Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes
A real-time polymerase chain reaction (PCR) assay with fluorescence resonance energy transfer (FRET) hybridisation probes combined with melting curve analysis was developed to detect Schistosoma japonicum in experimentally infected snails and in faecal samples of infected mice. This procedure is bas...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Fundação Oswaldo Cruz (FIOCRUZ)
2011-11-01
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| Series: | Memorias do Instituto Oswaldo Cruz |
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| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000700008&lng=en&tlng=en |
| _version_ | 1797713391959146496 |
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| author | Tongjit Thanchomnang Pewpan Intapan Pusadee Sri-Aroon Viraphong Lulitanond Penchome Janwan Oranuch Sanpool Wanchai Maleewong |
| author_facet | Tongjit Thanchomnang Pewpan Intapan Pusadee Sri-Aroon Viraphong Lulitanond Penchome Janwan Oranuch Sanpool Wanchai Maleewong |
| author_sort | Tongjit Thanchomnang |
| collection | DOAJ |
| description | A real-time polymerase chain reaction (PCR) assay with fluorescence resonance energy transfer (FRET) hybridisation probes combined with melting curve analysis was developed to detect Schistosoma japonicum in experimentally infected snails and in faecal samples of infected mice. This procedure is based on melting curve analysis of a hybrid between an amplicon from the S. japonicum internal transcribed spacer region 2 sequence, which is a 192-bp S. japonicum-specific sequence, and fluorophore-labelled specific probes. Real-time FRET PCR could detect as little as a single cercaria artificially introduced into a pool of 10 non-infected snails and a single egg inoculated in 100 mg of non-infected mouse faeces. All S. japonicum-infected snails and all faecal samples from infected mice were positive. Non-infected snails, non-infected mouse faeces and genomic DNA from other parasites were negative. This assay is rapid and has potential for epidemiological S. japonicum surveys in snails, intermediate hosts and faecal samples of final hosts. |
| first_indexed | 2024-03-12T07:35:48Z |
| format | Article |
| id | doaj.art-fdaadc88d0e2413c93b10c5311622661 |
| institution | Directory Open Access Journal |
| issn | 1678-8060 |
| language | English |
| last_indexed | 2024-03-12T07:35:48Z |
| publishDate | 2011-11-01 |
| publisher | Fundação Oswaldo Cruz (FIOCRUZ) |
| record_format | Article |
| series | Memorias do Instituto Oswaldo Cruz |
| spelling | doaj.art-fdaadc88d0e2413c93b10c53116226612023-09-02T21:33:11ZengFundação Oswaldo Cruz (FIOCRUZ)Memorias do Instituto Oswaldo Cruz1678-80602011-11-01106783183610.1590/S0074-02762011000700008S0074-02762011000700008Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probesTongjit Thanchomnang0Pewpan Intapan1Pusadee Sri-AroonViraphong Lulitanond2Penchome Janwan3Oranuch Sanpool4Wanchai Maleewong5Mahasarakham UniversityMahidol UniversityKhon Kaen UniversityMahidol UniversityMahidol UniversityMahidol UniversityA real-time polymerase chain reaction (PCR) assay with fluorescence resonance energy transfer (FRET) hybridisation probes combined with melting curve analysis was developed to detect Schistosoma japonicum in experimentally infected snails and in faecal samples of infected mice. This procedure is based on melting curve analysis of a hybrid between an amplicon from the S. japonicum internal transcribed spacer region 2 sequence, which is a 192-bp S. japonicum-specific sequence, and fluorophore-labelled specific probes. Real-time FRET PCR could detect as little as a single cercaria artificially introduced into a pool of 10 non-infected snails and a single egg inoculated in 100 mg of non-infected mouse faeces. All S. japonicum-infected snails and all faecal samples from infected mice were positive. Non-infected snails, non-infected mouse faeces and genomic DNA from other parasites were negative. This assay is rapid and has potential for epidemiological S. japonicum surveys in snails, intermediate hosts and faecal samples of final hosts.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000700008&lng=en&tlng=enSchistosoma japonicumreal-time fluorescence resonance energy transfer PCRDNAsnailsmice |
| spellingShingle | Tongjit Thanchomnang Pewpan Intapan Pusadee Sri-Aroon Viraphong Lulitanond Penchome Janwan Oranuch Sanpool Wanchai Maleewong Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes Memorias do Instituto Oswaldo Cruz Schistosoma japonicum real-time fluorescence resonance energy transfer PCR DNA snails mice |
| title | Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes |
| title_full | Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes |
| title_fullStr | Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes |
| title_full_unstemmed | Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes |
| title_short | Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes |
| title_sort | molecular detection of schistosoma japonicum in infected snails and mouse faeces using a real time pcr assay with fret hybridisation probes |
| topic | Schistosoma japonicum real-time fluorescence resonance energy transfer PCR DNA snails mice |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762011000700008&lng=en&tlng=en |
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