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-
1.
Lozano R, Naghavi M, Foreman K, Lim S, Shibuya K, Aboyans V, Abraham J, Adair T, Aggarwal R, Ahn SY, Alvarado M, Anderson HR, Anderson LM, Andrews KG, Atkinson C, Baddour LM, Barker-Collo S, Bartels DH, Bell ML, Benjamin EJ, Bennett D, Bhalla K, Bikbov B, Bin Abdulhak A, Birbeck G, Blyth F, Bolliger I, Boufous S, Bucello C, Burch M, Burney P, Carapetis J, Chen H, Chou D, Chugh SS, Coffeng LE, Colan SD, Colquhoun S, Colson KE, Condon J, Connor MD, Cooper LT, Corriere M, Cortinovis M, de Vaccaro KC, Couser W, Cowie BC, Criqui MH, Cross M, Dabhadkar KC, Dahodwala N, De Leo D, Degenhardt L, Delossantos A, Denenberg J, Des Jarlais DC, Dharmaratne SD, Dorsey ER, Driscoll T, Duber H, Ebel B, Erwin PJ, Espindola P, Ezzati M, Feigin V, Flaxman AD, Forouzanfar MH, Fowkes FG, Franklin R, Fransen M, Freeman MK, Gabriel SE, Gakidou E, Gaspari F, Gillum RF, Gonzalez-Medina D, Halasa YA, Haring D, Harrison JE, Havmoeller R, Hay RJ, Hoen B, Hotez PJ, Hoy D, Jacobsen KH, James SL, Jasrasaria R, Jayaraman S, Johns N, Karthikeyan G, Kassebaum N, Keren A, Khoo JP, Knowlton LM, Kobusingye O, Koranteng A, Krishnamurthi R, Lipnick M, Lipshultz SE, Ohno SL, Mabweijano J, MacIntyre MF, Mallinger L, March L, Marks GB, Marks R, Matsumori A, Matzopoulos R, Mayosi BM, McAnulty JH, McDermott MM, McGrath J, Mensah GA, Merriman TR, Michaud C, Miller M, Miller TR, Mock C, Mocumbi AO, Mokdad AA, Moran A, Mulholland K, Nair MN, Naldi L, Narayan KM, Nasseri K, Norman P, O'Donnell M, Omer SB, Ortblad K, Osborne R, Ozgediz D, Pahari B, Pandian JD, Rivero AP, Padilla RP, Perez-Ruiz F, Perico N, Phillips D, Pierce K, Pope CA 3rd, Porrini E, Pourmalek F, Raju M, Ranganathan D, Rehm JT, Rein DB, Remuzzi G, Rivara FP, Roberts T, De León FR, Rosenfeld LC, Rushton L, Sacco RL, Salomon JA, Sampson U, Sanman E, Schwebel DC, Segui-Gomez M, Shepard DS, Singh D, Singleton J, Sliwa K, Smith E, Steer A, Taylor JA, Thomas B, Tleyjeh IM, Towbin JA, Truelsen T, Undurraga EA, Venketasubramanian N, Vijayakumar L, Vos T, Wagner GR, Wang M, Wang W, Watt K, Weinstock MA, Weintraub R, Wilkinson JD, Woolf AD, Wulf S, Yeh PH, Yip P, Zabetian A, Zheng ZJ, Lopez AD, Murray CJ, AlMazroa MA, Memish ZA, 2012. Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet 380: 2095–2128.
-
2.
Fotedar R, Stark D, Beebe N, Marriott D, Ellis J, Harkness J, 2007. Laboratory diagnostic techniques for Entamoeba species. Clin Microbiol Rev 20: 511–532.
-
3.
Goni P, Martin B, Villacampa M, Garcia A, Seral C, Castillo FJ, Clavel A, 2012. Evaluation of an immunochromatographic dip strip test for simultaneous detection of Cryptosporidium spp., Giardia duodenalis, and Entamoeba histolytica antigens in human faecal samples. Eur J Clin Microbiol Infect Dis 31: 2077–2082.
-
4.
Haque R, Ali IK, Akther S, Petri WA Jr, 1998. Comparison of PCR, isoenzyme analysis, and antigen detection for diagnosis of Entamoeba histolytica infection. J Clin Microbiol 36: 449–452.
-
5.
Pillai DR, Keystone JS, Sheppard DC, MacLean JD, MacPherson DW, Kain KC, 1999. Entamoeba histolytica and Entamoeba dispar: epidemiology and comparison of diagnostic methods in a setting of nonendemicity. Clin Infect Dis 29: 1315–1318.
-
6.
Mirelman D, Nuchamowitz Y, Stolarsky T, 1997. Comparison of use of enzyme-linked immunosorbent assay-based kits and PCR amplification of rRNA genes for simultaneous detection of Entamoeba histolytica and E. dispar. J Clin Microbiol 35: 2405–2407.
-
7.
Roy S, Kabir M, Mondal D, Ali IK, Petri WA Jr, Haque R, 2005. Real-time-PCR assay for diagnosis of Entamoeba histolytica infection. J Clin Microbiol 43: 2168–2172.
-
8.
Blessmann J, Buss H, Nu PA, Dinh BT, Ngo QT, Van AL, Alla MD, Jackson TF, Ravdin JI, Tannich E, 2002. Real-time PCR for detection and differentiation of Entamoeba histolytica and Entamoeba dispar in fecal samples. J Clin Microbiol 40: 4413–4417.
-
9.
Townes JM, Quick R, Gonzales OY, Linares M, Damiani E, Bopp CA, Wahlquist SP, Hutwagner LC, Hanover E, Mintz ED, Tauxe RV, 1997. Etiology of bloody diarrhea in Bolivian children: implications for empiric therapy. Bolivian Dysentery Study Group. J Infect Dis 175: 1527–1530.
-
10.
Bansal D, Sehgal R, Chawla Y, Mahajan RC, Malla N, 2004. In vitro activity of antiamoebic drugs against clinical isolates of Entamoeba histolytica and Entamoeba dispar. Ann Clin Microbiol Antimicrob 3: 27.
-
11.
Yan L, Zhou J, Zheng Y, Gamson AS, Roembke BT, Nakayama S, Sintim HO, 2014. Isothermal amplified detection of DNA and RNA. Mol Biosyst 10: 970–1003.
-
12.
Piepenburg O, Williams CH, Stemple DL, Armes NA, 2006. DNA detection using recombination proteins. PLoS Biol 4: e204.
-
13.
Crannell ZA, Castellanos-Gonzalez A, Irani A, Rohrman B, White AC, Richards-Kortum R, 2014. Nucleic acid test to diagnose cryptosporidiosis: lab assessment in animal and patient specimens. Anal Chem 86: 2565–2571.
-
14.
Crannell ZA, Cabada MM, Castellanos-Gonzalez A, Irani A, White AC, Richards-Kortum R, 2015. Recombinase polymerase amplification-based assay to diagnose Giardia in stool samples. Am J Trop Med Hyg 92: 583–587.
-
15.
Hamzah Z, Petmitr S, Mungthin M, Leelayoova S, Chavalitshewinkoon-Petmitr P, 2006. Differential detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii by a single-round PCR assay. J Clin Microbiol 44: 3196–3200.
-
16.
Gonin P, Trudel L, 2003. Detection and differentiation of Entamoeba histolytica and Entamoeba dispar isolates in clinical samples by PCR and enzyme-linked immunosorbent assay. J Clin Microbiol 41: 237–241.
-
17.
Hamzah Z, Petmitr S, Mungthin M, Leelayoova S, Chavalitshewinkoon-Petmitr P, 2010. Development of multiplex real-time polymerase chain reaction for detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in clinical specimens. Am J Trop Med Hyg 83: 909–913.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 77 | 77 | 19 |
| Full Text Views | 642 | 168 | 1 |
| PDF Downloads | 254 | 55 | 0 |
Detection of Entamoeba histolytica by Recombinase Polymerase Amplification
Gayatri Nair
Gayatri Nair
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
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Mauricio Rebolledo
Mauricio Rebolledo
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
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A. Clinton White Jr.
A. Clinton White Jr.
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
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Zachary Crannell
Zachary Crannell
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
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R. Rebecca Richards-Kortum
R. Rebecca Richards-Kortum
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
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A. Elizabeth Pinilla
A. Elizabeth Pinilla
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
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Juan David Ramírez
Juan David Ramírez
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
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M. Consuelo López
M. Consuelo López
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
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Alejandro Castellanos-Gonzalez
Alejandro Castellanos-Gonzalez
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
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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
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.
ProCite
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BibTeX
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-
1.
Lozano R, Naghavi M, Foreman K, Lim S, Shibuya K, Aboyans V, Abraham J, Adair T, Aggarwal R, Ahn SY, Alvarado M, Anderson HR, Anderson LM, Andrews KG, Atkinson C, Baddour LM, Barker-Collo S, Bartels DH, Bell ML, Benjamin EJ, Bennett D, Bhalla K, Bikbov B, Bin Abdulhak A, Birbeck G, Blyth F, Bolliger I, Boufous S, Bucello C, Burch M, Burney P, Carapetis J, Chen H, Chou D, Chugh SS, Coffeng LE, Colan SD, Colquhoun S, Colson KE, Condon J, Connor MD, Cooper LT, Corriere M, Cortinovis M, de Vaccaro KC, Couser W, Cowie BC, Criqui MH, Cross M, Dabhadkar KC, Dahodwala N, De Leo D, Degenhardt L, Delossantos A, Denenberg J, Des Jarlais DC, Dharmaratne SD, Dorsey ER, Driscoll T, Duber H, Ebel B, Erwin PJ, Espindola P, Ezzati M, Feigin V, Flaxman AD, Forouzanfar MH, Fowkes FG, Franklin R, Fransen M, Freeman MK, Gabriel SE, Gakidou E, Gaspari F, Gillum RF, Gonzalez-Medina D, Halasa YA, Haring D, Harrison JE, Havmoeller R, Hay RJ, Hoen B, Hotez PJ, Hoy D, Jacobsen KH, James SL, Jasrasaria R, Jayaraman S, Johns N, Karthikeyan G, Kassebaum N, Keren A, Khoo JP, Knowlton LM, Kobusingye O, Koranteng A, Krishnamurthi R, Lipnick M, Lipshultz SE, Ohno SL, Mabweijano J, MacIntyre MF, Mallinger L, March L, Marks GB, Marks R, Matsumori A, Matzopoulos R, Mayosi BM, McAnulty JH, McDermott MM, McGrath J, Mensah GA, Merriman TR, Michaud C, Miller M, Miller TR, Mock C, Mocumbi AO, Mokdad AA, Moran A, Mulholland K, Nair MN, Naldi L, Narayan KM, Nasseri K, Norman P, O'Donnell M, Omer SB, Ortblad K, Osborne R, Ozgediz D, Pahari B, Pandian JD, Rivero AP, Padilla RP, Perez-Ruiz F, Perico N, Phillips D, Pierce K, Pope CA 3rd, Porrini E, Pourmalek F, Raju M, Ranganathan D, Rehm JT, Rein DB, Remuzzi G, Rivara FP, Roberts T, De León FR, Rosenfeld LC, Rushton L, Sacco RL, Salomon JA, Sampson U, Sanman E, Schwebel DC, Segui-Gomez M, Shepard DS, Singh D, Singleton J, Sliwa K, Smith E, Steer A, Taylor JA, Thomas B, Tleyjeh IM, Towbin JA, Truelsen T, Undurraga EA, Venketasubramanian N, Vijayakumar L, Vos T, Wagner GR, Wang M, Wang W, Watt K, Weinstock MA, Weintraub R, Wilkinson JD, Woolf AD, Wulf S, Yeh PH, Yip P, Zabetian A, Zheng ZJ, Lopez AD, Murray CJ, AlMazroa MA, Memish ZA, 2012. Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet 380: 2095–2128.
-
2.
Fotedar R, Stark D, Beebe N, Marriott D, Ellis J, Harkness J, 2007. Laboratory diagnostic techniques for Entamoeba species. Clin Microbiol Rev 20: 511–532.
-
3.
Goni P, Martin B, Villacampa M, Garcia A, Seral C, Castillo FJ, Clavel A, 2012. Evaluation of an immunochromatographic dip strip test for simultaneous detection of Cryptosporidium spp., Giardia duodenalis, and Entamoeba histolytica antigens in human faecal samples. Eur J Clin Microbiol Infect Dis 31: 2077–2082.
-
4.
Haque R, Ali IK, Akther S, Petri WA Jr, 1998. Comparison of PCR, isoenzyme analysis, and antigen detection for diagnosis of Entamoeba histolytica infection. J Clin Microbiol 36: 449–452.
-
5.
Pillai DR, Keystone JS, Sheppard DC, MacLean JD, MacPherson DW, Kain KC, 1999. Entamoeba histolytica and Entamoeba dispar: epidemiology and comparison of diagnostic methods in a setting of nonendemicity. Clin Infect Dis 29: 1315–1318.
-
6.
Mirelman D, Nuchamowitz Y, Stolarsky T, 1997. Comparison of use of enzyme-linked immunosorbent assay-based kits and PCR amplification of rRNA genes for simultaneous detection of Entamoeba histolytica and E. dispar. J Clin Microbiol 35: 2405–2407.
-
7.
Roy S, Kabir M, Mondal D, Ali IK, Petri WA Jr, Haque R, 2005. Real-time-PCR assay for diagnosis of Entamoeba histolytica infection. J Clin Microbiol 43: 2168–2172.
-
8.
Blessmann J, Buss H, Nu PA, Dinh BT, Ngo QT, Van AL, Alla MD, Jackson TF, Ravdin JI, Tannich E, 2002. Real-time PCR for detection and differentiation of Entamoeba histolytica and Entamoeba dispar in fecal samples. J Clin Microbiol 40: 4413–4417.
-
9.
Townes JM, Quick R, Gonzales OY, Linares M, Damiani E, Bopp CA, Wahlquist SP, Hutwagner LC, Hanover E, Mintz ED, Tauxe RV, 1997. Etiology of bloody diarrhea in Bolivian children: implications for empiric therapy. Bolivian Dysentery Study Group. J Infect Dis 175: 1527–1530.
-
10.
Bansal D, Sehgal R, Chawla Y, Mahajan RC, Malla N, 2004. In vitro activity of antiamoebic drugs against clinical isolates of Entamoeba histolytica and Entamoeba dispar. Ann Clin Microbiol Antimicrob 3: 27.
-
11.
Yan L, Zhou J, Zheng Y, Gamson AS, Roembke BT, Nakayama S, Sintim HO, 2014. Isothermal amplified detection of DNA and RNA. Mol Biosyst 10: 970–1003.
-
12.
Piepenburg O, Williams CH, Stemple DL, Armes NA, 2006. DNA detection using recombination proteins. PLoS Biol 4: e204.
-
13.
Crannell ZA, Castellanos-Gonzalez A, Irani A, Rohrman B, White AC, Richards-Kortum R, 2014. Nucleic acid test to diagnose cryptosporidiosis: lab assessment in animal and patient specimens. Anal Chem 86: 2565–2571.
-
14.
Crannell ZA, Cabada MM, Castellanos-Gonzalez A, Irani A, White AC, Richards-Kortum R, 2015. Recombinase polymerase amplification-based assay to diagnose Giardia in stool samples. Am J Trop Med Hyg 92: 583–587.
-
15.
Hamzah Z, Petmitr S, Mungthin M, Leelayoova S, Chavalitshewinkoon-Petmitr P, 2006. Differential detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii by a single-round PCR assay. J Clin Microbiol 44: 3196–3200.
-
16.
Gonin P, Trudel L, 2003. Detection and differentiation of Entamoeba histolytica and Entamoeba dispar isolates in clinical samples by PCR and enzyme-linked immunosorbent assay. J Clin Microbiol 41: 237–241.
-
17.
Hamzah Z, Petmitr S, Mungthin M, Leelayoova S, Chavalitshewinkoon-Petmitr P, 2010. Development of multiplex real-time polymerase chain reaction for detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in clinical specimens. Am J Trop Med Hyg 83: 909–913.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 77 | 77 | 19 |
| Full Text Views | 642 | 168 | 1 |
| PDF Downloads | 254 | 55 | 0 |