Detection of Toxoplasma gondii oocysts in water sample concentrates by real-time PCR.
PCR techniques in combination with conventional parasite concentration procedures have potential for the sensitive and specific detection of Toxoplasma gondii oocysts in water. Three real-time PCR assays based on the B1 gene and a 529-bp repetitive element were analyzed for the detection of T. gondii tachyzoites and oocysts. Lower sensitivity and specificity were obtained with the B1 gene-based PCR than with the 529-bp repeat-based PCR. New procedures for the real-time PCR detection of T. gondii oocysts in concentrates of surface water were developed and tested in conjunction with a method for the direct extraction of inhibitor-free DNA from water. This technique detected as few as one oocyst seeded to 0.5 ml of packed pellets from water samples concentrated by Envirocheck filters. Thus, this real-time PCR may provide a detection method alternative to the traditional mouse assay and microscopy.
National Center for Zoonotic, Vector-Borne and Enteric Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
Wenli
W
Yang
H D Alan
HD
Lindquist
Vitaliano
V
Cama
Frank W
FW
Schaefer
Eric
E
Villegas
Ronald
R
Fayer
Earl J
EJ
Lewis
Yaoyu
Y
Feng
Lihua
L
Xiao
D000818:Animals; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D033661:Oocysts; D016133:Polymerase Chain Reaction / Q000379:methods*; D015800:Protozoan Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014867:Water / Q000469:parasitology*
D016054:DNA, Protozoan; D015800:Protozoan Proteins; D014867:Water