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    Identification and delineation of members of the Entamoeba complex by pyrosequencing (2010)

    Art
    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Autoren
    Stensvold, C. R.
    Lebbad, M.
    Verweij, J. J.
    Jespersgaard, C.
    Samson-Himmelstjerna, G.
    Nielsen, S. S.
    Henrik, V.
    Quelle
    Molecular and cellular probes : the location, diagnosis and monitoring of disease by specific molecules and cell lines; 24(6) — S. 403–406
    ISSN: 0890-8508
    Sprache
    Englisch
    Verweise
    DOI: 10.1016/j.mcp.2010.07.008
    Pubmed: 20691255
    Kontakt
    Institut für Parasitologie und Tropenveterinärmedizin

    Robert-von-Ostertag-Str. 7-13
    Gebäude 35, 22, 23
    14163 Berlin
    +49 30 838 62310
    parasitologie@vetmed.fu-berlin.de

    Abstract / Zusammenfassung

    A method using a single-round PCR coupled to pyrosequencing was developed for the detection and differentiation of members of the Entamoeba complex. The technique was evaluated using DNA isolated directly from faecal specimens and compared with a duplex real-time PCR targeting Entamoeba histolytica and Entamoeba dispar, and a conventional single-round PCR for the detection of Entamoeba moshkovskii. Tetranucleate cysts from 102 faecal specimens from Swedish, Danish and Dutch patients test-positive for the Entamoeba complex by coproscopic examination were identified to species using each of the three methods. Although none of the patients were confirmed to be positive for E. moshkovskii, E. histolytica and E. dispar were identified in 17 and 86 of the samples, respectively, one of the samples containing both species. There was concordance in results between pyrosequencing and the two other methods used. This study showed that PCR and pyrosequencing could be used for the rapid and high throughput identification of Entamoeba species.

    Copyright © 2010 Elsevier Ltd. All rights reserved.