COMPOSITIONS FOR USE IN IDENTIFICATION OF ADVENTITIOUS VIRUSES
First Claim
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1. A composition, comprising at least one purified oligonucleotide primer pair that comprises forward and reverse primers, wherein said primer pair comprises nucleic acid sequences that are substantially complementary to nucleic acid sequences of two or more different bioagents belonging to the Parvoviridae family, wherein said primer pair is configured to produce amplicons comprising different base compositions that correspond to said two or more different bioagents.
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Abstract
The present invention provides compositions, kits and methods for rapid identification and quantification of adventitious contaminant viruses by molecular mass and base composition analysis.
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Citations
50 Claims
- 1. A composition, comprising at least one purified oligonucleotide primer pair that comprises forward and reverse primers, wherein said primer pair comprises nucleic acid sequences that are substantially complementary to nucleic acid sequences of two or more different bioagents belonging to the Parvoviridae family, wherein said primer pair is configured to produce amplicons comprising different base compositions that correspond to said two or more different bioagents.
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15. The composition of claim 15, wherein said mass modified nucleobase is 5-Iodo-C.
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18. A kit, comprising at least one purified oligonucleotide primer pair that comprises forward and reverse primers that are about 20 to 35 nucleobases in length, and wherein said forward primer comprises at least 70%, at least 80%, at least 90%, at least 95%, or at least 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOS:
- 373-379, and said reverse primer comprises at least 70% sequence identity with a sequence selected from the group consisting of SEQ ID NOS;
380-386.
- 373-379, and said reverse primer comprises at least 70% sequence identity with a sequence selected from the group consisting of SEQ ID NOS;
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19. A method of determining a presence of a parvovirus in at least one sample, the method comprising:
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(a) amplifying one or more segments of at least one nucleic acid from said sample using at least one purified oligonucleotide primer pair that comprises forward and reverse primers that are about 20 to 35 nucleobases in length, and wherein said forward primer comprises at least 70%, at least 80%, at least 90%, at least 95%, or at least 100% sequence identity with a sequence selected from the group consisting of SEQ ID NOs;
373-379, and said reverse primer comprises at least 70% sequence identity with a sequence selected from the group consisting of SEQ ID NOs;
380-386 to produce at least one amplification product; and(b) detecting said amplification product, thereby determining said presence of said parvovirus in said sample. - View Dependent Claims (20, 21, 22, 23)
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24. A method of identifying one or more parvovirus bioagents in a sample, the method comprising:
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(a) amplifying two or more segments of a nucleic acid from said one or more parvovirus bioagents in said sample with two or more oligonucleotide primer pairs to obtain two or more amplification products; (b) determining two or more molecular masses and/or base compositions of said two or more amplification products; and (c) comparing said two or more molecular masses and/or said base compositions of said two or more amplification products with known molecular masses and/or known base compositions of amplification products of known parvovirus bioagents produced with said two or more primer pairs to identify said one or more parvovirus bioagents in said sample. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43)
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44. A system, comprising:
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(a) a mass spectrometer configured to detect one or more molecular masses of amplicons produced using at least one purified oligonucleotide primer pair that comprises forward and reverse primers, wherein said primer pair comprises nucleic acid sequences that are substantially complementary to nucleic acid sequences of two or more different parvovirus bioagents; and (b) a controller operably connected to said mass spectrometer, said controller configured to correlate said molecular masses of said amplicons with one or more parvovirus bioagent identities. - View Dependent Claims (45, 46, 47, 48, 49)
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50. A composition comprising an isolated primer 15-35 bases in length selected from the group consisting of SEQ ID NOs 373-386.
Specification