Development and Potential Application of Ras Domain Containing Protein from <i>Haemonchus contortus</i> for Diagnosis of Goat Infection
<i>Haemonchus contortus</i> 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 o...
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MDPI AG
2020-01-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/10/1/138 |
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| author | Kalibixiati Aimulajiang Man Cao Shuyi Liao Muhammad Ali-ul-Husnain Naqvi Xiaowei Tian Zehua Li Mingmin Lu Shakeel Ahmed Lakho Xiangrui Li Lixin Xu Xiaokai Song RuoFeng Yan |
| author_facet | Kalibixiati Aimulajiang Man Cao Shuyi Liao Muhammad Ali-ul-Husnain Naqvi Xiaowei Tian Zehua Li Mingmin Lu Shakeel Ahmed Lakho Xiangrui Li Lixin Xu Xiaokai Song RuoFeng Yan |
| author_sort | Kalibixiati Aimulajiang |
| collection | DOAJ |
| description | <i>Haemonchus contortus</i> 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 <i>H. contortus</i>’s excretory and secretory products (ESPs) that can be isolated from different larval stages of the nematode. In this study, the recombinant <i>H. contortus</i> 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 <i>Trichinella spiralis</i>, <i>Fasciola hepatica,</i> or <i>Toxoplasma gondii</i>. 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 <i>H. contortus</i>, 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 <i>H. contortus</i> infection in goats. |
| first_indexed | 2024-12-11T01:30:16Z |
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| last_indexed | 2024-12-11T01:30:16Z |
| publishDate | 2020-01-01 |
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| spelling | doaj.art-dd291b6301934fe591c09845ed4de4492022-12-22T01:25:23ZengMDPI AGAnimals2076-26152020-01-0110113810.3390/ani10010138ani10010138Development and Potential Application of Ras Domain Containing Protein from <i>Haemonchus contortus</i> for Diagnosis of Goat InfectionKalibixiati Aimulajiang0Man Cao1Shuyi Liao2Muhammad Ali-ul-Husnain Naqvi3Xiaowei Tian4Zehua Li5Mingmin Lu6Shakeel Ahmed Lakho7Xiangrui Li8Lixin Xu9Xiaokai Song10RuoFeng Yan11Ministry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMinistry of Education (MOE) Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China<i>Haemonchus contortus</i> 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 <i>H. contortus</i>’s excretory and secretory products (ESPs) that can be isolated from different larval stages of the nematode. In this study, the recombinant <i>H. contortus</i> 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 <i>Trichinella spiralis</i>, <i>Fasciola hepatica,</i> or <i>Toxoplasma gondii</i>. 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 <i>H. contortus</i>, 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 <i>H. contortus</i> infection in goats.https://www.mdpi.com/2076-2615/10/1/138<i>haemonchus contortus</i>ras domain-containing proteinserological diagnosiswestern blottingindirect elisa |
| spellingShingle | Kalibixiati Aimulajiang Man Cao Shuyi Liao Muhammad Ali-ul-Husnain Naqvi Xiaowei Tian Zehua Li Mingmin Lu Shakeel Ahmed Lakho Xiangrui Li Lixin Xu Xiaokai Song RuoFeng Yan Development and Potential Application of Ras Domain Containing Protein from <i>Haemonchus contortus</i> for Diagnosis of Goat Infection Animals <i>haemonchus contortus</i> ras domain-containing protein serological diagnosis western blotting indirect elisa |
| title | Development and Potential Application of Ras Domain Containing Protein from <i>Haemonchus contortus</i> for Diagnosis of Goat Infection |
| title_full | Development and Potential Application of Ras Domain Containing Protein from <i>Haemonchus contortus</i> for Diagnosis of Goat Infection |
| title_fullStr | Development and Potential Application of Ras Domain Containing Protein from <i>Haemonchus contortus</i> for Diagnosis of Goat Infection |
| title_full_unstemmed | Development and Potential Application of Ras Domain Containing Protein from <i>Haemonchus contortus</i> for Diagnosis of Goat Infection |
| title_short | Development and Potential Application of Ras Domain Containing Protein from <i>Haemonchus contortus</i> for Diagnosis of Goat Infection |
| title_sort | development and potential application of ras domain containing protein from i haemonchus contortus i for diagnosis of goat infection |
| topic | <i>haemonchus contortus</i> ras domain-containing protein serological diagnosis western blotting indirect elisa |
| url | https://www.mdpi.com/2076-2615/10/1/138 |
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