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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 106, No. 7, 2011
Bioline Code: oc11170
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 106, No. 7, 2011

 en Molecular detection of Schistosoma japonicum check for this species in other resources in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes
Thanchomnang, Tongjit; Intapan, Pewpan; Sri-Aroon, Pusadee; Lulitanond, Viraphong; Janwan, Penchome; Sanpool, Oranuch & Maleewong, Wanchai

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 check for this species in other resources 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.

Keywords
Schistosoma japonicum; real-time fluorescence resonance energy transfer PCR; DNA; snails; mice

 
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