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Background  A PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed to overcome the need for sensitive techniques for the efficient diagnosis of  Schistosoma  infection in endemic settings with low parasitic burden.    Methodology/Principal Findings  This system amplifies a 121-base pair tandem repeat DNA sequence, immobilizes the resultant 5′ biotinylated product on streptavidin-coated strip-well microplates and uses anti-fluorescein antibodies conjugated to horseradish peroxidase to detect the hybridized fluorescein-labeled oligonucleotide probe. The detection limit of the  Schistosoma  PCR-ELISA system was determined to be 1.3 fg of  S. mansoni  genomic DNA (less than the amount found in a single cell) and estimated to be 0.15  S. mansoni  eggs per gram of feces (fractions of an egg). The system showed good precision and genus specificity since the DNA target was found in seven  Schistosoma  DNA samples:  S. mansoni ,  S. haematobium ,  S. bovis ,  S. intercalatum ,  S. japonicum ,  S. magrebowiei  and  S. rhodaini . By evaluating 206 patients living in an endemic area in Brazil, the prevalence of  S. mansoni  infection was determined to be 18% by examining 12 Kato-Katz slides (41.7 mg/smear, 500 mg total) of a single fecal sample from each person, while the  Schistosoma  PCR-ELISA identified a 30% rate of infection using 500-mg of the same fecal sample. When considering the Kato-Katz method as the reference test, artificial sensitivity and specificity rates of the PCR-ELISA system were 97.4% and 85.1%, respectively. The potential for estimating parasitic load by DNA detection in feces was assessed by comparing absorbance values and eggs per gram of feces, with a Spearman correlation coefficient of 0.700 ( P <0.0001).    Conclusions/Significance  This study reports the development and field evaluation of a sensitive  Schistosoma  PCR-ELISA, a system that may serve as an alternative for diagnosing  Schistosoma  infection.

Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces