Recombinant Dense Granule Protein (NcGRA4) Is a Novel Serological Marker for <i>Neospora caninum</i> Infection in Goats
<i>Neospora caninum</i> is widely recognised as one of the most significant causes of abortion in cattle, with infections also occurring in sheep and goats. To prevent and control animal neosporosis, it is crucial to develop sensitive and specific methods for detecting <i>N. caninu...
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| Format: | Article |
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MDPI AG
2023-06-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/13/11/1879 |
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| author | Ruenruetai Udonsom Aongart Mahittikorn Apichai Prachasuphap Kodcharad Jongpitisub Panadda Dhepakson Charoonluk Jirapattharasate |
| author_facet | Ruenruetai Udonsom Aongart Mahittikorn Apichai Prachasuphap Kodcharad Jongpitisub Panadda Dhepakson Charoonluk Jirapattharasate |
| author_sort | Ruenruetai Udonsom |
| collection | DOAJ |
| description | <i>Neospora caninum</i> is widely recognised as one of the most significant causes of abortion in cattle, with infections also occurring in sheep and goats. To prevent and control animal neosporosis, it is crucial to develop sensitive and specific methods for detecting <i>N. caninum</i> infection. Recently, several recombinant proteins have been utilised in serological assays for the diagnosis of neosporosis. In this study, we used commercial gene synthesis to produce dense granular antigen 4 (NcGRA4) recombinant protein. NcGRA4 plasmids were expressed in the <i>Escherichia coli</i> system and then purified. The purified recombinant protein was analysed using sodium dodecyl sulphate–polyacrylamide gel electrophoresis. To evaluate the diagnostic potential of recombinant NcGRA4 protein, we tested 214 serum samples from goat farms via indirect enzyme-linked immunosorbent assay (iELISA) and compared the results to those from the indirect fluorescent antibody test (IFAT). Western blotting analysis revealed a single NcGRA4 band with an expected molecular weight of 32 kDa. The specific IgG against <i>N. caninum</i> was detected in 34.1% and 35% of samples evaluated by NcGRA4 iELISA and IFAT, respectively. The sensitivity and specificity of the NcGRA4 iELISA were 71.6% and 86.3%, respectively, when compared with the results from IFAT. Our results demonstrate that a recombinant protein that can be used to detect animal neosporosis can be produced using a synthetic NcGRA4 gene. Overall, recombinant NcGRA4 shows promise as a sensitive and specific serological marker for identifying target IgG in goat samples. |
| first_indexed | 2024-03-11T03:12:41Z |
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| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-11T03:12:41Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-a3e24aa778e44c1a971cb7983ab3a89a2023-11-18T07:30:55ZengMDPI AGAnimals2076-26152023-06-011311187910.3390/ani13111879Recombinant Dense Granule Protein (NcGRA4) Is a Novel Serological Marker for <i>Neospora caninum</i> Infection in GoatsRuenruetai Udonsom0Aongart Mahittikorn1Apichai Prachasuphap2Kodcharad Jongpitisub3Panadda Dhepakson4Charoonluk Jirapattharasate5Department of Protozoology, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok 10400, ThailandDepartment of Protozoology, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Ratchathewi, Bangkok 10400, ThailandDepartment of Medical Sciences, Medical Life Sciences Institute, 88/7 Tiwanon Road, Talad Kwan Subdistrict, Muang District, Nonthaburi 11000, ThailandDepartment of Medical Sciences, Medical Life Sciences Institute, 88/7 Tiwanon Road, Talad Kwan Subdistrict, Muang District, Nonthaburi 11000, ThailandDepartment of Medical Sciences, Medical Life Sciences Institute, 88/7 Tiwanon Road, Talad Kwan Subdistrict, Muang District, Nonthaburi 11000, ThailandDepartment of Pre-Clinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, 999 Phuthamonthon Sai 4 Rd, Salaya, Nakhon Pathom 73170, Thailand<i>Neospora caninum</i> is widely recognised as one of the most significant causes of abortion in cattle, with infections also occurring in sheep and goats. To prevent and control animal neosporosis, it is crucial to develop sensitive and specific methods for detecting <i>N. caninum</i> infection. Recently, several recombinant proteins have been utilised in serological assays for the diagnosis of neosporosis. In this study, we used commercial gene synthesis to produce dense granular antigen 4 (NcGRA4) recombinant protein. NcGRA4 plasmids were expressed in the <i>Escherichia coli</i> system and then purified. The purified recombinant protein was analysed using sodium dodecyl sulphate–polyacrylamide gel electrophoresis. To evaluate the diagnostic potential of recombinant NcGRA4 protein, we tested 214 serum samples from goat farms via indirect enzyme-linked immunosorbent assay (iELISA) and compared the results to those from the indirect fluorescent antibody test (IFAT). Western blotting analysis revealed a single NcGRA4 band with an expected molecular weight of 32 kDa. The specific IgG against <i>N. caninum</i> was detected in 34.1% and 35% of samples evaluated by NcGRA4 iELISA and IFAT, respectively. The sensitivity and specificity of the NcGRA4 iELISA were 71.6% and 86.3%, respectively, when compared with the results from IFAT. Our results demonstrate that a recombinant protein that can be used to detect animal neosporosis can be produced using a synthetic NcGRA4 gene. Overall, recombinant NcGRA4 shows promise as a sensitive and specific serological marker for identifying target IgG in goat samples.https://www.mdpi.com/2076-2615/13/11/1879recombinant proteindense granulesserology<i>Neospora caninum</i>goats |
| spellingShingle | Ruenruetai Udonsom Aongart Mahittikorn Apichai Prachasuphap Kodcharad Jongpitisub Panadda Dhepakson Charoonluk Jirapattharasate Recombinant Dense Granule Protein (NcGRA4) Is a Novel Serological Marker for <i>Neospora caninum</i> Infection in Goats Animals recombinant protein dense granules serology <i>Neospora caninum</i> goats |
| title | Recombinant Dense Granule Protein (NcGRA4) Is a Novel Serological Marker for <i>Neospora caninum</i> Infection in Goats |
| title_full | Recombinant Dense Granule Protein (NcGRA4) Is a Novel Serological Marker for <i>Neospora caninum</i> Infection in Goats |
| title_fullStr | Recombinant Dense Granule Protein (NcGRA4) Is a Novel Serological Marker for <i>Neospora caninum</i> Infection in Goats |
| title_full_unstemmed | Recombinant Dense Granule Protein (NcGRA4) Is a Novel Serological Marker for <i>Neospora caninum</i> Infection in Goats |
| title_short | Recombinant Dense Granule Protein (NcGRA4) Is a Novel Serological Marker for <i>Neospora caninum</i> Infection in Goats |
| title_sort | recombinant dense granule protein ncgra4 is a novel serological marker for i neospora caninum i infection in goats |
| topic | recombinant protein dense granules serology <i>Neospora caninum</i> goats |
| url | https://www.mdpi.com/2076-2615/13/11/1879 |
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