Sorbents for extraction and stabilization of nucleic acids
Inventors
Lin, Baochuan • White, Brandy J. • Melde, Brian
Assignees
Publication Number
US-10077465-B2
Publication Date
2018-09-18
Expiration Date
2036-09-13
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Abstract
Mesoporous sorbents are effective for storing and transporting nucleic acids. In particular, two ethane-bridged silica sorbents with amine functionalities are particularly effective and capable of binding nucleic acids for storage.
Core Innovation
The invention relates to improved methods and materials applicable to the capture of nucleic acid targets from complex matrices offering subsequent stabilization of the targets in the absence of refrigeration. It focuses on mesoporous ethane-bridged silica sorbents with amine functionalities, particularly two sorbents named N5 and CuEDA, which are highly effective in binding and stabilizing nucleic acids for storage and transport.
The problem being solved is the need for preservation of nucleic acid integrity during sample collection, transportation, processing, and storage without reliance on freezing methods, which are not practical for routine specimen processing or field conditions. RNA is particularly labile and prone to degradation by RNases and environmental conditions, necessitating methods that preserve nucleic acids at room temperature in a cost-effective and reliable manner.
The disclosed solution leverages mesoporous silica sorbents bearing chemical functionalities that allow adsorption and stabilization of nucleic acids, reducing their degradation and enabling storage and transport under ambient or elevated temperatures. The sorbents may be modified or combined with stabilizing groups such as trehalose or bovine serum albumin to enhance protection. The sorbents also allow elution of nucleic acids using mild buffers after an initial detergent wash, facilitating recovery for downstream molecular diagnostics.
Claims Coverage
The patent includes one independent claim detailing a method of storing nucleic acids using specific mesoporous silica sorbents and related procedural features.
Method of storing nucleic acid with mesoporous ethane-bridged silica sorbent
A method comprising contacting nucleic acid with a mesoporous ethane-bridged silica sorbent comprising both N5 and CuEDA sorbents, causing the nucleic acid to releasably bind to the sorbent.
Pre-washing sorbents with detergent to improve binding and elution
A preparatory step of washing the sorbent with a detergent, such as 1% Triton X-100, prior to contacting nucleic acid to enhance binding and facilitate nucleic acid recovery.
Elution step for nucleic acid recovery
A step of eluting the nucleic acid from the sorbent after binding using mild buffer conditions, enabling recovery of the nucleic acid for further use.
Contacting nucleic acid in complex matrices
The method applies to contacting nucleic acid within complex matrices, including lysed cells, allowing direct binding without purification.
Nucleic acid comprising RNA
The method is applicable specifically to nucleic acids including RNA targets.
Association of sorbent with stabilizers
The sorbent can be associated with stabilizers, such as bovine serum albumin, trehalose, or glucosamine, to enhance nucleic acid stabilization during storage.
The claims cover a method of nucleic acid storage involving contacting nucleic acid with mesoporous silica sorbents N5 and CuEDA, including preparatory detergent washing, elution recovery steps, application to complex matrices, RNA targets, and association with stabilizers for improved preservation.
Stated Advantages
Mesoporous silica sorbents provide effective capture and stabilization of nucleic acids at room temperature and elevated temperatures (up to 37°C).
The sorbents allow binding and stabilization of nucleic acids directly from complex solutions such as lysed cells, reducing the need for purification steps.
Pre-conditioning the sorbents with detergent improves target binding, stability, and recovery yield during elution.
Stabilizers covalently linked or mixed with the sorbents (e.g., trehalose, bovine serum albumin) enable long-term preservation without downstream contamination concerns.
The materials are reusable and environmentally friendly, enabling cost-effective sample handling under field conditions without refrigeration.
Documented Applications
Storage and transport of nucleic acids from complex biological samples, including lysed cells, for subsequent molecular diagnostic assays.
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