Methods for rapid identification of pathogens in humans and animals
First Claim
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1. A method of identifying one or more etiologic agents of disease in a sample comprising the steps of:
- amplifying two or more segments of a nucleic acid from said one or more etiologic agents in said sample with two or more primer pairs to obtain two or more amplification products;
determining base compositions of said two or more amplification products, wherein said base compositions identifies the number of A residues, C residues, T residues, G residues, U residues, analogues thereof and mass tag residues thereof; and
identifying said one or more etiologic agents in said sample by comparing said base compositions with known base compositions of known etiologic agents produced with said two or more primer pairs.
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Abstract
The present invention provides methods of: identifying pathogens in biological samples from humans and animals, resolving a plurality of etiologic agents present in samples obtained from humans and animals, determining detailed genetic information about such pathogens or etiologic agents, and rapid detection and identification of bioagents from environmental, clinical or other samples.
383 Citations
119 Claims
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1. A method of identifying one or more etiologic agents of disease in a sample comprising the steps of:
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amplifying two or more segments of a nucleic acid from said one or more etiologic agents in said sample with two or more primer pairs to obtain two or more amplification products; determining base compositions of said two or more amplification products, wherein said base compositions identifies the number of A residues, C residues, T residues, G residues, U residues, analogues thereof and mass tag residues thereof; and identifying said one or more etiologic agents in said sample by comparing said base compositions with known base compositions of known etiologic agents produced with said two or more primer pairs. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31)
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32. A method of identifying one or more bioagents in a sample comprising the steps of:
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amplifying two or more segments of a nucleic acid from said one or more of bioagents in said sample with two or more primer pairs to obtain two or more amplification products; determining base compositions of said two or more amplification products, wherein said base compositions identifies the number of A residues, C residues, T residues, G residues, U residues, analogues thereof and mass tag residues thereof; and identifying said one or more bioagents in said sample by comparing said base compositions with known base compositions of known bioagents produced with said two or more primer pairs. - View Dependent Claims (33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59)
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60. A method of identifying one or more bioagents in a sample comprising the steps of:
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amplifying two or more segments of a nucleic acid from said one or more of bioagents in said sample with two or more primer pairs to obtain two or more amplification products, wherein each of the primer pairs hybridizes to conserved regions of the nucleic acid that flank a variable region; determining base compositions of said two or more amplification products, wherein said base compositions identifies the number of A residues, C residues, T residues, G residues, U residues, analogues thereof and mass tag residues thereof; and identifying said one or more bioagents in said sample by comparing said base compositions with a database containing known base compositions of known bioagents produced with said two or more primer pairs. - View Dependent Claims (61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90)
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91. A method of identifying one or more bioagents in a sample comprising the steps of:
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amplifying two or more segments of a nucleic acid from said one or more of bioagents in said sample with two or more primer pairs to obtain two or more amplification products, wherein each of the primer pairs hybridizes to conserved regions of the nucleic acid that flank a variable region; determining the mass of said two or more amplification products via mass spectrometry, without sequencing; and comparing said masses with a database containing known masses of known bioagents produced with said two or more primer pairs to identify said one or more bioagents in said sample. - View Dependent Claims (92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119)
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Specification