Real-time PCR for the detection of Dientamoeba fragilis in fecal samples.
A real-time polymerase chain reaction (PCR) method targeting the 5.8S ribosomal RNA gene was developed for the detection of Dientamoeba fragilis in stool samples. The PCR also included an internal control to detect inhibition of the amplification by fecal constituents in the sample. The assay was performed on species-specific DNA controls (n=29) and a range of stool samples (n=85), and achieved high specificity and sensitivity. D. fragilis DNA could be detected in unpreserved fecal samples up to 8 weeks after storage at 4 degrees C. The use of this assay in a diagnostic laboratory offers the possibility of introducing DNA detection as a feasible technique for the routine diagnosis of intestinal D. fragilis infections.
Verweij
Jaco J
JJ
Leiden University Medical Center, Department of Parasitology, Postbus 9600, 2300 RC Leiden, The Netherlands. j.j.verweij@lumc.nl
Mulder
Bert
B
Poell
Bregje
B
van Middelkoop
Dorien
D
Brienen
Eric A T
EA
van Lieshout
Lisette
L
D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D004029:Dientamoeba / Q000235:genetics* / Q000302:isolation & purification*; D005243:Feces / Q000469:parasitology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*
D016054:DNA, Protozoan