BIOAGENT DETECTION METHODS

• US 20110318821A1
• Filed: 03/08/2010
• Published: 12/29/2011
• Est. Priority Date: 03/08/2009
• Status: Active Grant

• Current Assignee: Ibis Biosciences Incorporated
• Original Assignee: Ibis Biosciences Incorporated
• Inventors: Hofstadler, Steven A.
• Granted Patent: US 9,719,083 B2
• Time in Patent Office: Days
• US Class Current: 435/287.2
• CPC Class Codes: C12N 15/101 by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

The present invention relates the use of broad range primer (e.g., as broad range capture olignucleotides) immobilized in a SCODA method gel to allow, for example, selective concentration of target nucleic acids. Such concentrated target nucleic acids may, for example, be: i) eluted from the gel and analyzed (e.g., by broad range primer methods); ii) subject to in situ (e.g., in gel) PCR methods; and/or iii) analyzed in the gel (e.g., by fluorescent detection methods).

Claims

1. A method of selectively concentrating target nucleic acids from a sample comprising;

a) providing;

i) a device for performing SCODA purification, wherein said device comprises a gel,

ii) a plurality of broad range survey primers, wherein said broad range survey primers are immobilized in said gel,

iii) a sample comprising non-target nucleic acid, contaminants, and possibly target nucleic acid; and

b) introducing said sample into said device under conditions such that any target nucleic acid in said sample is separated from said contaminants and is captured by said at least some of said broad range survey primers.

2. The method of claim 1, wherein said target nucleic acid is separated from said non-nucleic acid in said sample.

3. The method of claim 1, wherein said target nucleic acid is eluted from said gel to generate a preparation substantially free of non-target nucleic acid and contaminants.

4. The method of claim 3, wherein said target nucleic acid in said preparation is subjected to amplification using primers.

5. The method of claim 4, wherein said primers are the same as at least some of said plurality of primers.

6. The method of claim 1, wherein said target nucleic acid is amplified in said gel employing said plurality of broad range survey primers.

7. The method of claim 1, wherein said capture of said target nucleic acid is detected in said gel.

8. An instrument for selectively concentrating target nucleic acids from a sample comprising;

a) a device for performing SCODA purification, wherein said device comprises a gel, and

b) a plurality of broad range survey primers, wherein said broad range survey primers are immobilized in said gel.

9. A system or kit comprising:

a) a gel suitable for use in SCODA purification; and

b) a plurality of broad range survey primers.

10. The system or kit of claim 9, further comprising a component for generating an electric field.