Detection of Toxoplasma gondii in water by an immunomagnetic separation method targeting the sporocysts

An immunomagnetic separation (IMS) method was developed to detect Toxoplasma gondii in fresh waters by using the monoclonal antibody 4B6 targeting the sporocyst wall of T. gondii, Hammondia hammondi, Hammondia heydorni, and Neospora caninum. Water concentrates obtained by filtering 10- to 20-l sampl...

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Bibliographic Details
Published in:Parasitology research (1987) 2007-09, Vol.101 (4), p.989-996
Main Authors: DUMETRE, Aurélien, DARDE, Marie-Laure
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - immunology
Antibodies, Protozoan - immunology
Biological and medical sciences
DNA, Protozoan
Fresh Water - parasitology
Fundamental and applied biological sciences. Psychology
General aspects
General aspects and techniques. Study of several systematic groups. Models
Hammondia hammondi
Hammondia heydorni
Immunomagnetic Separation - methods
Invertebrates
Neospora caninum
Oocysts - immunology
Polymerase Chain Reaction
Rivers - parasitology
Toxoplasma - genetics
Toxoplasma - immunology
Toxoplasma - isolation & purification
Toxoplasma gondii
Water Supply
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Summary:An immunomagnetic separation (IMS) method was developed to detect Toxoplasma gondii in fresh waters by using the monoclonal antibody 4B6 targeting the sporocyst wall of T. gondii, Hammondia hammondi, Hammondia heydorni, and Neospora caninum. Water concentrates obtained by filtering 10- to 20-l samples samples were spiked with Toxoplasma oocysts, sonicated to release the sporocysts, and analyzed by IMS-4B6. Mean sporocyst recoveries were 74.5 ± 5.3% in drinking water, 30.6 ± 2.4 and 37.1 ± 3.2% in surface waters, and 81.6 ± 2.1% in IMS buffer. Then, this IMS method was integrated in a multistep procedure (i.e., filtration, IMS, immunofluorescence and autofluorescence) to detect Toxoplasma in unspiked and spiked water samples (10-30 l) of various qualities. Sporocyst recoveries ranged from 14.4 to 44.7% in drinking water samples spiked with 1-10 oocysts/l, and from 17.8 to 32.5% in surface water samples spiked with 10 oocysts/l. Sporocysts were not detected in 25 unspiked water samples. A sporocyst-like structure was seen in one of these unspiked samples, but its coccidian nature could not be proved by three polymerase chain reaction (PCR) methods targeting sequences of coccidian small and large subunit rRNA genes and Toxoplasma repetitive elements. In conclusion, IMS-4B6 is relevant for the detection of Toxoplasma in water generating small concentrates (
ISSN:0932-0113
1432-1955
DOI:10.1007/s00436-007-0573-0