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A tool for diagnosis of Dicrocoelium dendriticum infection: hatching eggs and molecular identification of the miracidium.
Parasitol Res. 2013 Apr; 112(4):1589-95.PR

Abstract

DNA primers were designed from the 18S rRNA sequence from the relevant digenean trematode Dicrocoelium dendriticum to evaluate a polymerase chain reaction-based diagnostic method of this parasite from its eggs in faeces of naturally and experimentally infected sheep. In order to get DNA from D. dendriticum eggs, several hatching mechanisms were studied. Successful results were obtained when the eggs were frozen to -80 °C and/or in liquid nitrogen and then defrosted. This method allowed the opening of the egg operculum and the liberation of the miracidium. DNA from D. dendriticum adults and from hatching egg miracidia was obtained and an amplification single band of 1.95 kb was observed using primers designed for the total 18S rRNA sequence in both cases as well as when the template DNA was from adults of the closely related parasite Fasciola hepatica; in addition, a single and specific 0.8-kb band was obtained when primers based on an internal partial 18S rRNA sequence were used. The method showed to be useful not only in samples coming from adults, but in eggs from gall bladder and faeces as well. F. hepatica internal 18S rRNA primers were also designed and used as a negative control to prove that the eggs in faeces came from D. dendriticum and not from F. hepatica. A molecular tool able to detect a minimum of about 40 D. dendriticum eggs in one of the definitive host faeces has been developed for the first time and could provide a useful molecular tool to improve the conventional coprological diagnosis for detecting D. dendriticum eggs.

Authors+Show Affiliations

Departamento de Sanidad Animal, Instituto de Ganadería de Montaña, Consejo Superior de Investigaciones Científicas (CSIC)- Universidad de León (ULE), Grulleros, 24346, León, Spain. mhsanh@unileon.esNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23385970

Citation

Sandoval, H, et al. "A Tool for Diagnosis of Dicrocoelium Dendriticum Infection: Hatching Eggs and Molecular Identification of the Miracidium." Parasitology Research, vol. 112, no. 4, 2013, pp. 1589-95.
Sandoval H, Manga-González MY, Castro JM. A tool for diagnosis of Dicrocoelium dendriticum infection: hatching eggs and molecular identification of the miracidium. Parasitol Res. 2013;112(4):1589-95.
Sandoval, H., Manga-González, M. Y., & Castro, J. M. (2013). A tool for diagnosis of Dicrocoelium dendriticum infection: hatching eggs and molecular identification of the miracidium. Parasitology Research, 112(4), 1589-95. https://doi.org/10.1007/s00436-013-3313-7
Sandoval H, Manga-González MY, Castro JM. A Tool for Diagnosis of Dicrocoelium Dendriticum Infection: Hatching Eggs and Molecular Identification of the Miracidium. Parasitol Res. 2013;112(4):1589-95. PubMed PMID: 23385970.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A tool for diagnosis of Dicrocoelium dendriticum infection: hatching eggs and molecular identification of the miracidium. AU - Sandoval,H, AU - Manga-González,M Yolanda, AU - Castro,José M, Y1 - 2013/02/06/ PY - 2012/12/13/received PY - 2013/01/18/accepted PY - 2013/2/7/entrez PY - 2013/2/7/pubmed PY - 2013/8/29/medline SP - 1589 EP - 95 JF - Parasitology research JO - Parasitol. Res. VL - 112 IS - 4 N2 - DNA primers were designed from the 18S rRNA sequence from the relevant digenean trematode Dicrocoelium dendriticum to evaluate a polymerase chain reaction-based diagnostic method of this parasite from its eggs in faeces of naturally and experimentally infected sheep. In order to get DNA from D. dendriticum eggs, several hatching mechanisms were studied. Successful results were obtained when the eggs were frozen to -80 °C and/or in liquid nitrogen and then defrosted. This method allowed the opening of the egg operculum and the liberation of the miracidium. DNA from D. dendriticum adults and from hatching egg miracidia was obtained and an amplification single band of 1.95 kb was observed using primers designed for the total 18S rRNA sequence in both cases as well as when the template DNA was from adults of the closely related parasite Fasciola hepatica; in addition, a single and specific 0.8-kb band was obtained when primers based on an internal partial 18S rRNA sequence were used. The method showed to be useful not only in samples coming from adults, but in eggs from gall bladder and faeces as well. F. hepatica internal 18S rRNA primers were also designed and used as a negative control to prove that the eggs in faeces came from D. dendriticum and not from F. hepatica. A molecular tool able to detect a minimum of about 40 D. dendriticum eggs in one of the definitive host faeces has been developed for the first time and could provide a useful molecular tool to improve the conventional coprological diagnosis for detecting D. dendriticum eggs. SN - 1432-1955 UR - https://www.unboundmedicine.com/medline/citation/23385970/A_tool_for_diagnosis_of_Dicrocoelium_dendriticum_infection:_hatching_eggs_and_molecular_identification_of_the_miracidium_ L2 - https://dx.doi.org/10.1007/s00436-013-3313-7 DB - PRIME DP - Unbound Medicine ER -