Development of a real-time polymerase-chain-reaction assay for quantitative detection of Enterocytozoon bieneusi DNA in stool specimens from immunocompromised patients with intestinal microsporidiosis.
A new real-time polymerase chain reaction (PCR) method was developed for quantitation of Enterocytozoon bieneusi DNA in sequential stool specimens from immunocompromised patients with intestinal microsporidiosis. Patients were treated with fumagillin (n=6) or with placebo (n=6), in a randomized comparative trial. At baseline, mean E. bieneusi DNA levels were not significantly different in stool specimens from the placebo group, compared with those from the fumagillin group (5.9+/-0.4 vs. 5.9+/-0.6 log(10) copies/microL of stool suspension, respectively; P=.96). In the placebo group, parasitic burden remained stable during follow-up (P=.46), whereas, in the fumagillin group, E. bieneusi DNA levels dropped below the lower limit of detection in all patients (mean reduction from baseline, -4.7 log(10) copies; P<.0001). Real-time PCR performed better than did semiquantitative assessments by microscopy, to measure parasitic burden. In conclusion, this real-time PCR assay is a reliable tool for quantitation of E. bieneusi DNA in stool specimens and for the monitoring of treatment efficacy.
Menotti
Jean
J
Laboratory of Parasitology-Mycology, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris, and Faculté de Médecine Lariboisière-Saint-Louis, Université Paris VII, Paris, France.
Cassinat
Bruno
B
Porcher
Raphaël
R
Sarfati
Claudine
C
Derouin
Francis
F
Molina
Jean-Michel
JM
D000981:Antiprotozoal Agents / Q000627:therapeutic use; D003510:Cyclohexanes; D016054:DNA, Protozoan / Q000032:analysis* / Q000235:genetics; D021862:Enterocytozoon / Q000235:genetics* / Q000302:isolation & purification*; D005231:Fatty Acids, Unsaturated / Q000627:therapeutic use; D005243:Feces / Q000469:parasitology*; D006801:Humans; D016867:Immunocompromised Host; D007411:Intestinal Diseases, Parasitic / Q000175:diagnosis / Q000188:drug therapy / Q000469:parasitology; D016881:Microsporidiosis / Q000175:diagnosis* / Q000188:drug therapy / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012717:Sesquiterpenes
D000981:Antiprotozoal Agents; D003510:Cyclohexanes; D016054:DNA, Protozoan; D005231:Fatty Acids, Unsaturated; D012717:Sesquiterpenes; C026211:fumagillin