Detection of schistosomes polymerase chain reaction amplified DNA by oligochromatographic dipstick.
The applications of highly specific and sensitive molecular techniques based on polymerase chain reaction (PCR) have constituted a valuable tool for the diagnosis of schistosomiasis and also for the detection of schistosome infections in the snail intermediate hosts. The common method of detecting PCR amplicons is gel electrophoresis in the presence of ethidium bromide, a carcinogen, which is followed by UV transillumination. Other methods, which are available for detecting PCR products, are real-time PCR, PCR-enzyme-linked immunosorbent assay (PCR-ELIZA) and mass spectrometry but they are cumbersome while they are sometimes complex and expensive. Therefore, a simple method of PCR product detection would be a welcome idea and a most valuable tool particularly in disease endemic countries with limited research facilities and resources. In this study, we applied a simple and rapid method for the detection of Schistosoma haematobium and Schistosoma mansoni PCR amplified DNA products using oligochromatographic (OC) dipstick. The amplicons are visualized by hybridization with a gold conjugated probe, while a control for the chromatographic migration is incorporated in the assay. The lower detection limit observed was 10fg of genomic DNA from each of the two species, while the dipstick was also specific for each of the species used in this study.
Akinwale
O P
OP
Zoology Department, Wolfson Wellcome Biomedical Laboratories, Natural History Museum, London, UK. pheabian@yahoo.co.uk
Laurent
T
T
Mertens
P
P
Leclipteux
T
T
Rollinson
D
D
Kane
R
R
Emery
A
A
Ajayi
M B
MB
Akande
D O
DO
Fesobi
T W
TW
D000818:Animals; D018509:DNA, Helminth / Q000032:analysis* / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000191:economics / Q000379:methods*; D010274:Parasitology / Q000191:economics / Q000379:methods*; D016133:Polymerase Chain Reaction; D012548:Schistosoma haematobium / Q000302:isolation & purification*; D012550:Schistosoma mansoni / Q000302:isolation & purification*; D012552:Schistosomiasis / Q000175:diagnosis*; D012680:Sensitivity and Specificity
D018509:DNA, Helminth