Development of a rapid LATE-PCR-based dipstick for the detection of Schistosomiasis haematobium in human urine samples

Abstract

Background: Schistosomiasis caused by Schistosoma haematobium has become a serious public health problem in tropical and subtropical countries. It is associated with considerable morbidity and mortality in developing and underdeveloped countries. Diagnosis of Schistosoma haematobium infection in both adult and children patients is a clinically important challenge. In the work presented here, we report on an alternative diagnostic method based on presence of the S. haematobium -specific Dra 1, 121 bp repeat fragment in human urine. This study aims to develop Linear-After-The-Exponential- Polymerase Chain Reaction (LATE-PCR) dipstick for the detection of Schistosoma haematobium in urine samples.

Methodology: In an effort to developed or improved the current PCR techniques LATE-PCR Lateral flow was developed, in order to allow specie-specific amplification of Schistosoma haematobium in urine samples.

Results: Evaluation study on 50 urine samples showed 100% sensitivity and specificity suggesting the primers and designed probe is specific. Sensitivity of LATE-PCR currently was said to be higher when compared to parasitological methods (Microscopy). LATE-PCR dipstick is highly sensitive since 30 positive urine samples with very low eggs burden were also seen to be positive by the test. The analytical sensitivity was performed and detection limit determine for Schistosoma haematobium. The detection limit of the PCR reaction on Schistosoma haematobium was found to be 1fg/ ?l using 10 fold serial dilution.

Conclusion: LATE-PCR dipstick developed in this study provides a valuable alternative for the detection of Schistosoma haematobium infection in the study area to speed up diagnosis in addition to the conventional method currently used.

Keywords: Polymerase chain reaction, Lateral flow assay, Schistosoma haematobium, Urine samples