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Biompha-LAMP: A New Rapid Loop-Mediated Isothermal Amplification Assay for Detecting Schistosoma mansoni in Biomphalaria glabrata Snail Host.
PLoS Negl Trop Dis 2016; 10(12):e0005225PN

Abstract

BACKGROUND

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.

METHODOLOGY/PRINCIPAL FINDINGS

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.

CONCLUSIONS/SIGNIFICANCE

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.

Authors+Show Affiliations

Infectious and Tropical Diseases Group (e-INTRO). IBSAL-CIETUS (Biomedical Research Institute of Salamanca-Research Center for Tropical Diseases at the University of Salamanca), Faculty of Pharmacy, University of Salamanca, Salamanca, Spain.Infectious and Tropical Diseases Group (e-INTRO). IBSAL-CIETUS (Biomedical Research Institute of Salamanca-Research Center for Tropical Diseases at the University of Salamanca), Faculty of Pharmacy, University of Salamanca, Salamanca, Spain.Infectious and Tropical Diseases Group (e-INTRO). IBSAL-CIETUS (Biomedical Research Institute of Salamanca-Research Center for Tropical Diseases at the University of Salamanca), Faculty of Pharmacy, University of Salamanca, Salamanca, Spain.Infectious and Tropical Diseases Group (e-INTRO). IBSAL-CIETUS (Biomedical Research Institute of Salamanca-Research Center for Tropical Diseases at the University of Salamanca), Faculty of Pharmacy, University of Salamanca, Salamanca, Spain.Infectious and Tropical Diseases Group (e-INTRO). IBSAL-CIETUS (Biomedical Research Institute of Salamanca-Research Center for Tropical Diseases at the University of Salamanca), Faculty of Pharmacy, University of Salamanca, Salamanca, Spain.Infectious and Tropical Diseases Group (e-INTRO). IBSAL-CIETUS (Biomedical Research Institute of Salamanca-Research Center for Tropical Diseases at the University of Salamanca), Faculty of Pharmacy, University of Salamanca, Salamanca, Spain.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

27941967

Citation

Gandasegui, Javier, et al. "Biompha-LAMP: a New Rapid Loop-Mediated Isothermal Amplification Assay for Detecting Schistosoma Mansoni in Biomphalaria Glabrata Snail Host." PLoS Neglected Tropical Diseases, vol. 10, no. 12, 2016, pp. e0005225.
Gandasegui J, Fernández-Soto P, Hernández-Goenaga J, et al. Biompha-LAMP: A New Rapid Loop-Mediated Isothermal Amplification Assay for Detecting Schistosoma mansoni in Biomphalaria glabrata Snail Host. PLoS Negl Trop Dis. 2016;10(12):e0005225.
Gandasegui, J., Fernández-Soto, P., Hernández-Goenaga, J., López-Abán, J., Vicente, B., & Muro, A. (2016). Biompha-LAMP: A New Rapid Loop-Mediated Isothermal Amplification Assay for Detecting Schistosoma mansoni in Biomphalaria glabrata Snail Host. PLoS Neglected Tropical Diseases, 10(12), pp. e0005225. doi:10.1371/journal.pntd.0005225.
Gandasegui J, et al. Biompha-LAMP: a New Rapid Loop-Mediated Isothermal Amplification Assay for Detecting Schistosoma Mansoni in Biomphalaria Glabrata Snail Host. PLoS Negl Trop Dis. 2016;10(12):e0005225. PubMed PMID: 27941967.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Biompha-LAMP: A New Rapid Loop-Mediated Isothermal Amplification Assay for Detecting Schistosoma mansoni in Biomphalaria glabrata Snail Host. AU - Gandasegui,Javier, AU - Fernández-Soto,Pedro, AU - Hernández-Goenaga,Juan, AU - López-Abán,Julio, AU - Vicente,Belén, AU - Muro,Antonio, Y1 - 2016/12/12/ PY - 2016/10/05/received PY - 2016/12/01/accepted PY - 2016/12/22/revised PY - 2016/12/13/pubmed PY - 2017/6/14/medline PY - 2016/12/13/entrez SP - e0005225 EP - e0005225 JF - PLoS neglected tropical diseases JO - PLoS Negl Trop Dis VL - 10 IS - 12 N2 - BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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. CONCLUSIONS/SIGNIFICANCE: 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. SN - 1935-2735 UR - https://www.unboundmedicine.com/medline/citation/27941967/Biompha_LAMP:_A_New_Rapid_Loop_Mediated_Isothermal_Amplification_Assay_for_Detecting_Schistosoma_mansoni_in_Biomphalaria_glabrata_Snail_Host_ L2 - http://dx.plos.org/10.1371/journal.pntd.0005225 DB - PRIME DP - Unbound Medicine ER -