Hand-foot-and-mouth disease (HFMD) related virus parallel detection liquid-phase chip and preparation method and application thereof

A parallel detection and liquid phase chip technology, which is applied in the field of molecular biology technology and clinical detection, can solve the problems of inconvenience, tedious and time-consuming laboratory and clinical diagnosis, and achieve the effect of saving detection cost

Inactive Publication Date: 2010-01-20
SHANDONG CENT FOR DISEASE CONTROL & PREVENTION +1
View PDF0 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] According to the characteristics of diversification of viruses related to hand, foot and mouth disease, the current ELISA, RT-PCR and real-time RT-PCR methods used in the detection of hand, foot and mouth disease can only detect one virus at a time. And the needs of clinical diagnosis, the problem to be solved in the present invention is to provide a kind of hand, fo

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0026] Example 1: Coupling of probes with microspheres with known numbers

[0027] 1. Select 26, 36, and 46 carboxyl microspheres (Luminex Company) respectively, and use a vortex shaker to shake the microsphere suspension for 20s to make the microspheres evenly mixed.

[0028] 2. Take the above carboxyl microspheres about 2×10 respectively 3 Each was transferred to a centrifuge tube, and centrifuged at ≥8000×g for 2 minutes to precipitate carboxyl microspheres.

[0029] 3. Remove the supernatant and add 100μl dH 2 O, resuspend the microspheres with a vortex shaker for 20 seconds, centrifuge for 2 minutes at ≥8000×g, and precipitate the carboxyl microspheres.

[0030] 4. Remove the supernatant, add 80 μl, 100 mM, pH=6.2 sodium dihydrogen phosphate solution, and resuspend the washed carboxyl microspheres with a vortex shaker for 20 seconds.

[0031] 5. Add 10μl, 50mg / ml Sulfo-NHS (using dH 2 O dilution), gently shake with a vortex shaker.

[0032] 6. Add 10μl, 50mg / ml EDC (using dH 2 O dil...

Example Embodiment

[0043] Example 2: Application of the liquid-phase chip for parallel detection of hand, foot and mouth disease-related viruses of the present invention in clinical detection

[0044] The technical process of using the hand-foot-mouth disease-related virus parallel detection liquid-phase chip to detect hand-foot-mouth disease-related virus according to the present invention:

[0045] 1. Extraction of viral nucleic acid: RNA can be extracted using commercial kits, such as QIGEN ViralRNA mini Extraction Kit (CAT: 52904) to extract viral RNA from clinical specimens or virus isolation cultures;

[0046] (1) Add 560μl of AVL buffer to a 1.5ml centrifuge tube;

[0047] (2) Add 140μl of clinical specimen or virus isolation culture to this centrifuge tube;

[0048] (3) Place it at room temperature for 10 minutes, and then centrifuge immediately;

[0049] (4) Add 560μl of absolute ethanol, mix well for at least 15s, and centrifuge immediately;

[0050] (5) Carefully add about 630 μl of the mixed sol...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a hand-foot-and-mouth disease (HFMD) related virus parallel detection liquid-phase chip and a preparation method and an application thereof. The HFMD related virus parallel detection liquid-phase chip in the invention mainly comprises microballoons, specific probes, biotinylated universal primer and streptavidin-phycoerythrin, which is characterized in that microballoons in the chip are Number 26, Number 36 and Number 46 microballoons; the specific probes are aminated specific probes aiming at virus VP1 area of EV71, CoxA16 and Echo9; the universal primer is enterovirus universal primer marked by activated biotin. In the invention, different probes can combine with different target molecules, different detection reactions can be classified by using laser for detecting color numbers of sphere matrix after reaction, thus having the quantitative or qualitative characteristics and high throughput advantage, thereby saving time and power and detection cost greatly.

Description

(1) Technical field [0001] The invention relates to a liquid phase chip and its preparation method and application, in particular to a liquid phase chip for parallel detection of hand-foot-mouth disease-related viruses and its preparation method and application, belonging to the technical fields of molecular biology technology and clinical detection. (2) Background technology [0002] Hand-foot-mouth disease (HFMD), also known as exanthematous stomatitis, is caused by enterovirus infection, and its clinical manifestations are mainly characterized by fever and rashes or herpes on the hands, feet, and oral cavity. An acute common infectious disease. It is more common in children, and infants and young children under 3 years old are more likely to get sick. HFMD is highly contagious, with complex transmission routes and rapid transmission, which can cause a pandemic in a short period of time. [0003] The pathogens causing HFMD are mainly types 2, 4, 5, 7, 9, 10, and 16 of th...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64C12R1/93
Inventor 李忠张华宁吕慧王显军毕振强张国宁张凯宁汪朝晖李艳艳
Owner SHANDONG CENT FOR DISEASE CONTROL & PREVENTION
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap