Biompha-LAMP: A New Rapid Loop-Mediated Isothermal Amplification Assay for Detecting Schistosoma mansoni in Biomphalaria glabrata Snail Host

• Published in: PLoS neglected tropical diseases 2016-12, Vol.10 (12), p.e0005225
• Main Authors: Gandasegui, Javier, Fernández-Soto, Pedro, Hernández-Goenaga, Juan, López-Abán, Julio, Vicente, Belén, Muro, Antonio
• Format: Article
• Language: English
• Subjects: Analysis; Animals; Biology and Life Sciences; Biomedical research; Biomphalaria - parasitology; Cell division; Deoxyribonucleic acid; DNA; DNA Primers; DNA, Helminth - analysis; DNA, Helminth - isolation & purification; Funding; Human blood fluke; Infections; Laboratories; Medical research; Medicine and Health Sciences; Mollusks; Nucleic Acid Amplification Techniques - methods; Parasites; Parasitic diseases; Pharmacy; Polymerase Chain Reaction; Pulmonates; Research and analysis methods; Schistosoma mansoni - genetics; Schistosoma mansoni - isolation & purification; Schistosomiasis; Sodium hydroxide; Studies; Tropical diseases
• ISSN: 1935-2735; 1935-2727; 1935-2735
• DOI: 10.1371/journal.pntd.0005225

Schistosomiasis remains one of the most common endemic parasitic diseases affecting over 230 million people worlwide. Schistosoma mansoni is the main species causing intestinal and hepatic schistosomiasis and the fresh water pulmonate snails of the genus Biomphalaria are best known for their role as intermediate hosts of the parasite. The development of new molecular monitoring assays for large-scale screening of snails from transmission sites to detect the presence of schistosomes is an important point to consider for snail control interventions related to schistosomiasis elimination. Our work was focussed on developing and evaluating a new LAMP assay combined with a simple DNA extraction method to detect S. mansoni in experimentally infected snails as a diagnostic tool for field conditions. A LAMP assay using a set of six primers targeting a sequence of S. mansoni ribosomal intergenic spacer 28S-18S rRNA was designed. The detection limit of the LAMP assay was 0.1 fg of S. mansoni DNA at 63°C for 50 minutes. LAMP was evaluated by examining S. mansoni DNA in B. glabrata snails experimentally exposed to miracidia at different times post-exposure: early prepatent period (before cercarial shedding), light infections (snails exposed to a low number of miracidia) and detection of infected snails in pooled samples (within a group of uninfected snails). DNA for LAMP assays was obtained by using a commercial DNA extraction kit or a simple heat NaOH extraction method. We detected S. mansoni DNA in all groups of snails by using no complicated requirement procedure for DNA obtaining. Our LAMP assay, named Biompha-LAMP, is specific, sensitive, rapid and potentially adaptable as a cost-effective method for screening of intermediate hosts infected with S. mansoni in both individual snails and pooled samples. The assay could be suitable for large-scale field surveys for schistosomes control campaigns in endemic areas.