Elisa for diagnosis of
An ELISA for the diagnosis of Haemonchus longistipes infection in camels is provided. The ELISA for the diagnosis of Haemonchus longistipes infection in camels includes antibodies raised against a 76 kDa protein isolated from Haemonchus longistipes . The resulting antibodies may be used in assays to detect Haemonchus longistipes infected camels. The assays may be any kind of enzyme-linked immunosorbent assay, or “ELISA”, known to those of skill in the art. The assay using the antibodies raised against a 76 kDa protein isolated from Haemonchus longistipes infected camels may be capable of detecting pre-patent Haemonchus longistipes infection.
1. A method of producing a Haemonchus longistipes ( H. longistipes ) specific antibody, comprising the steps of:
(1) preparing H. longistipes crude extract;
(2) isolating an approximately 76 kDa protein from the H. longistipes crude extract;
(3) purifying the approximately 76 kDa protein by Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-page) and electro-elution;
(4) raising antibodies against the approximately 76 kDa protein in a non-camelid mammal; and
(5) purifying antibodies from the non-camelid mammal, wherein
the step of raising antibodies against the approximately 76 kDa protein further comprises immunization and boosting steps, and
the immunization step comprises injecting the non-camelid mammal with 100 μg of the 76 kDa protein on day 0 and day 14; and wherein the boosting step comprises injecting the non-camelid mammal with 50 μg of the 76 kDa protein on day 49 and 77, and a final injection of 25 μg of the 76 kDa protein on day 196.
2. The method of claim 1 , wherein purifying the approximately 76 kDa protein comprises:
separation of the approximately 76 kDa protein on a SDS-page gel;
excision of the approximately 76 kDa protein;
electrophoretic elution of the approximately 76 kDa protein; and
concentration of the approximately 76 kDa protein.
3. The method of claim 1 wherein the non-camelid mammal is a rabbit.
4. The method claim 1 , further comprising collecting serum from the non-camelid mammal on day 24, day 56, day 84, day 203, or day 210 after the immunization step.
5. The method of claim 1 , wherein the antibodies are purified using affinity chromatography with protein A sepharose.
6. The method of claim 1 , further comprising lyophilizing the H. longistipes specific antibody.
7. A Haemonchus longistipes ( H. longistipes ) specific antibody produced by the method of claim 1 .