Methods for rapid detection and identification of bioagents in forensic studies
First Claim
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1. A method of identifying a geographic origin of an unknown bioagent in a sample comprising:
- a) contacting nucleic acid from said bioagent with at least one pair of oligonucleotide primers which hybridize to sequences of said nucleic acid, wherein said sequences flank a variable nucleic acid sequence of the bioagent;
b) amplifying said variable nucleic acid sequence to produce an amplification product;
c) determining the molecular mass of said amplification product; and
d) comparing said molecular mass to one or more molecular masses of amplification products obtained by performing steps a)-c) on a plurality of known organisms with known geographic origins, wherein a match identifies said geographic origin of said unknown bioagent.
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Abstract
Method for detecting and identifying unknown bioagents, including bacteria, viruses and the like, by a combination of nucleic acid amplification and molecular weight determination using primers which hybridize to conserved sequence regions of nucleic acids derived from a bioagent and which bracket variable sequence regions that uniquely identify the bioagent. The result is a “base composition signature” (BCS) which is then matched against a database of base composition signatures, by which the bioagent is identified.
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39 Claims
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1. A method of identifying a geographic origin of an unknown bioagent in a sample comprising:
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a) contacting nucleic acid from said bioagent with at least one pair of oligonucleotide primers which hybridize to sequences of said nucleic acid, wherein said sequences flank a variable nucleic acid sequence of the bioagent;
b) amplifying said variable nucleic acid sequence to produce an amplification product;
c) determining the molecular mass of said amplification product; and
d) comparing said molecular mass to one or more molecular masses of amplification products obtained by performing steps a)-c) on a plurality of known organisms with known geographic origins, wherein a match identifies said geographic origin of said unknown bioagent. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A method of identifying a geographic origin of an unknown bioagent in a sample comprising:
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a) contacting nucleic acid from said bioagent with at least one pair of oligonucleotide primers which hybridize to sequences of said nucleic acid, wherein said sequences flank a variable nucleic acid sequence;
b) amplifying said variable nucleic acid sequence to produce an amplification product;
c) determining the base composition of said amplification product; and
d) comparing said base composition to one or more base compositions of amplification products obtained by performing steps a)-c) on a plurality of known organisms with known geographic origins, wherein a match identifies said geographic origin of said unknown bioagent. - View Dependent Claims (21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
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Specification