Unlock instant, AI-driven research and patent intelligence for your innovation.

Cracking binding solution based on paramagnetic particle method pathogen nucleic acid extraction and product and application thereof

A technology for lysing binding solution and pathogens, applied in the biological field, can solve the problems of strong corrosion of the sealing membrane of deep well plates, product leakage, and complicated operation, so as to shorten the nucleic acid extraction time, reduce the possibility of leakage, and simplify the operation. Effects of nucleic acid extraction steps

Active Publication Date: 2021-11-26
SHANGHAI BIOGERM MEDICAL TECH CO LTD
View PDF7 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, conventional magnetic bead nucleic acid extraction reagents have been able to cooperate with automated instruments. Among them, pre-loaded nucleic acid extraction reagents in deep-well plates are a relatively common form. However, since many reagents contain ethanol or isopropanol, etc. Organic solvents have a strong corrosive effect on the sealing film of deep-well plates, which may easily cause product leakage, and too much organic solution will also have some adverse effects on samples with high protein content
In addition, the operation of most nucleic acid extraction reagents is cumbersome at present, and usually includes components such as lysis solution, binding solution, deproteinization solution, washing solution, and eluent. Some nucleic acid extraction reagents also need to add proteinase K and RNA precipitation aids. and other additional components, even with automated instruments, there are still many manual operations, and the multi-step washing makes the entire operation time about 20min

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
  • Cracking binding solution based on paramagnetic particle method pathogen nucleic acid extraction and product and application thereof
  • Cracking binding solution based on paramagnetic particle method pathogen nucleic acid extraction and product and application thereof
  • Cracking binding solution based on paramagnetic particle method pathogen nucleic acid extraction and product and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] A kind of lysing binding solution based on magnetic bead method pathogenic nucleic acid extraction, is made of following components: 100-140mM Tris-HClpH8.5, 3.0M guanidine isothiocyanate, 3 (w / v)%NP-40, 0% ( w / v) SDS, 5% (w / v) PEG6000, 3% (w / v) Triton X-100 and 10-30 mM disodium edetate, water as solvent. Optimize the concentration ratio of Tris-HCl and disodium edetate, the specific collocation is as follows:

[0055] Table 1 Buffer system optimization scheme

[0056]

[0057] In this example, the throat swab sample simulated by the new crown pseudovirus is used as the nucleic acid extraction sample, and the sample matrix is ​​the sampling kit of different manufacturers (manufacturer 1, manufacturer 2, and manufacturer 3), and the final extracted Ct value is used as the judgment of the nucleic acid extraction effect standard.

[0058] The nucleic acid extraction step employs the above-mentioned steps. Wherein, the washing liquid adopts: 80% ethanol.

[0059] A ...

Embodiment 2

[0065] A lysing binding solution based on magnetic bead method pathogenic nucleic acid extraction, consisting of the following components: 120mM Tris-HClpH8.5, 3.0-3.8M guanidine isothiocyanate, 3 (w / v)% NP-40, 0% ( w / v) SDS, 5-15% (w / v) PEG6000, 3% (w / v) Triton X-100 and 20 mM disodium edetate, water as solvent. Wherein the concentration of guanidine isothiocyanate and PEG is optimized, the specific preparation method is as follows:

[0066] Table 3 combined system optimization scheme

[0067] serial number Guanidine isothiocyanate (M) PEG (w / v%) recipe one 3 5 recipe two 3 10 Recipe three 3 15 Recipe four 3.4 5 Recipe five 3.4 10 Recipe six 3.4 15 Recipe seven 3.8 5 Recipe Eight 3.8 10 Recipe Nine 3.8 15

[0068] In this example, the throat swab sample simulated by the new coronavirus is used as the nucleic acid extraction sample, and the sample matrix is ​​the pathogen preservation solution, a...

Embodiment 3

[0075] A lysing binding solution based on magnetic bead method for pathogenic nucleic acid extraction, consisting of the following components: 120mM Tris-HCl pH8.5, 3.4M guanidine isothiocyanate, 3-9 (w / v)% NP-40, 0 - 0.5% (w / v) SDS, 10% (w / v) PEG6000, 3-9% (w / v) Triton X-100 and 20 mM disodium edetate, water as solvent. The concentration of NP-40, Triton-X-100 and SDS is optimized, and the specific preparation method is as follows:

[0076] Table 5 cracking system optimization

[0077] serial number NP-40(w / v%) Triton-X-100(w / v%) SDS(w / v%) recipe one 3 3 0 recipe two 3 6 0.25 Recipe three 3 9 0.5 Recipe four 6 3 0 Recipe five 6 6 0.25 Recipe six 6 9 0.5 Recipe seven 9 3 0 Recipe Eight 9 6 0.25 Recipe Nine 9 9 0.5

[0078] In this example, throat swab samples simulated by the new coronavirus and Staphylococcus aureus cultures were used as nucleic acid extraction samples, and the sample...

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 relates to the technical field of biology, and particularly provides a cracking binding solution based on paramagnetic particle method pathogen nucleic acid extraction and a product and application thereof. The cracking binding solution comprises the following components: 100 to 140 mM of Tris-HCl with the pH value of 8.0-9.0, 3.0 to 3.8 M of guanidine isothiocyanate, 3 to 9 w / v percent of NP-40, 0 to 0.5 w / v percent of SDS, 5 to 15 w / v percent of PEG6000, 3 to 9 w / v percent of Triton X-100 and 10 to 30 mM of disodium ethylene diamine tetraacetate. As a whole, the cracking binding liquid can realize two processes of to-be-detected sample cracking and nucleic acid and magnetic bead binding, and a high-quality nucleic acid sample which does not inhibit subsequent PCR detection can be extracted without using protease K, an RNA settling aid and the like. Corrosivity is avoided, meanwhile, the nucleic acid extraction step is simplified, and the nucleic acid extraction time is shortened.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a lysing binding solution based on magnetic bead method for nucleic acid extraction of pathogens, as well as its products and applications. Background technique [0002] With the development of diagnostic technology, molecular diagnostic technology has gradually become the main means of diagnosing diseases in human daily life, and molecular diagnostic technology based on nucleic acid information is an important means among them. Therefore, nucleic acid extraction before molecular diagnosis is not only a necessary step, but also Its requirements are also constantly increasing. [0003] A common method for nucleic acid extraction is the magnetic bead method. At present, conventional magnetic bead nucleic acid extraction reagents have been able to cooperate with automated instruments. Among them, pre-loaded nucleic acid extraction reagents in deep-well plates are a relatively common fo...

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): C12N15/10
CPCC12N15/1013
Inventor 沈子明赵百慧
Owner SHANGHAI BIOGERM MEDICAL TECH CO LTD