A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections
Abstract Background Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are,...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-10-01
|
| Series: | BMC Infectious Diseases |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12879-019-4512-3 |
| _version_ | 1818126427097137152 |
|---|---|
| author | Guo-Qiang Zhu Li Li John Asekhaen Ohiolei Yan-Tao Wu Wen-Hui Li Nian-Zhang Zhang Bao-Quan Fu Hong-Bin Yan Wan-Zhong Jia |
| author_facet | Guo-Qiang Zhu Li Li John Asekhaen Ohiolei Yan-Tao Wu Wen-Hui Li Nian-Zhang Zhang Bao-Quan Fu Hong-Bin Yan Wan-Zhong Jia |
| author_sort | Guo-Qiang Zhu |
| collection | DOAJ |
| description | Abstract Background Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are, so far, no commercially available molecular diagnostic kits capable of simultaneously detecting all four parasites in dogs. The aim of the study was therefore to develop a multiplex PCR assay that will accurately detect all four cestode infections in one reaction. Methods Specific primers for a multiplex PCR were designed based on corresponding mitochondrial genome sequences, and its detection limit was assessed by serial dilutions of the genomic DNAs of tapeworms examined. Furthermore, field samples of dog feces were tested using the developed assay. Results A multiplex polymerase chain reaction (PCR) assay was developed based on mitochondrial DNA (mtDNA) that accurately and simultaneously identify four cestode species in one reaction using specific fragment sizes of 592, 385, 283, and 190 bp for T. hydatigena, T. multiceps, T. pisiformis, and D. caninum, respectively. The lowest DNA concentration detected was 1 ng for T. hydatigena, T. multiceps and T. pisiformis, and 0.1 ng for D. caninum in a 25 μl reaction system. This assay offers high potential for the rapid detection of these four tapeworms in host feces simultaneously. Conclusions This study provides an efficient tool for the simultaneous detection of T. hydatigena, T. multiceps, T. pisiformis, and D. caninum. The assay will be potentially useful in epidemiological studies, diagnosis, and treatment of these four cestodes infections during prevention and control program. |
| first_indexed | 2024-12-11T07:01:17Z |
| format | Article |
| id | doaj.art-66b7b2f5537b42eba4dd9e3e90f6c4ad |
| institution | Directory Open Access Journal |
| issn | 1471-2334 |
| language | English |
| last_indexed | 2024-12-11T07:01:17Z |
| publishDate | 2019-10-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Infectious Diseases |
| spelling | doaj.art-66b7b2f5537b42eba4dd9e3e90f6c4ad2022-12-22T01:16:37ZengBMCBMC Infectious Diseases1471-23342019-10-011911710.1186/s12879-019-4512-3A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infectionsGuo-Qiang Zhu0Li Li1John Asekhaen Ohiolei2Yan-Tao Wu3Wen-Hui Li4Nian-Zhang Zhang5Bao-Quan Fu6Hong-Bin Yan7Wan-Zhong Jia8State Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASState Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASState Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASState Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASState Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASState Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASState Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASState Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASState Key Laboratory of Veterinary Etiological Biology/ Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute, CAASAbstract Background Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum are four common large and medium-sized tapeworms parasitizing the small intestine of dogs and other canids. These parasites cause serious impact on the health and development of livestock. However, there are, so far, no commercially available molecular diagnostic kits capable of simultaneously detecting all four parasites in dogs. The aim of the study was therefore to develop a multiplex PCR assay that will accurately detect all four cestode infections in one reaction. Methods Specific primers for a multiplex PCR were designed based on corresponding mitochondrial genome sequences, and its detection limit was assessed by serial dilutions of the genomic DNAs of tapeworms examined. Furthermore, field samples of dog feces were tested using the developed assay. Results A multiplex polymerase chain reaction (PCR) assay was developed based on mitochondrial DNA (mtDNA) that accurately and simultaneously identify four cestode species in one reaction using specific fragment sizes of 592, 385, 283, and 190 bp for T. hydatigena, T. multiceps, T. pisiformis, and D. caninum, respectively. The lowest DNA concentration detected was 1 ng for T. hydatigena, T. multiceps and T. pisiformis, and 0.1 ng for D. caninum in a 25 μl reaction system. This assay offers high potential for the rapid detection of these four tapeworms in host feces simultaneously. Conclusions This study provides an efficient tool for the simultaneous detection of T. hydatigena, T. multiceps, T. pisiformis, and D. caninum. The assay will be potentially useful in epidemiological studies, diagnosis, and treatment of these four cestodes infections during prevention and control program.http://link.springer.com/article/10.1186/s12879-019-4512-3Multiplex PCRCanidsTapewormsMitochondrial DNA |
| spellingShingle | Guo-Qiang Zhu Li Li John Asekhaen Ohiolei Yan-Tao Wu Wen-Hui Li Nian-Zhang Zhang Bao-Quan Fu Hong-Bin Yan Wan-Zhong Jia A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections BMC Infectious Diseases Multiplex PCR Canids Tapeworms Mitochondrial DNA |
| title | A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections |
| title_full | A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections |
| title_fullStr | A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections |
| title_full_unstemmed | A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections |
| title_short | A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis, and Dipylidium caninum infections |
| title_sort | multiplex pcr assay for the simultaneous detection of taenia hydatigena t multiceps t pisiformis and dipylidium caninum infections |
| topic | Multiplex PCR Canids Tapeworms Mitochondrial DNA |
| url | http://link.springer.com/article/10.1186/s12879-019-4512-3 |
| work_keys_str_mv | AT guoqiangzhu amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT lili amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT johnasekhaenohiolei amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT yantaowu amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT wenhuili amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT nianzhangzhang amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT baoquanfu amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT hongbinyan amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT wanzhongjia amultiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT guoqiangzhu multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT lili multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT johnasekhaenohiolei multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT yantaowu multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT wenhuili multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT nianzhangzhang multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT baoquanfu multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT hongbinyan multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections AT wanzhongjia multiplexpcrassayforthesimultaneousdetectionoftaeniahydatigenatmulticepstpisiformisanddipylidiumcaninuminfections |