Nucleic acid decontamination methods
Inventors
KUTYAVIN, Alex I. • Lund, Kevin P. • Nanassy, Oliver Z. • Gall, Alexander A. • BRABANT, William
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Assignees
Member
CepheidCepheid is a global leader in molecular diagnostics, dedicated to improving healthcare by developing, manufacturing, and marketing automated, easy-to-use molecular systems and tests. Their mission is to provide rapid, accurate, and actionable genetic testing for a wide range of infectious diseases, oncology, and human genetics. Cepheid's flagship GeneXpert System delivers scalable, sample-to-answer PCR testing for institutions of any size, supporting both centralized and decentralized care. The company is committed to expanding access to high-quality diagnostics worldwide, supporting public health initiatives, driving innovation in molecular testing, and advancing sustainability and responsible business practices.
Cepheid is a global leader in molecular diagnostics, dedicated to improving healthcare by developing, manufacturing, and marketing automated, easy-to-use molecular systems and tests. Their mission is to provide rapid, accurate, and actionable genetic testing for a wide range of infectious diseases, oncology, and human genetics. Cepheid's flagship GeneXpert System delivers scalable, sample-to-answer PCR testing for institutions of any size, supporting both centralized and decentralized care. The company is committed to expanding access to high-quality diagnostics worldwide, supporting public health initiatives, driving innovation in molecular testing, and advancing sustainability and responsible business practices.
Publication Number
US-11819584-B2
Publication Date
2023-11-21
Expiration Date
Abstract
Methods and cleaning compositions for reduction of nucleic acid contamination on surfaces, in air, and in solutions using modified pectin are provided.
Core Innovation
Methods and cleaning compositions for reduction of nucleic acid contamination on surfaces, in air, and in solutions using modified pectin are provided. The invention provides methods of reducing nucleic acid contamination by contacting a contaminated surface with a composition comprising a modified pectin, wherein the modified pectin comprises a plurality of amino groups, for example an amidated pectin. The decontaminating agents can be used in the form of a solution, a suspension, or can be bound to a solid support.
The background identifies that nucleic acid amplification-based techniques are highly sensitive and vulnerable to contamination, with accumulation of amplicons in the laboratory environment being a major source of cross-contamination. Previously reported decontamination methods typically employ difficult to handle, corrosive reagents and do not consistently and effectively degrade amplifiable nucleic acids or selectively remove nucleic acids from solutions while keeping other biomolecules intact, creating a need for improved, inexpensive nucleic acid decontamination methods that employ easy to use, stable reagents and that are compatible with a wide variety of substrates and surfaces.
The amidated pectin compounds disclosed herein are believed to facilitate flocculation of nucleic acids, the process by which individual molecules of nucleic acids aggregate or precipitate into small particles when bound to the modified pectins, thus rendering the nucleic acids unsuitable for amplification. The invention provides use of amidated pectins in solutions, on solid supports, and in air-contacting applications to render contaminating nucleic acids unamplifiable and to reduce nucleic acid contamination.
Claims Coverage
The independent claims define three inventive features directed to methods for reducing nucleic acid contamination on surfaces, in solutions, and in air using amidated pectins.
Surface decontamination using an amidated pectin
Contacting a surface contaminated with a nucleic acid with a composition comprising an amidated pectin comprising one or more monomeric units represented by the formulas disclosed in the specification, wherein the amidated pectin comprises a plurality of amino groups.
Solution decontamination using an amidated pectin covalently bound to a solid support
Contacting a solution contaminated with a nucleic acid with a solid support comprising an amidated pectin covalently bound thereto, wherein the amidated pectin comprises one or more monomeric units represented by the formulas disclosed in the specification.
Air decontamination of aerosolized nucleic acid using an amidated pectin
Contacting air contaminated with aerosolized nucleic acid with a composition comprising an amidated pectin comprising one or more monomeric units represented by the formulas disclosed in the specification, including passing air through a solution, suspension, or a filter comprising the amidated pectin.
In summary, the independent claims cover use of amidated pectins defined by one or more specified monomeric units and comprising multiple amino groups for (1) contacting contaminated surfaces, (2) use on solid supports to decontaminate solutions, and (3) contacting aerosolized nucleic acids in air; the amidated pectins can be applied as solutions, suspensions, or covalently bound to supports.
Stated Advantages
Render nucleic acids unsuitable for amplification by facilitating flocculation of nucleic acids, thereby aggregating or precipitating nucleic acids and making them unamplifiable.
Do not react with components of molecular diagnostics tests with the release of toxic byproducts, unlike bleach or other oxidants.
Decontaminating compositions can be stable upon storage at room temperature.
Can bind minor DNA contaminants while allowing amplification of desired target DNA to take place, as demonstrated by examples showing contaminant binding without preventing target amplification.
Documented Applications
Reduction of nucleic acid contamination on surfaces, including surfaces of instruments, laboratory bench surfaces, and laboratory pipettes, by contacting the surface with compositions comprising amidated pectins.
Reduction of nucleic acid contamination in solutions by contacting the solution with solid supports comprising amidated pectins covalently bound thereto, including use of magnetic beads, glass beads, polystyrene beads, filters, and similar supports.
Reduction of aerosolized nucleic acid contamination in air by contacting contaminated air with compositions comprising amidated pectins, including passing air through aqueous solutions or filters comprising amidated pectins and use in scrubbers or air filters.
Use to remove amplicon contamination and to prevent contamination of molecular diagnostics laboratory surfaces, instrumentation, and equipment, including adding decontaminating solution post-amplification to render amplification products substantially unamplifiable and use in automated cartridges.
Incorporation of amidated pectins into wipes, cloths, sponges, pads, filters, or impregnated materials for decontamination purposes.
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