Molecular identification of <it>Clonorchis sinensis </it>and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA)
<p>Abstract</p> <p>Background</p> <p>Infections with the opisthorchid liver flukes <it>Clonorchis sinensis</it>, <it>Opisthorchis viverrini</it>, and <it>O. felineus </it>cause severe health problems globally, particularly in Southeas...
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| Format: | Article |
| Language: | English |
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BMC
2011-06-01
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| Series: | Parasites & Vectors |
| Online Access: | http://www.parasitesandvectors.com/content/4/1/98 |
| _version_ | 1818786946462253056 |
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| author | Liang Chi Deng Chuanhuan Lv Xiaoli Huang Yan Mao Qiang Liang Pei Xu Jin Sun Jiufeng de Hoog G S Yu Xinbing |
| author_facet | Liang Chi Deng Chuanhuan Lv Xiaoli Huang Yan Mao Qiang Liang Pei Xu Jin Sun Jiufeng de Hoog G S Yu Xinbing |
| author_sort | Liang Chi |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Infections with the opisthorchid liver flukes <it>Clonorchis sinensis</it>, <it>Opisthorchis viverrini</it>, and <it>O. felineus </it>cause severe health problems globally, particularly in Southeast Asia. Early identification of the infection is essential to provide timely and appropriate chemotherapy to patients.</p> <p>Results</p> <p>In this study we evaluate a PCR-based molecular identification method, Multiplex Ligation-dependent Probe Amplification (MLPA), which allows rapid and specific detection of single nucleotide acid differences between <it>Clonorchis sinensis</it>, <it>Opisthorchis viverrini </it>and <it>O. felineus</it>. Three probe pairs were derived from the Internally Transcribed Spacer 1 (ITS1) of three opisthorchid liver flukes using a systematic phylogenetic analysis. Specific loci were detected in all three species, yielding three amplicons with 198,172 and 152 bp, respectively, while no cross reactions were observed. A panel of 66 <it>C. sinensis </it>isolates was screened using MLPA. All species were positively identified, and no inhibition was observed. The detection limit was 10<sup>3 </sup>copies of the ITS gene for the three liver flukes, or about 60 pg genomic DNA for <it>Clonorchis sinensis</it>. Amplification products can be detected by electrophoresis on agarose gel or in a capillary sequencer. In addition, genomic DNA of <it>Clonorchis sinensis </it>in fecal samples of infected rats was positively amplified by MLPA.</p> <p>Conclusion</p> <p>The flexibility and specificity make MLPA a potential tool for specific identification of infections by opisthorchid liver flukes in endemic areas.</p> |
| first_indexed | 2024-12-18T13:59:57Z |
| format | Article |
| id | doaj.art-b9d4903fd71449068d956ebacfeda564 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-12-18T13:59:57Z |
| publishDate | 2011-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-b9d4903fd71449068d956ebacfeda5642022-12-21T21:05:21ZengBMCParasites & Vectors1756-33052011-06-01419810.1186/1756-3305-4-98Molecular identification of <it>Clonorchis sinensis </it>and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA)Liang ChiDeng ChuanhuanLv XiaoliHuang YanMao QiangLiang PeiXu JinSun Jiufengde Hoog G SYu Xinbing<p>Abstract</p> <p>Background</p> <p>Infections with the opisthorchid liver flukes <it>Clonorchis sinensis</it>, <it>Opisthorchis viverrini</it>, and <it>O. felineus </it>cause severe health problems globally, particularly in Southeast Asia. Early identification of the infection is essential to provide timely and appropriate chemotherapy to patients.</p> <p>Results</p> <p>In this study we evaluate a PCR-based molecular identification method, Multiplex Ligation-dependent Probe Amplification (MLPA), which allows rapid and specific detection of single nucleotide acid differences between <it>Clonorchis sinensis</it>, <it>Opisthorchis viverrini </it>and <it>O. felineus</it>. Three probe pairs were derived from the Internally Transcribed Spacer 1 (ITS1) of three opisthorchid liver flukes using a systematic phylogenetic analysis. Specific loci were detected in all three species, yielding three amplicons with 198,172 and 152 bp, respectively, while no cross reactions were observed. A panel of 66 <it>C. sinensis </it>isolates was screened using MLPA. All species were positively identified, and no inhibition was observed. The detection limit was 10<sup>3 </sup>copies of the ITS gene for the three liver flukes, or about 60 pg genomic DNA for <it>Clonorchis sinensis</it>. Amplification products can be detected by electrophoresis on agarose gel or in a capillary sequencer. In addition, genomic DNA of <it>Clonorchis sinensis </it>in fecal samples of infected rats was positively amplified by MLPA.</p> <p>Conclusion</p> <p>The flexibility and specificity make MLPA a potential tool for specific identification of infections by opisthorchid liver flukes in endemic areas.</p>http://www.parasitesandvectors.com/content/4/1/98 |
| spellingShingle | Liang Chi Deng Chuanhuan Lv Xiaoli Huang Yan Mao Qiang Liang Pei Xu Jin Sun Jiufeng de Hoog G S Yu Xinbing Molecular identification of <it>Clonorchis sinensis </it>and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA) Parasites & Vectors |
| title | Molecular identification of <it>Clonorchis sinensis </it>and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA) |
| title_full | Molecular identification of <it>Clonorchis sinensis </it>and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA) |
| title_fullStr | Molecular identification of <it>Clonorchis sinensis </it>and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA) |
| title_full_unstemmed | Molecular identification of <it>Clonorchis sinensis </it>and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA) |
| title_short | Molecular identification of <it>Clonorchis sinensis </it>and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA) |
| title_sort | molecular identification of it clonorchis sinensis it and discrimination with other opisthorchid liver fluke species using multiple ligation depended probe amplification mlpa |
| url | http://www.parasitesandvectors.com/content/4/1/98 |
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