title: Development of multiplex real-time polymerase chain reaction for detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in clinical specimens.
abstract: Multiplex real-time polymerase chain reaction (PCR) was developed for differential detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii. Specific primers were designed for all three species, and then differentiation of E. histolytica and E. dispar was achieved simultaneously using a hybridization probe and melting curve analysis, whereas E. moshkovskii was detected with a separate probe under the same condition. This assay detected as little as 0.2 pg of E. histolytica DNA and 2 pg each for E. dispar and E. moshkovskii DNA. Thirty-five clinical samples suspected to be E. histolytica infection by microscopy were tested. The results showed 32 positive samples; four samples were E. histolytica and 28 samples were E. dispar. Interestingly, one E. dispar positive sample showed a mixed infection with E. moshkovskii. This is the first report of E. moshkovskii infection from Thailand and this assay is currently the most rapid and sensitive method to differentiate these human amoebas.
authors.affiliation: Department of Protozoology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. zulhainan@hotmail.com
authors.forename: Zulhainan
authors.initials: Z
authors.lastname: Hamzah
authors.forename: Songsak
authors.initials: S
authors.lastname: Petmitr
authors.forename: Mathirut
authors.initials: M
authors.lastname: Mungthin
authors.forename: Saovanee
authors.initials: S
authors.lastname: Leelayoova
authors.forename: Porntip
authors.initials: P
authors.lastname: Chavalitshewinkoon-Petmitr
mesh_terms: D016054:DNA, Protozoan / Q000032:analysis; D004747:Entamoeba / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004749:Entamoebiasis / Q000175:diagnosis* / Q000453:epidemiology / Q000469:parasitology; D005243:Feces / Q000469:parasitology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013785:Thailand / Q000453:epidemiology; D013997:Time Factors
chemical_list: D016054:DNA, Protozoan