Aptamer Inactivation of Nucleases in Biological Solutions, Reagents, and Kits Used for Sample Collection, Nucleic Acid Testing, Isolation, and Genomic Characterization

A method for utilization of anti-nuclease aptamers in sample/specimen collection medium for inactivation of nucleases (RNase and DNase) from collected biological matrices where preservation and stabilization of RNA and DNA are critical to downstream nucleic acids detection and characterization processes, the method comprising the steps of:

• preparing a collection medium comprised of reagents that include at least an aptamer and a membrane lysis detergent;
• collecting a biological sample fluid or swab comprising biological matrices;
• adding, combining and mixing the biological sample and the collection medium into a closed collection container to form a collection mixture;
• performing at least partial lysis of the collected biological sample to release nucleic acids and proteins from cells in the biological matrices;
• allowing the nucleases (RNase and DNase) to bind to the aptamer in the collection mixture;
• and
• inactivating the nucleases to enhance stability and preservation of RNA and DNA from the biological sample;
• whereby the method provides a multi-use collection system suitable for testing and characterizing nucleic acids, proteins and antigens with or without the use of prefatory nucleic acid extraction or other purification steps.

A formulation for a sample collection medium containing an aptamer or collection of aptamers that bind with high affinity and subsequently inactivate nucleases in a collected sample:

• the collection medium contains at least one buffer, at least one sugar, at least one salt, at least one detergent, at least one chelating agent, and at least one PCR enhancer, the combined collection medium buffered between 7 and 7.4 with HCl;
• the medium contains TRIS, sucrose, sodium acetate, Tween-20, EDTA, betaine, and glycerol, it includes, but is not limited to, the aptamers defined by SEQ ID NO:
• 1-SEQ ID NO:
• 5 described herein for RNase inactivation;
• the aptamers inactivate RNase at 8,800-fold to 10,200-fold and preserve collected samples at ambient and elevated ambient temperatures up to 30° C.;
• the collection medium containing a collected sample can be used with nucleic acid extraction or used directly without extraction in molecular tests, including PCR;
• the collection medium containing collected samples can be utilized for multiple diagnostic, detection, and characterization methods for molecular nucleic acid tests and genomic sequencing applications, ELISA, protein, lateral flow, and rapid antigen tests;
• wherein the collection medium containing collected sample can be utilized for multiple diagnostic, detection, and characterization methods molecular nucleic acids tests and genomic sequencing applications, ELISA, protein, lateral flow, and rapid antigen tests.