Comparison of Fecal Egg Counts and ELISA for the Diagnosis of Dicrocoelium Dendriticum Infection

Naeemipour, Mohsen; Hashemitabar, Gh.R.; Dastjerdi, Kazem; Mojaver, M. Jamshidian; Mohammadi, Hamidreza

doi:10.1515/pjvs-2016-0072

Cited by 12 publications

(9 citation statements)

References 20 publications

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“…Fecal egg examination is a routine procedure, which is not able to indicate the precise level of infection in the early stages. Serological methods have become important for the diagnosis of parasites as they are more accurate in comparison with conventional egg examination and can be used to quickly diagnose infection [16]. Among these methods, ELISA has been most extensively studied as it allows the simultaneous and quick examination of a large number of samples.…”

Section: Discussionmentioning

confidence: 99%

Adhesion-Regulating Molecule from Haemonchus contortus: Potential Antigen for Diagnosis of Early Infection in Goats

et al. 2019

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Haemonchus contortus, a blood-sucking nematode of ruminants, causes large economic losses worldwide. Diagnosis of infection mainly depends on the evaluation of clinical signs and fecal examination. However, this has limitations for the diagnosis of early or light infections, where serological diagnosis seems to be more accurate and reliable. In this study, the recombinant H. contortus adhesion-regulating molecule protein (rHCADRM) was expressed and purified, and its diagnostic potential was evaluated. Serum samples from goats experimentally infected with H. contortus (n = 5) were collected at 0 (before infection, negative control), 7, 14, 21, 35, 49, 63, 85, and 103 days post-infection (DPI). The reactions between rHcADRM and goat serum were tested using Western blot (WB) analysis. The results show that rHcADRM can be recognized in the serum as early as 14 DPI, and the antibody against rHcADRM in infected goat could be maintained for over 89 days. No reaction was found between rHcADRM and antibodies against Trichinella spiralis, Fasciola hepatica, or Toxoplasma gondii. An indirect enzyme-linked immune sorbent assay (ELISA) was developed based on rHcADRM. The optimal coating antigen (279 ng of rHcADRM/well) and serum dilutions (1:50) were determined by checkerboard titration. A total of 64 serum samples, including 32 from H. contortus infection goats and 32 from helminth-free goats, were used to determine the positive (0.362) and negative (0.306) cut-off values for the ELISA. The results show this serological diagnosis method is highly sensitive (90.6%) and specific (93.75%). The coefficient of variation within run and between runs was less than 11%. To apply this indirect ELISA during field examination, 51 serum samples were randomly collected from goat farms and tested using this method. The result showed that 19.6% (10/51) of goats were infected with H. contortus, which was 100% consistent with the necropsy result, higher than that of fecal examination (15.7%, 8/51). These results indicate that rHcADRM could be a potential antigen for diagnosis of H. contortus infection in goats.

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Section: Discussionmentioning

confidence: 99%

Adhesion-Regulating Molecule from Haemonchus contortus: Potential Antigen for Diagnosis of Early Infection in Goats

et al. 2019

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“…To control this infection efficiently, early diagnosis of the infection is inevitable as last two larval stages of this parasite feed on blood that loss up to one fifth of the total circulating erythrocyte volume from young animals and may cause fatal anemia [10]. Previous study reported the serological methods for diagnosis of Dicrocoelium dendriticium because these methods are more accurate in comparison to FEC test [25].…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2020

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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.

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“…Early diagnosis of the infection is crucial for effective control of H. contortus infection because last two larval stages of this parasite feed on blood and loss up to one fifth of the total circulating erythrocyte volume from young animals and may cause fatal anemia [10]. The serological methods become important for the diagnosis of parasite as they are more accurate in comparison to EPG test and they are able to diagnose infection in a short time [25].…”

Section: Discussionmentioning

confidence: 99%

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

Aimulajiang

Memon

Hasan

et al. 2019

Preprint

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Background: Haemonchus contortus (H. contortus) is the most abundant nematode causing haemonchosis with major economic losses to the small ruminant industry farming worldwide. Effective prognosis and treatment depend upon the early diagnosis of H. contortus infection. To date, no widely-approved methods are available to identify prepatent H. contortus infection. 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. Fecal egg count technique was unable to detect H. contortus eggs in feces collected between 0 and 14 days post infection. Eggs were detected at 21, 28 and 35 days post infection. However, Specific anti rHc-CS antibodies were detectable in sera of all infected goats during early stage (2nd week of infection) and late stage (3rd to 14th week of infection) using immunoblotting assay. Furthermore, no cross reactivity was observed against most commonly found helminths (Trichinella spiralis, Fasciola hepatica, and Toxoplasma gondii) and uninfected goats. The format variables for rHc-CS indirect-ELISA were optimized. The optimum antigen coating concentration was found 0.28μg/well at 37℃ 1h and overnight at 4°C. Optimum dilution ratio of serum and rabbit anti-goat IgG was recorded 1:100 and 1:4000 respectively. The best blocking buffer was 5% bovine serum albumin (BSA) and the best time for blocking, serum incubation and TMB reaction was recorded as 60, 120 and 10 minutes respectively. The cut-off value for positive and negative interpretation was determined as 0.352 (OD450). The diagnostic specificity and sensitivity of the rHc-CS, both were recorded 100%. Conclusion: These results demonstrated that rHc-CS is a potential immunodiagnostic antigen to detect specific antibodies at early and late H. contortus infections in goat.

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Comparison of Fecal Egg Counts and ELISA for the Diagnosis of Dicrocoelium Dendriticum Infection

Cited by 12 publications

References 20 publications

Adhesion-Regulating Molecule from Haemonchus contortus: Potential Antigen for Diagnosis of Early Infection in Goats

Adhesion-Regulating Molecule from Haemonchus contortus: Potential Antigen for Diagnosis of Early Infection in Goats

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

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

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