Evaluation of PCR based coprodiagnosis of human opisthorchiasis.
In this study, a recently developed PCR test for the detection of Opisthorchis viverrini in human faecal samples was evaluated using two parasitological methods as references. During a survey of foodborne trematodes (FBT) in the Vientiane Province, Lao PDR, 85 samples were collected and evaluated for FBT eggs by the Kato Katz (KK) technique, the formalin ethyl acetate concentration technique (FECT) and a PCR analysis for the distinction between O. viverrini and other FBT. The two parasitological methods did not differ in the ability of detecting FBT eggs, and a single KK reading was characterized by a sensitivity of 85% when compared to two FECT readings. The PCR tested positive only in cases where eggs had been demonstrated by parasitological examination. However, the PCR tested negative in some samples with very high egg counts. Demonstrating a PCR sensitivity of approximately 50% in samples with faecal egg counts>1000, the previously reported PCR sensitivity based on in vitro studies was not supported. It is believed that technical problems rather than diagnostic reference related issues were responsible for the relatively low PCR performance. Further studies should aim at optimizing DNA extraction and amplification, and future PCR evaluation should include specificity control such as the scanning electron microscopy of eggs in test samples or the expulsion of adult trematodes from PCR tested individuals.
Stensvold
C R
CR
Danish Bilharziasis Laboratory (DBL), Institute for Health Research and Development, Jaergersborg Allé 1D, DK-2920 Charlottenlund, Denmark.
Saijuntha
W
W
Sithithaworn
P
P
Wongratanacheewin
S
S
Strandgaard
H
H
Ornbjerg
N
N
Johansen
M V
MV
D000818:Animals; D018509:DNA, Helminth / Q000032:analysis / Q000302:isolation & purification; D005243:Feces / Q000469:parasitology*; D006801:Humans; D009889:Opisthorchiasis / Q000175:diagnosis* / Q000469:parasitology*; D009891:Opisthorchis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D010270:Parasite Egg Count; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D018509:DNA, Helminth