Sequences diagnostic for shrimp pathogens
AssigneeE.I. du Pont de Nemours and Company
InventorsRichard C. Ebersole, Jianzhong Zhang, Mario W. Chen, Christian Peter Lenges
Life of this patent
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Abstract
Primers have been isolated that are diagnostic for the detection of Vibrio harveyi. The primers are based on a portion of the Vibrio harveyi LuxR gene and may be used in primer directed amplification or nucleic acid hybridization assay methods.
Claims19
A pair of Vibrio harveyi diagnostic primer sequences as set forth in SEQ ID NO:1 and SEQ ID NO:2.
A method for detecting the presence of Vibrio harveyi in a sample comprising:
- (i) providing DNA from a sample suspected of containing Vibrio harveyi;
- and
- (ii) probing the DNA with a probe derived from the isolated Vibrio harveyi diagnostic primer sequence as set forth in SEQ ID NO:1 or SEQ ID NO:2 under suitable hybridization conditions;
- wherein the identification of a hybridizable nucleic acid fragment confirms the presence of Vibrio harveyi.
A method for detecting the presence of Vibrio harveyi in a sample comprising:
- (i) providing DNA from a sample suspected of containing Vibrio harveyi;
- and
- (ii) amplifying the DNA with a pair of Vibrio harveyi diagnostic primer sequences as set forth in SEQ ID NO:1 and SEQ ID NO:2 such that amplification products are generated;
- wherein the presence of amplification products confirms the presence of Vibrio harveyi.
A method for quantifying the amount of Vibrio harveyi in a sample comprising:
- (i) providing DNA from a sample suspected of containing Vibrio harveyi;
- (ii) amplifying the DNA with a pair of Vibrio harveyi diagnostic primer sequences as set forth in SEQ ID NO:1 and SEQ ID NO:2 by thermal cycling between at least a denaturing temperature and an extension temperature in the presence of a nucleic acid-binding fluorescent agent or a fluorescently labeled probe;
- (iii) measuring the amount of fluorescence generated by the nucleic acid-binding fluorescent agent or the fluorescently labeled probe during the thermal cycling;
- (iv) determining a cycle threshold number at which the amount of fluorescence generated by the nucleic acid-binding fluorescent agent or the fluorescently labeled probe reaches a fixed threshold value above a baseline value;
- and
- (v) calculating the amount of Vibrio harveyi in the sample by comparing the cycle threshold number determined for the Vibrio harveyi in the sample with a standard curve of the cycle threshold number versus the logarithm of template concentration determined using standard solutions of known concentration.
Dependencies are inferred from the claim text.