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Title: Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes
Authors: Tongjit Thanchomnang
Pewpan Intapan
Pusadee Sri-Aroon
Viraphong Lulitanond
Penchome Janwan
Oranuch Sanpool
Wanchai Maleewong
Khon Kaen University
Mahasarakham University
Mahidol University
Keywords: Medicine
Issue Date: 1-Nov-2011
Citation: Memorias do Instituto Oswaldo Cruz. Vol.106, No.7 (2011), 831-836
Abstract: 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.
ISSN: 16788060
Appears in Collections:Scopus 2011-2015

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