Establishment and preliminary application of PCR-RFLP genotyping method for Giardia duodenalis in goats
Abstract Background Giardia duodenalis (G. duodenalis) is a globally distributed zoonotic protozoan that parasitizes the small intestines of humans and various mammals, such as goats and sheep. The objective of this study was to establish a convenient, accurate, and specific method based on restrict...
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2024-11-01
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| Online Access: | https://doi.org/10.1186/s12917-024-04386-0 |
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| author | Xuanru Mu Jianchao Guo Hongcai Wang Yilong Li Kaijian Yuan Hui Xu Wenjing Zeng Qiaoyu Li Xingang Yu Yang Hong |
| author_facet | Xuanru Mu Jianchao Guo Hongcai Wang Yilong Li Kaijian Yuan Hui Xu Wenjing Zeng Qiaoyu Li Xingang Yu Yang Hong |
| author_sort | Xuanru Mu |
| collection | DOAJ |
| description | Abstract Background Giardia duodenalis (G. duodenalis) is a globally distributed zoonotic protozoan that parasitizes the small intestines of humans and various mammals, such as goats and sheep. The objective of this study was to establish a convenient, accurate, and specific method based on restriction fragment length polymorphism (RFLP) for genotyping assemblages A, B and E of G. duodenalis in goats. The β-giardin gene was amplified using primer pairs bgF1, bgR1, bgF2 and bgR2 by nested PCR. The PCR products were digested with the restriction enzymes Hinf I and Bgl I. The established PCR-RFLP method was used to detect and analyze the genetic subtypes of G. duodenalis in 130 fecal samples from goats and compared simultaneously with microscopic examination and nucleic acid sequencing for G. duodenalis. Results Genetic sequencing confirmed that the PCR-RFLP method accurately distinguished G. duodenalis assemblages A, B and E, as well as different combinations of mixed infections of these three assemblages. Among the 130 samples tested by PCR-RFLP, a total of 26 samples (20.00%) tested positive for G. duodenalis, a higher sensitivity than microscopic examination at 13.85% (18/130). Sequence alignment analysis revealed that among the 26 PCR-positive samples, two were identified as assemblage AI, while the remaining 24 were identified as assemblage E or E12. Conclusions This study established an accurate, efficient and rapid PCR-RFLP genotyping method using the bg sequence of G. duodenalis, enabling accurate identification and effective differentiation of goat-derived G. duodenalis assemblages without requiring sequencing. |
| format | Article |
| id | doaj-art-c95fbe7c5d9c40acb195deb28b7f5e7b |
| institution | DOAJ |
| issn | 1746-6148 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj-art-c95fbe7c5d9c40acb195deb28b7f5e7b2025-08-20T02:49:09ZengBMCBMC Veterinary Research1746-61482024-11-012011910.1186/s12917-024-04386-0Establishment and preliminary application of PCR-RFLP genotyping method for Giardia duodenalis in goatsXuanru Mu0Jianchao Guo1Hongcai Wang2Yilong Li3Kaijian Yuan4Hui Xu5Wenjing Zeng6Qiaoyu Li7Xingang Yu8Yang Hong9School of Animal Science and Technology, Foshan UniversityAgro-Tech Extension Center of Guangdong ProvinceSchool of Animal Science and Technology, Foshan UniversitySchool of Animal Science and Technology, Foshan UniversitySchool of Animal Science and Technology, Foshan UniversitySchool of Animal Science and Technology, Foshan UniversitySchool of Animal Science and Technology, Foshan UniversitySchool of Animal Science and Technology, Foshan UniversitySchool of Animal Science and Technology, Foshan UniversityNational Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, Key Laboratory of Parasite and Vector Biology, Collaborating Center for Tropical Diseases, National Institute of Parasitic Diseases, Chinese Center for Diseases Control and Prevention (Chinese Center for Tropical Diseases Research), National Health Commission of the People’s Republic of China (NHC), World Health Organization (WHO), National Center for International Research on Tropical DiseasesAbstract Background Giardia duodenalis (G. duodenalis) is a globally distributed zoonotic protozoan that parasitizes the small intestines of humans and various mammals, such as goats and sheep. The objective of this study was to establish a convenient, accurate, and specific method based on restriction fragment length polymorphism (RFLP) for genotyping assemblages A, B and E of G. duodenalis in goats. The β-giardin gene was amplified using primer pairs bgF1, bgR1, bgF2 and bgR2 by nested PCR. The PCR products were digested with the restriction enzymes Hinf I and Bgl I. The established PCR-RFLP method was used to detect and analyze the genetic subtypes of G. duodenalis in 130 fecal samples from goats and compared simultaneously with microscopic examination and nucleic acid sequencing for G. duodenalis. Results Genetic sequencing confirmed that the PCR-RFLP method accurately distinguished G. duodenalis assemblages A, B and E, as well as different combinations of mixed infections of these three assemblages. Among the 130 samples tested by PCR-RFLP, a total of 26 samples (20.00%) tested positive for G. duodenalis, a higher sensitivity than microscopic examination at 13.85% (18/130). Sequence alignment analysis revealed that among the 26 PCR-positive samples, two were identified as assemblage AI, while the remaining 24 were identified as assemblage E or E12. Conclusions This study established an accurate, efficient and rapid PCR-RFLP genotyping method using the bg sequence of G. duodenalis, enabling accurate identification and effective differentiation of goat-derived G. duodenalis assemblages without requiring sequencing.https://doi.org/10.1186/s12917-024-04386-0Giardia DuodenalisGenotypePCR-RFLPGoatRestriction enzymes |
| spellingShingle | Xuanru Mu Jianchao Guo Hongcai Wang Yilong Li Kaijian Yuan Hui Xu Wenjing Zeng Qiaoyu Li Xingang Yu Yang Hong Establishment and preliminary application of PCR-RFLP genotyping method for Giardia duodenalis in goats BMC Veterinary Research Giardia Duodenalis Genotype PCR-RFLP Goat Restriction enzymes |
| title | Establishment and preliminary application of PCR-RFLP genotyping method for Giardia duodenalis in goats |
| title_full | Establishment and preliminary application of PCR-RFLP genotyping method for Giardia duodenalis in goats |
| title_fullStr | Establishment and preliminary application of PCR-RFLP genotyping method for Giardia duodenalis in goats |
| title_full_unstemmed | Establishment and preliminary application of PCR-RFLP genotyping method for Giardia duodenalis in goats |
| title_short | Establishment and preliminary application of PCR-RFLP genotyping method for Giardia duodenalis in goats |
| title_sort | establishment and preliminary application of pcr rflp genotyping method for giardia duodenalis in goats |
| topic | Giardia Duodenalis Genotype PCR-RFLP Goat Restriction enzymes |
| url | https://doi.org/10.1186/s12917-024-04386-0 |
| work_keys_str_mv | AT xuanrumu establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT jianchaoguo establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT hongcaiwang establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT yilongli establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT kaijianyuan establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT huixu establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT wenjingzeng establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT qiaoyuli establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT xingangyu establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats AT yanghong establishmentandpreliminaryapplicationofpcrrflpgenotypingmethodforgiardiaduodenalisingoats |