseases, DNA/RNA fragments, etc., can solve the problems that cannot meet the detection requirements of flaviviruses, and facilitate prevention, control and treatment Measures, reducing sample size requirements, and improving the effect of cure rate

Inactive Publication Date: 2016-11-09
GUANGZHOU EIGHTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the detection kit can detect three flaviviruses at the same time, it still cannot meet the clinical detection needs of flaviviruses

Method used

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  • Kit for flavivirus quick typing and virus load detection
  • Kit for flavivirus quick typing and virus load detection
  • Kit for flavivirus quick typing and virus load detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, primers and probes

[0039] The invention provides a primer and a probe for rapid typing of flavivirus and detection of viral load.

[0040] Table 1 Primers and probes provided by the present invention

[0041]

Embodiment 2

[0042] Embodiment 2, the detection method that is used for the test kit of flavivirus rapid typing and viral load detection

[0043] 1. Prepare a kit including the following components: 1 tube of reaction solution I (20 μl / tube), detection reaction II (280 μl / tube), 1 tube of positive quality control (20 μl / tube), and 1 tube of negative quality control (20 μl / tube). tube) 1 tube, quantitative reference substances S1 ~ S4 (20μl / tube) 1 tube each.

[0044] 2. Specimen type: Serum, urine and saliva samples collected clinically.

[0045] 3. Nucleic acid extraction:

[0046] Commercial RNA extraction kits can be used, such as nucleic acid extraction reagents based on silica gel membrane spin column method or nucleic acid extraction reagents based on magnetic bead method, and operate according to the kit instructions, and finally collect 20-30 μl RNA solution for direct detection Or store at -20°C.

[0047] 4. Real-time fluorescence rapid PCR and quantitative detection:

[0048]...

Embodiment 3

[0054] Embodiment 3, the detection clinical sample of using the kit of rapid identification of flavivirus and detection of viral load

[0055] 1. Select 1 case of serum and urine samples from positive cases of Zika virus infection confirmed by clinical and virus culture methods, and 1 case of saliva samples from Zika virus infection positive cases on the 2nd, 3rd, 4th, and 5th day of onset. Serum samples from positive cases identified as chikungunya virus, dengue virus type I, dengue virus type II, dengue virus type III, dengue virus type IV, and yellow fever virus were identified by clinical and virus culture methods. , as well as one case of serum samples from positive cases of measles virus, Hantaan virus, Seoul virus, fever with thrombocytopenia syndrome bunya virus infection, and one case of yellow fever virus vaccine strain. Nucleic acid extraction was performed on all samples, and the steps of on-board amplification detection and result analysis were carried out with re...

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Abstract

The invention belongs to the field of microbial molecular detection, and particularly relates to a kit for flavivirus quick typing and virus load detection. The kit for flavivirus quick typing and virus load detection comprises specific primers and probes for flaviviruses, including specific primers and a probe for dengue virus, specific primers and a probe for Zika virus, specific primers and a probe for yellow fever virus, and specific primers and a probe for Chikungunya virus. The kit for flavivirus quick typing and virus load detection has the advantages of high detection speed and accurate detection and quantification results, can simultaneously detect 3 different flavivirus pathogens and 1 togavirus pathogen capable of causing similar clinical symptoms at one time, can detect and diagnose flavivirus-pathogen-infected suspicious cases in time, and enhances the detection accuracy of flavivirus pathogen infection.

Description

technical field [0001] The invention belongs to the field of microbial molecular detection, in particular to a kit for rapid typing of flaviviruses and detection of viral load. Background technique [0002] Flaviviruses are widely distributed all over the world. They mainly transmit viruses through mosquito bites. They have the characteristics of fast transmission and serious illness, and cause serious harm to human beings and society. With the development of national trade and tourism, the inbound and outbound vehicles and goods surge, and the potential danger of mosquito-borne diseases is gradually increasing. There are many types of flaviviruses, how to detect and identify the types of flaviviruses in the shortest time is of great significance for clinical diagnosis and entry-exit inspection and quarantine. [0003] Traditional diagnostic methods for flaviviruses are mainly detected through serological tests, nucleic acid detection, virus culture or immunological detecti...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/686C12Q1/701C12Q2600/16C12Q2537/143C12Q2563/107C12Q2545/113Y02A50/30
Inventor 张复春高秀洁赵令斋
Owner GUANGZHOU EIGHTH PEOPLES HOSPITAL
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