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Detection of Entamoeba histolytica by Recombinase Polymerase Amplification

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  • Department of Internal Medicine, University of Texas Medical Branch, Galveston, Texas; Department of Bioengineering, Rice University, Houston, Texas; Facultad de Ciencias Naturales y Matemáticas, Universidad del Rosario, Bogotá, Colombia; Departamento de Medicina Interna, Universidad Nacional de Colombia, Bogotá, Colombia; Departamento de Salud Pública, Universidad Nacional de Colombia, Bogotá, Colombia

Amebiasis is an important cause of diarrheal disease worldwide and has been associated with childhood malnutrition. Traditional microscopy approaches are neither sensitive nor specific for Entamoeba histolytica. Antigen assays are more specific, but many cases are missed unless tested by molecular methods. Although polymerase chain reaction (PCR) is effective, the need for sophisticated, expensive equipment, infrastructure, and trained personnel limits its usefulness, especially in the resource-limited, endemic areas. Here, we report development of a recombinase polymerase amplification (RPA) method to detect E. histolytica specifically. Using visual detection by lateral flow (LF), the test was highly sensitive and specific and could be performed without additional equipment. The availability of this inexpensive, sensitive, and field-applicable diagnostic test could facilitate rapid diagnosis and treatment of amebiasis in endemic regions.

Author Notes

* Address correspondence to Alejandro Castellanos-Gonzalez, Department of Internal Medicine, Division of Infectious Diseases, University of Texas Medical Branch, Galveston, TX 77555. E-mail: alcastel@utmb.edu

Financial support: This study was conducted with the support of the Institute for Translational Sciences at the University of Texas Medical Branch, supported in part by a Clinical and Translational Science Award (UL1TR000071) from the National Center for Advancing Translational Sciences, National Institutes of Health.

Authors' addresses: Gayatri Nair, Mauricio Rebolledo, A. Clinton White Jr., and Alejandro Castellanos-Gonzalez, Department of Internal Medicine, University of Texas Medical Branch, Glaveston, TX, E-mails: gvnair@utmb.edu, mereboll@utmb.edu, acwhite@utmb.edu, and alcastel@utmb.edu. Zachary Crannell and R. Rebecca Richards-Kortum, Department of Bioengineering, Rice University, Houston, TX, E-mails: zcrannell@gmail.com and rkortum@rice.edu. A. Elizabeth Pinilla, Departamento de Medicina Interna, and M. Consuelo López, Departamento de Salud Pública, Universidad Nacional de Colombia, Bogota, Colombia, E-mails: aepinillar@unal.edu.co and mclopezp@unal.edu.co. Juan David Ramírez, Facultad de Ciencias Naturales y Matemáticas, Universidad del Rosario, Grupo de Investigaciones Microbiológicas, Bogota, Colombia, E-mail: juand.ramirez@urosario.edu.co.

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