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Background   Haemonchus contortus  ( H. contortus ) is one of the most important parasites that cause huge economic losses to small ruminant industry worldwide. Effective prognosis and treatment depend upon the early diagnosis of  H. contortus  infection. To date, no widely-approved methods for the identification of prepatent  H. contortus  infection are available to identify prepatent  H. contortus  infection properly. The aim of this study was to evaluate the diagnostic potential of recombinant cold shock  H. contortus  protein (rHc-CS) during early and late infections of  H. contortus  in goat.    Results  Purified rHc-CS exhibited a clear band, with a molecular weight about 38 kDa.  H. contortus  eggs were not detected by fecal egg count technique from feces collected at 0 to 14 days post infection (D.P.I). However, eggs were detected at 21, 28 and 35 D.P.I. Hence, results of immunoblotting assay showed specific anti rHc-CS antibody detection in all goat sera collected at early stage (14 D.P.I) and late stage (21–103 D.P.I) of  H. contortus  infection. Furthermore, no cross reactivity was observed against  Trichinella spiralis ,  Fasciola hepatica  and  Toxoplasma gondii  or uninfected goats. Among several evaluated rHc-CS indirect-ELISA format variables, favorable antigen coating concentration was found 0.28 μg/well at 37 °C 1 h and overnight at 4 °C. Moreover, optimum dilution ratio of serum and rabbit anti-goat IgG was recorded as 1:100 and 1:4000, respectively. The best blocking buffer was 5% Bovine Serum Albumin (BSA) while the best time for blocking, serum incubation and TMB reaction were recorded as 60, 120 and 10 min, respectively. The cut-off value for positive and negative interpretation was determined as 0.352 (OD 450 ). The diagnostic specificity and sensitivity of the rHc-CS, both were recorded as 100%.    Conclusion  These results validated that rHc-CS is a potential immunodiagnostic antigen to detect the specific antibodies during early and late  H. contortus  infections in goat.

bmc veterinary research

Recombinant cold shock domain containing protein is a potential antigen to detect specific antibody during early and late infections of Haemonchus contortus in goat