Sample preparation and viral detection methods
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsBin Guan, Robert B. Hufnagel
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Abstract
Methods of sample preparation for detection of nucleic acids (such as viral nucleic acids, for example, viral RNA) are provided. In some examples, the methods include contacting a sample with a solution including a buffer containing about 0-0.5 mM EDTA and a chelating resin and incubating the resulting mixture at a temperature of at least about 55° C.
Claims26
A method of preparing a sample for viral nucleic acid detection, comprising:
- contacting the sample with a solution comprising a buffer at pH 4-10, 0-0.5 mM EDTA, and 1-50% chelating resin selected from the group consisting of styrene divinylbenzene copolymer containing paired iminodiacetate ions, iminodiacetate resin SIR-300, and iminodiacetic acid agarose to form a mixture;
- and
- incubating the mixture at a temperature of at least 55° C. for about 1-60 minutes.
A method of preparing a sample for viral nucleic acid detection, comprising:
- (a) contacting the sample with a solution comprising about 10-100% dimethylsulfoxide (DMSO), about 40% DMSO or about 50% DMSO, and 1-50% chelating resin selected from the group consisting of styrene divinylbenzene copolymer containing paired iminodiacetate ions, iminodiacetate resin SIR-300, and iminodiacetic acid agarose to form a mixture;
- and
- incubating the mixture at a temperature of at least 55° C. for about 1-60 minutes;
- or
- (b) contacting the sample with a solution comprising about 0.1-0.5 M urea and 1-50% chelating resin selected from the group consisting of styrene divinylbenzene copolymer containing paired iminodiacetate ions, iminodiacetate resin SIR-300, and iminodiacetic acid agarose to form a mixture;
- and
- incubating the mixture at a temperature of at least 55° C. for about 1-60 minutes;
- or
- (c) contacting the sample with a solution comprising Hanks Balanced Salt Solution (HBSS), 2% fetal bovine serum, and 1-50% chelating resin selected from the group consisting of styrene divinylbenzene copolymer containing paired iminodiacetate ions, iminodiacetate resin SIR-300, and iminodiacetic acid agarose to form a mixture;
- and
- incubating the mixture at a temperature of at least 55° C. for about 1-60 minutes.
A kit comprising one or more containers comprising a solution comprising 10 mM Tris-HCl, 0.1 mM EDTA, and 5% styrene divinylbenzene copolymer containing paired iminodiacetate ions.
A kit comprising one or more containers comprising a solution comprising:
- 10-50% dimethylsulfoxide (DMSO) and 5% styrene divinylbenzene copolymer containing paired iminodiacetate ions;
- or
- 0.1-0.5 M urea and 5% styrene divinylbenzene copolymer containing paired iminodiacetate ions;
- or
- Hanks Balanced Salt Solution (HBSS), 2% fetal bovine serum, and 5% styrene divinylbenzene copolymer containing paired iminodiacetate ions.
Dependencies are inferred from the claim text.