Test paper strip technology for detecting specific DNA (Deoxyribonucleic Acid) by applying SYBR Green I fluorescent dye

A fluorescent dye and specific technology, applied in the field of biotechnology applications, can solve the problems of complex labeling steps, inability to perform multiple detection, etc., and achieve the effects of reduced detection cost, rapid result display, and strong specificity.

Active Publication Date: 2014-07-02
艾吉泰康(嘉兴)生物科技有限公司
View PDF6 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the labeling steps of this method are com

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

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Test paper strip technology for detecting specific DNA (Deoxyribonucleic Acid) by applying SYBR Green I fluorescent dye
  • Test paper strip technology for detecting specific DNA (Deoxyribonucleic Acid) by applying SYBR Green I fluorescent dye
  • Test paper strip technology for detecting specific DNA (Deoxyribonucleic Acid) by applying SYBR Green I fluorescent dye

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] This example aims to detect influenza viruses A, B, H1, H3 and their mixtures. The nucleotide sequences of specific probes and target DNA are shown in Table 1.

[0032] Table 1. Influenza virus A, B, H1, H3 probes and target DNA sequences

[0033]

[0034] The detection method of this embodiment comprises the following steps:

[0035] (1) Selection of test strip material

[0036] Select nylon membrane (ZetaProbe TM , 15cm×15cm sheet, Bio-Rad) As the material of the test strip, the nylon membrane was cut into a strip of 3cm×0.5cm.

[0037] (2) Preparation of probe sensing layer

[0038] Spot the probes A, B, H1, H3 (500nM, 0.5μL) on the cut nylon membrane at fixed intervals, and then irradiate the nylon membrane with 254nm ultraviolet light (Unitec LF-206LS) for 2 minutes, 60 Bake at ℃ for 5 minutes, and finally store in a dark and dry place.

[0039] (3) The target sequence of the sample to be tested is obtained

[0040] Primers were designed using probe A as ...

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 provides a technology for rapidly detecting specific DNA (Deoxyribonucleic Acid) by applying an SYBR Green I fluorescent dye to a test paper strip. The technology comprises the following steps of selecting a nylon membrane as a material of the test paper strip; pointing probes onto the nylon membrane, then, irradiating with ultraviolet rays, baking, drying in the dark and storing, wherein the probes are specific nucleotide sequences to be detected; carrying out PCR (Polymerase Chain Reaction) amplification in a manner of taking samples to be detected as templates and taking primers, which are designed by taking the probes as templates, as primers, then, carrying out denaturation for 10 minutes at the temperature of 94 DEG C, and immediately lowering by 4 DEG C, so as to obtain sequences to be detected; flushing the nylon membrane with a buffer solution, dispensing a hybridization solution to the nylon membrane, hybridizing in the dark at room temperature, cleaning the nylon membrane sequentially with a buffer solution and deionized water, imaging with a 488nm-laser imaging scanner, and judging a result in accordance with whether the nylon membrane emits light or not, wherein the hybridization solution contains SYBR Green I and the sequences to be detected. The technology is simple, inexpensive and rapid and is applicable to clinical rapid diagnosis.

Description

technical field [0001] The invention relates to the application field of biotechnology, in particular to a technology for rapid detection of specific DNA on a test strip by using SYBR Green I fluorescent dye. Background technique [0002] Pathogen detection has always been of great value in many research fields, especially in the medical and food industries. Rapid and accurate detection and identification of pathogens is the key to effective infection control and early treatment. However, standard pathogen culture requires not only incubation on medium for at least 8 hours, but also additional biochemical or immune recognition, which is tedious and time-consuming. In contrast, nucleic acid-based assays such as quantitative polymerase chain reaction (qPCR) and microarrays are faster and more sensitive, however, many of these methods are still complex and expensive. [0003] In recent years, reported lateral flow assays have contributed greatly to the development of paper-ba...

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): C12Q1/68
CPCC12Q1/6837C12Q2563/107C12Q2565/501C12Q2565/625
Inventor 李孟皇边红武韩凝武金霞
Owner 艾吉泰康(嘉兴)生物科技有限公司
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