Multiplex RT-PCR/PCR for simultaneous detection of bovine coronavirus, bovine rotavirus, Cryptosporidium parvum, and Escherichia coli
AssigneeBoard of Trustees of Michigan State University
InventorsRoger Maes, Annabel Wise
Life of this patent
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Abstract
The present invention provides a multiplex RT-PCR/PCR method, which enables in a single assay the simultaneous detection of any combination of bovine rotavirus, bovine coronavirus, Cryptosporidium parvum, and optionally, Escherichia coli strains producing K99 pili or heat-stable enterotoxin STa.
Claims12
1-11. (canceled)
A kit for simultaneous screening of a fecal sample for at least two pathogens selected from the group consisting of bovine coronavirus, bovine rotavirus, and Cryptosporidium parvum comprising in three or more containers:
- (a) a first oligonucleotide primer pair which hybridizes to opposite strands of a target nucleic acid sequence of Cryptosporidium parvum;
- (b) a second oligonucleotide primer pair which hybridizes to opposite strands of a target nucleic acid sequence of bovine coronavirus;
- and
- (c) a third oligonucleotide primer pair which hybridizes to opposite strands of a target nucleic acid sequence of bovine rotavirus, wherein each primer pair flanks its target nucleic acid sequence for PCR amplification of the target nucleic acid sequence.
A primer pair comprising an upstream primer having the nucleic acid sequence in SEQ ID NO:
- 3 and a downstream primer having the nucleic acid sequence in SEQ ID NO:4.
A primer pair comprising an upstream primer having the nucleic acid sequence in SEQ ID NO:
- 3 and a downstream primer having the nucleic acid sequence in SEQ ID NO:4.
Dependencies are inferred from the claim text.