Development of an immunomagnetic separation-polymerase chain reaction (IMS-PCR) assay specific for Enterocytozoon bieneusi in water samples.
OBJECTIVE
Microsporidia have become widely recognized as important human pathogens. Among Microsporidia, Enterocytozoon bieneusi is responsible for severe gastrointestinal disease. To date, no current therapy has been proven effective. Their mode of transmission and environmental occurrence are poorly documented because of the lack of detection methods that are both species-specific and sensitive. In this study, we developed a sensitive and specific molecular method to detect E. bieneusi spores in water samples.


RESULTS
The molecular assay combined immunomagnetic separation (IMS) and polymerase chain reaction (PCR) amplification to detect E. bieneusi spores. A comparison was made of IMS magnetic beads coated with two different monoclonal antibodies, one specific for the Encephalitozoon genus that cross-reacts with E. bieneusi and the other specific only for the E. bieneusi species itself.


CONCLUSIONS
Immunotech beads coated with the antibody specific for E. bieneusi were found to be the most effective combination.


CONCLUSIONS
The highly specific IMS-PCR assay developed in this study provides a rapid and sensitive means of screening water samples for the presence of E. bieneusi spores.
Sorel
N
N
Laboratoire de parasitologie, CHU de Nantes, 9 quai Moncousu, Nantes cedex 1, France.
Guillot
E
E
Thellier
M
M
Accoceberry
I
I
Datry
A
A
Mesnard-Rouiller
L
L
Miégeville
M
M
D000913:Antibodies, Protozoan / Q000032:analysis; D016054:DNA, Protozoan / Q000032:analysis; D021862:Enterocytozoon / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D016881:Microsporidiosis / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014871:Water Microbiology
D000913:Antibodies, Protozoan; D016054:DNA, Protozoan