Adhesion-Regulating Molecule from <i>Haemonchus contortus</i>: Potential Antigen for Diagnosis of Early Infection in Goats
<i>Haemonchus contortus</i>, 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...
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MDPI AG
2019-12-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/9/1/34 |
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| author | Kalibixiati Aimulajiang Muhammad Ali-ul-Husnain Naqvi Wen Chu Mingmin Lu Xiaowei Tian Yongqian Bu Muhammad Ali Memon Xiangrui Li Lixin Xu Xiaokai Song Ruofeng Yan |
| author_facet | Kalibixiati Aimulajiang Muhammad Ali-ul-Husnain Naqvi Wen Chu Mingmin Lu Xiaowei Tian Yongqian Bu Muhammad Ali Memon Xiangrui Li Lixin Xu Xiaokai Song Ruofeng Yan |
| author_sort | Kalibixiati Aimulajiang |
| collection | DOAJ |
| description | <i>Haemonchus contortus</i>, 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 <i>H. contortus</i> adhesion-regulating molecule protein (rHCADRM) was expressed and purified, and its diagnostic potential was evaluated. Serum samples from goats experimentally infected with <i>H. contortus</i> (<i>n</i> = 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 <i>Trichinella spiralis</i>, <i>Fasciola hepatica</i>, or <i>Toxoplasma gondii</i>. 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 <i>H. contortus</i> 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 <i>H. contortus</i>, 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 <i>H. contortus</i> infection in goats. |
| first_indexed | 2024-04-13T00:25:27Z |
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| last_indexed | 2024-04-13T00:25:27Z |
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| spelling | doaj.art-319734fb08ed40b5ab9df65a9dde91892022-12-22T03:10:37ZengMDPI AGPathogens2076-08172019-12-01913410.3390/pathogens9010034pathogens9010034Adhesion-Regulating Molecule from <i>Haemonchus contortus</i>: Potential Antigen for Diagnosis of Early Infection in GoatsKalibixiati Aimulajiang0Muhammad Ali-ul-Husnain Naqvi1Wen Chu2Mingmin Lu3Xiaowei Tian4Yongqian Bu5Muhammad Ali Memon6Xiangrui Li7Lixin Xu8Xiaokai Song9Ruofeng Yan10MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China<i>Haemonchus contortus</i>, 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 <i>H. contortus</i> adhesion-regulating molecule protein (rHCADRM) was expressed and purified, and its diagnostic potential was evaluated. Serum samples from goats experimentally infected with <i>H. contortus</i> (<i>n</i> = 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 <i>Trichinella spiralis</i>, <i>Fasciola hepatica</i>, or <i>Toxoplasma gondii</i>. 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 <i>H. contortus</i> 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 <i>H. contortus</i>, 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 <i>H. contortus</i> infection in goats.https://www.mdpi.com/2076-0817/9/1/34<i>haemonchus contortus</i>adhesion regulating moleculeserological diagnosiswestern blottingindirect elisa |
| spellingShingle | Kalibixiati Aimulajiang Muhammad Ali-ul-Husnain Naqvi Wen Chu Mingmin Lu Xiaowei Tian Yongqian Bu Muhammad Ali Memon Xiangrui Li Lixin Xu Xiaokai Song Ruofeng Yan Adhesion-Regulating Molecule from <i>Haemonchus contortus</i>: Potential Antigen for Diagnosis of Early Infection in Goats Pathogens <i>haemonchus contortus</i> adhesion regulating molecule serological diagnosis western blotting indirect elisa |
| title | Adhesion-Regulating Molecule from <i>Haemonchus contortus</i>: Potential Antigen for Diagnosis of Early Infection in Goats |
| title_full | Adhesion-Regulating Molecule from <i>Haemonchus contortus</i>: Potential Antigen for Diagnosis of Early Infection in Goats |
| title_fullStr | Adhesion-Regulating Molecule from <i>Haemonchus contortus</i>: Potential Antigen for Diagnosis of Early Infection in Goats |
| title_full_unstemmed | Adhesion-Regulating Molecule from <i>Haemonchus contortus</i>: Potential Antigen for Diagnosis of Early Infection in Goats |
| title_short | Adhesion-Regulating Molecule from <i>Haemonchus contortus</i>: Potential Antigen for Diagnosis of Early Infection in Goats |
| title_sort | adhesion regulating molecule from i haemonchus contortus i potential antigen for diagnosis of early infection in goats |
| topic | <i>haemonchus contortus</i> adhesion regulating molecule serological diagnosis western blotting indirect elisa |
| url | https://www.mdpi.com/2076-0817/9/1/34 |
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