Application of LAMP Technology in the Molecular Diagnosis of Strongyloides stercoralis Infections
Abstract Strongyloidiasis (Strongyloides stercoralis infection) is a neglected tropical disease of high prevalence worldwide, mainly in tropical and subtropical climates. It ranges from asymptomatic to chronic cases, with relevance in immunocompromised individuals. In severe cases, hyperinfection syndrome and larval dissemination to various organs can occur with high mortality. Diagnosis lacks a standard reference test, which makes it difficult to estimate the accuracy of other diagnostic tests. In previous studies, we developed a molecular method based on LAMP technology for the detection of Strongyloides spp. DNA (Strong-LAMP) with high specificity and sensitivity. Here, we evaluate its performance on patient stool samples collected and stored on filter paper as a simple method of preservation and transport of clinical samples from a Strongyloides endemic area to the laboratory. Twenty samples preserved in filter paper were selected from a total of 192 samples collected and analysed by microscopy and Strong-LAMP in a study conducted in Cubal, Angola. DNA was extracted and analysed in the laboratory. The results indicate that filter paper is a potentially valid carrier for transport and preservation of stool samples for Strong-LAMP molecular diagnosis of strongyloidosis.
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Buonfrate D, Bisanzio D, Giorli G, Odermatt P, Furst T, Greenaway C, et al. The global prevalence of Strongyloides stercoralis infection. Pathogens. 2020;9:468.
Buonfrate D, Formenti F, Perandin F, Bisoffi Z. Novel approaches to the diagnosis of Strongyloides stercoralis infection. Clin Microbiol Infect. 2015; 21(6):543-552.
Buonfrate D, Requena-Mendez A, Angheben A, Cinquini M, Cruciani M, Fittipaldo A, et al. Accuracy of molecular biology techniques for the diagnosis of Strongyloides stercoralis infection—a systematic review and meta-analysis. PloS Negl Trop Dis. 2018;12:e0006229.
Cevallos W, Fernández-Soto P, Calvopiña M, Buendía-Sánchez M, López-Abán J, Vicente B. et al. Diagnosis of amphimeriasis by LAMPhimerus assay in human stool samples long-term storage onto filter paper. PLoS One. 2018; 13(2):e0192637.
Crego-Vicente B, Febrer-Sendra B, Nindia A, Pessela A, Aixut S, Martínez-Campreciós J, Mediavilla A, Silgado A, Sulleiro E, Treviño B, Molina I, Muro A, Salvador F, Fernández-Soto P. First field study using Strong-LAMP for diagnosis of strongyloidiasis in Cubal, Angola. Parasit Vectors. 2023; 31;16(1):393.
Fernández-Soto P, Sánchez-Hernández A, Gandasegui J, Bajo Santos C, López-Abán J, Saugar JM, et al. Strong-LAMP: a LAMP assay for Strongyloides spp. Detection in stool and urine samples towards the diagnosis of human strongyloidiasis starting from a rodent model. PLoS Negl Trop Dis. 2016;10:e0004836.
García-Bernalt Diego J, Fernández-Soto P, Muro A. LAMP in Neglected Tropical Diseases: A Focus on Parasites. Diagnostics. 2021; 11(3):521.
Henríquez-Camacho C, Gotuzzo E, Echevarría J, White AC Jr, Terashima A, Samalvides F et al. Ivermectin versus albendazole or thiabendazole for Strogyloides stercoralis infection. Cochrane Database Syst Rev. 2016; (1):CD007745.
Li Y, Fan P, Zhou S, Zhang L. Loop-mediated isothermal amplification (LAMP): A novel rapid detection platform for pathogens. Microb Pathog. 2017; 107:54-61.
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 2000; 28(12):E63.
Page W, Judd J, Bradbury R. The unique life cycle of Strongyloides stercoralis and implications for public health action. Trop Med Infect Dis. 2018;3:53.
Schär F, Trostdort U, Giardina F, Khieu V, Muth S, Vounatsou P et al. Strongyloides stercoralis: Global Distribution and Risk Factors. Plos Neglect Trop D. 2013; 7(7):e2288.
Soroka M, Wasowicz B, Rymaszewska A. Loop-mediated isothermal amplification (LAMP): the better sibling of PCR? Cells. 2021;10:1931.
Watts MR, James G, Sultana Y, Ginn AN, Outhred AC, Kong F, et al. A loop-mediated isothermal amplification (LAMP) assay for Strongyloides stercoralis in stool that uses a visual detection method with SYTO-82 fluorescent dye. Am J Trop Med Hyg. 2014;90:306-11.
World Health Organization. Soil-transmitted helminth infections. https://www.who.int/news-room/fact-sheets/detail/soil-transmitted-helminth-infections (acceso el 22 de enero de 2023).
Buonfrate D, Formenti F, Perandin F, Bisoffi Z. Novel approaches to the diagnosis of Strongyloides stercoralis infection. Clin Microbiol Infect. 2015; 21(6):543-552.
Buonfrate D, Requena-Mendez A, Angheben A, Cinquini M, Cruciani M, Fittipaldo A, et al. Accuracy of molecular biology techniques for the diagnosis of Strongyloides stercoralis infection—a systematic review and meta-analysis. PloS Negl Trop Dis. 2018;12:e0006229.
Cevallos W, Fernández-Soto P, Calvopiña M, Buendía-Sánchez M, López-Abán J, Vicente B. et al. Diagnosis of amphimeriasis by LAMPhimerus assay in human stool samples long-term storage onto filter paper. PLoS One. 2018; 13(2):e0192637.
Crego-Vicente B, Febrer-Sendra B, Nindia A, Pessela A, Aixut S, Martínez-Campreciós J, Mediavilla A, Silgado A, Sulleiro E, Treviño B, Molina I, Muro A, Salvador F, Fernández-Soto P. First field study using Strong-LAMP for diagnosis of strongyloidiasis in Cubal, Angola. Parasit Vectors. 2023; 31;16(1):393.
Fernández-Soto P, Sánchez-Hernández A, Gandasegui J, Bajo Santos C, López-Abán J, Saugar JM, et al. Strong-LAMP: a LAMP assay for Strongyloides spp. Detection in stool and urine samples towards the diagnosis of human strongyloidiasis starting from a rodent model. PLoS Negl Trop Dis. 2016;10:e0004836.
García-Bernalt Diego J, Fernández-Soto P, Muro A. LAMP in Neglected Tropical Diseases: A Focus on Parasites. Diagnostics. 2021; 11(3):521.
Henríquez-Camacho C, Gotuzzo E, Echevarría J, White AC Jr, Terashima A, Samalvides F et al. Ivermectin versus albendazole or thiabendazole for Strogyloides stercoralis infection. Cochrane Database Syst Rev. 2016; (1):CD007745.
Li Y, Fan P, Zhou S, Zhang L. Loop-mediated isothermal amplification (LAMP): A novel rapid detection platform for pathogens. Microb Pathog. 2017; 107:54-61.
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 2000; 28(12):E63.
Page W, Judd J, Bradbury R. The unique life cycle of Strongyloides stercoralis and implications for public health action. Trop Med Infect Dis. 2018;3:53.
Schär F, Trostdort U, Giardina F, Khieu V, Muth S, Vounatsou P et al. Strongyloides stercoralis: Global Distribution and Risk Factors. Plos Neglect Trop D. 2013; 7(7):e2288.
Soroka M, Wasowicz B, Rymaszewska A. Loop-mediated isothermal amplification (LAMP): the better sibling of PCR? Cells. 2021;10:1931.
Watts MR, James G, Sultana Y, Ginn AN, Outhred AC, Kong F, et al. A loop-mediated isothermal amplification (LAMP) assay for Strongyloides stercoralis in stool that uses a visual detection method with SYTO-82 fluorescent dye. Am J Trop Med Hyg. 2014;90:306-11.
World Health Organization. Soil-transmitted helminth infections. https://www.who.int/news-room/fact-sheets/detail/soil-transmitted-helminth-infections (acceso el 22 de enero de 2023).
Fernández Cencerrado, J., Crego-Vicente, B., Diego del Olmo, M., & Fernández-Soto, P. (2024). Application of LAMP Technology in the Molecular Diagnosis of Strongyloides stercoralis Infections. FarmaJournal, 9(2), 19–29. https://doi.org/10.14201/fj2024921929
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