title: Molecular detection of Schistosoma japonicum in infected snails and mouse faeces using a real-time PCR assay with FRET hybridisation probes.
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.
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, 40002 Khon Kaen, Thailand.
authors: Tongjit
authors: T
authors: Thanchomnang
authors: Pewpan
authors: P
authors: Intapan
authors: Pusadee
authors: P
authors: Sri-Aroon
authors: Viraphong
authors: V
authors: Lulitanond
authors: Penchome
authors: P
authors: Janwan
authors: Oranuch
authors: O
authors: Sanpool
authors: Wanchai
authors: W
authors: Maleewong
mesh_terms: D000818:Animals; D018509:DNA, Helminth / Q000032:analysis*; D005243:Feces / Q000469:parasitology*; D031541:Fluorescence Resonance Energy Transfer; D051379:Mice; D009693:Nucleic Acid Hybridization; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012549:Schistosoma japonicum / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012908:Snails / Q000469:parasitology*
chemical_list: D018509:DNA, Helminth