Next Article in Journal
Effects of Dietary Fiber Sources during Gestation on Stress Status, Abnormal Behaviors and Reproductive Performance of Sows
Previous Article in Journal
Genetic Analysis of Milk Production Traits and Mid-Infrared Spectra in Chinese Holstein Population
Previous Article in Special Issue
Mammary Defences and Immunity against Mastitis in Sheep
Open AccessArticle

Development and Potential Application of Ras Domain Containing Protein from Haemonchus contortus for Diagnosis of Goat Infection

Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
*
Author to whom correspondence should be addressed.
Animals 2020, 10(1), 138; https://doi.org/10.3390/ani10010138
Received: 15 December 2019 / Revised: 5 January 2020 / Accepted: 10 January 2020 / Published: 15 January 2020
(This article belongs to the Special Issue Infectious Diseases in Domestic and Wild Ruminants)
The early diagnosis of Haemonchosis is important for the control and effective treatment of the disease. Early stage detection of Haemonchus contortus infection has not been attempted in goats, even though the immature worms are blood-sucking. This study was carried out to detect H. contortus infection during the early and late stages in goats. The results of this study assessed that recombinant H. contortus Ras domain-containing protein (rHcRas) could detect the antibody in H. contortus-infected goat sera during the prepatent and postpatent periods with high sensitivity and specificity using immunodiagnostic techniques. Our findings illustrated that indirect ELISA based on rHcRas also had the potential to detect H. contortus infection from field samples.
Haemonchus contortus is an important gastrointestinal nematode of small ruminants that causes significant mortality in goats worldwide. Diagnosis of this infection mainly depends on the evaluation of clinical signs and fecal examination. However, limitations often occur in early or mild infections. For this purpose, serological diagnosis seems to be more accurate and reliable. Ras domain-containing protein (Ras) is one of H. contortus’s excretory and secretory products (ESPs) that can be isolated from different larval stages of the nematode. In this study, the recombinant H. contortus Ras domain-containing protein (rHcRas) was expressed and purified and its diagnostic potential was evaluated. Reactions between rHcRas and goat sera were tested using Western blotting (WB). The results showed that rHcRas could be recognized by sera as early as 14 days post infection (DPI), and antibodies against rHcRas in infected goats could be maintained for over 89 days. No reaction was found between rHcRas and antibodies against Trichinella spiralis, Fasciola hepatica, or Toxoplasma gondii. An indirect enzyme-linked immunosorbent assay (ELISA) was produced based on rHcRas. The optimal coating antigen (157 ng of rHcRas/well) and serum dilutions (1:50) were determined via checkerboard titration. Indirect ELISA based on rHcRas showed 87.5% sensitivity and 90.6% specificity. The cut-off values for this experiment were determined to be 0.324 (positive) and 0.273 (negative), respectively, and the variation coefficient (CV) was less than 15%. The results of the indirect ELISA in-field examination showed that 17.6% (9/51) of the goats were infected with H. contortus, higher than the fecal examination results (15.7%, 8/51). When compared the results of the indirect ELISA and necropsy testing, 98.0% (50/51) consistency was found. These results indicated that rHcRas was a potential antigen for the diagnosis of H. contortus infection in goats. View Full-Text
Keywords: Haemonchus contortus; Ras domain-containing protein; serological diagnosis; Western blotting; indirect ELISA Haemonchus contortus; Ras domain-containing protein; serological diagnosis; Western blotting; indirect ELISA
Show Figures

Figure 1

MDPI and ACS Style

Aimulajiang, K.; Cao, M.; Liao, S.; Naqvi, M. .-U.-H.; Tian, X.; Li, Z.; Lu, M.; Lakho, S.A.; Li, X.; Xu, L.; Song, X.; Yan, R. Development and Potential Application of Ras Domain Containing Protein from Haemonchus contortus for Diagnosis of Goat Infection. Animals 2020, 10, 138.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop