Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Detection module and method

A detection module, nucleic acid sequence technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as inability to perform on-site detection, time-consuming, and little universal detection.

Inactive Publication Date: 2018-11-16
CHINA AGRI UNIV
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are many types of food safety risk factors, and the connection between different types of risk factors is getting closer. However, it is still a technical problem to realize the simultaneous detection of different types of risk factors.
Traditional detection methods rely on large-scale equipment or immunoassays, etc., which are usually time-consuming and cannot be detected on-site
However, some biological assays are often highly selective for the target substances to be detected, and they are also rarely available for universal detection.

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
  • Detection module and method
  • Detection module and method
  • Detection module and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Example 1, Nucleotide sequences used in EXPAR tandem digestion HCR detection module and design method thereof

[0094] (1) Nucleotide sequence used by EXPAR

[0095] The nucleotide sequences used in the EXPAR designed in this embodiment are shown in Table 1.

[0096] Table 1

[0097]

[0098] (2) Nucleotide sequences of traditional HCR and digested HCR

[0099] The nucleotide sequences used in traditional HCR and digested HCR designed in this example are shown in Table 2.

[0100] Table 2

[0101]

[0102] In Table 2, hairpin H1 is hairpin 1, hairpin H2 is hairpin 2, which is used for traditional HCR; D-H1 is digestion hairpin 1, and D-H2 is digestion hairpin 2, which is used for digestion type HCR.

[0103] (3) Design method of EXPAR serial digestion type HCR general module

[0104] In this embodiment, sequence X is used to represent the target sequence to be detected. In the actual detection process, the primer sequence can be designed according to the tar...

Embodiment 2

[0127] Embodiment 2, establishment and optimization of EXPAR reaction system, reaction conditions

[0128] (1) Establishment of EXPAR reaction system and reaction conditions

[0129] The EXPAR reaction system established in this example is divided into two systems, A and B, as shown in Table 4 and Table 5 respectively. The established EXPAR reaction system was verified by real-time PCR method.

[0130] System A is shown in Table 4.

[0131] Table 4

[0132]

[0133]

[0134] System B is shown in Table 5.

[0135] table 5

[0136]

[0137] EXPAR reaction conditions:

[0138] Add the samples except SYBR Green I in system A, mix well, put in a reaction tube, and heat at 55°C for 5min. After taking it out, add SYBR Green I (avoid light as much as possible). Add system B to system A, centrifuge to mix the reactants, and place them in a real-time quantitative PCR instrument for reaction. The design program is 55°C, 250 cycles.

[0139] Experimental results such as f...

Embodiment 3

[0148] Embodiment 3, establishment and optimization of digested HCR

[0149] (1) Self-assembly effect of digested HCR, verification of priming efficiency and establishment of self-assembly reaction conditions

[0150] The hairpin H1 and hairpin H2 used for traditional HCR in Table 2 of Example 1 were used as the control group (hairpin H1 is abbreviated as H1 in the following description, and hairpin H2 is abbreviated as H2), and D-H1 and D- H2 is used as the experimental group, when the concentration of each hairpin structure is 100nM, the promoter concentration in Table 2 of Example 1 is 10nM, and the self-assembly reaction time is 30min, 1h, 90min, 2h, 150min, 3h, 4h respectively , the self-assembly effect and priming efficiency of the experimental group and the control group.

[0151] 1) Self-assembly reaction system: each of the two hairpin structures is 100nM, the promoter is 10nM, and the reaction buffer (8.0mMNa 2 HPO 4 ,2.5mM NaH 2 PO 4 ,2.0mM MgCl 2 , and 0.15mM...

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

PropertyMeasurementUnit
Sensitivityaaaaaaaaaa
Login to View More

Abstract

The invention provides a detection module and method. The detection module and method are established on the basis of an EXPAR (exponential amplification reaction) tandem digestion type HCR (hybridization chain reaction). When a to-be-detected object contains a target substance, a large quantity of digestion single strands are formed by amplification with the established module and method for an EXPAR, detection signals of the target substance are amplified, then the formed digestion single strands are used for digesting a modified digestion type HCR product, and the detection signals of the target substance are converted. In one specific embodiment, supermolecular double-strand concatemers are modified and upgraded by modifying hairpin structures used in the HCR, so that not only can DNAdouble strands be self-assembled by nucleic acid hairpins, but also the nucleic acid hairpins can be digested efficiently by nucleic acid single strands. Dual induction of signals is realized. In another embodiment, when the designed and modified digestion type hairpin sequences and digestion sequences are applied to practical detection, detection sensitivity is high, and 1nM X substrate can be detected at lowest.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to a detection module and a method. Background technique [0002] The source of food safety risk factors can be roughly divided into biological source safety risk factors and non-biological source safety risk factors. Biological source safety risk factors are more typical, such as food-borne pathogenic bacteria and microRNA with important biological significance in the study of toxicity mechanism; non-biological source food safety risk factors such as heavy metals in the environment. In addition, there are many types of food safety risk factors, and the links between different types of risk factors are getting closer. However, it is still a technical problem to realize the simultaneous detection of different types of risk factors. Traditional detection methods rely on large-scale equipment or immunoassays, etc., which are usually time-consuming and c...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/682
CPCC12Q1/682C12Q2525/301
Inventor 许文涛罗云波苗苗杜再慧田晶晶梁志宏黄昆仑
Owner CHINA AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com