Evaluation of loop-mediated isothermal amplification suitable for molecular monitoring of schistosome-infected snails in field laboratories

• Published in: The American journal of tropical medicine and hygiene 2013-02, Vol.88 (2), p.344-351
• Main Authors: Hamburger, Joseph, Abbasi, Ibrahim, Kariuki, Curtis, Wanjala, Atsabina, Mzungu, Elton, Mungai, Peter, Muchiri, Eric, King, Charles H
• Format: Article
• Language: English
• Subjects: Animals; DNA, Helminth - genetics; DNA, Helminth - isolation & purification; Environmental Monitoring - methods; Polymerase Chain Reaction - methods; Schistosoma haematobium; Schistosoma mansoni; Schistosoma mansoni - genetics; Schistosoma mansoni - isolation & purification; Sensitivity and Specificity; Sequence Analysis, DNA; Snails - parasitology
• ISSN: 0002-9637; 1476-1645
• DOI: 10.4269/ajtmh.2012.12-0208

We previously described loop-mediated isothermal amplification (LAMP) for detection of Schistosoma haematobium and S. mansoni DNA in infected snails. In the present study, we adapted the LAMP assay for application in field laboratories in schistosomiasis-endemic areas. Isolation of DNA was simplified by blotting snail tissue (extracted in NaOH/sodium dodecyl sulfate) onto treated membranes, which enabled preservation at ambient temperatures. A ready-mix of LAMP reagents, suitable for shipment at ambient temperature and storage in minimal refrigeration, was used. Local survey teams without experience in molecular biology acquired operational expertise with this test within a few hours. Fifty-four field-caught snails were tested locally by LAMP and 59 were tested at similar conditions in Jerusalem. The LAMP results were consistent with those of a polymerase chain reaction; only four samples showed false-negative results. Results indicate that LAMP assays are suitable for detection of S. haematobium and S. mansoni in low-technology parasitology laboratories in which schistosomiasis elimination activities are undertaken.