Sequences diagnostic for shrimp pathogens
View Patent ↗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.
1. 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 identification of a hybridizable nucleic acid fragment confirms the presence of Vibrio harveyi.
2. The method for detecting the presence of Vibrio harveyi in a sample according to claim 1 wherein the probe derived from the isolated Vibrio harveyi diagnostic primer sequence is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, and the complete complementary sequences thereof.
3. The method according to claim 1 wherein the probe contains a replication inhibiting moiety at the 3′ end.
4. The method according to claim 3 wherein the replication inhibiting moiety is selected from the group consisting of dideoxynuleotides, 3′ deoxynucleotides, a sequence of mismatched nucleosides or nucleotides, 3′ phosphate groups and chemical agents.
5. The method according to claim 4 where-in the 3′ deoxynucleotide is cordycepin.
6. The method according to claim 1 wherein the method further comprises the steps of:
a) treating an environment from which the sample was obtained with a chemical treatment to kill or control the Vibrio harveyi ; and
b) repeating the steps of the method of claim 1 on another sample taken from the environment in order to assess Vibrio harveyi inactivation by the chemical treatment.
7. The method according to claim 1 wherein the method further comprises the step of: treating an environment from which the sample was obtained with a chemical treatment to kill or control the Vibrio harveyi in order to improve health and grow-out of shrimp.