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Reference Manager
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-
1.
Haque R, Mondal D, Karim A, Molla IH, Rahim A, Faruque AS, Ahmad N, Kirkpatrick BD, Houpt E, Snider C, Petri WA Jr, 2009. Prospective case-control study of the association between common enteric protozoal parasites and diarrhea in Bangladesh. Clin Infect Dis 48: 1191–1197.
-
2.
ten Hove RJ, van Esbroeck M, Vervoort T, van den Ende J, van Lieshout L, Verweij JJ, 2009. Molecular diagnostics of intestinal parasites in returning travellers. Eur J Clin Microbiol Infect Dis 28: 1045–1053.
-
3.
Basuni M, Muhi J, Othman N, Verweij JJ, Ahmad M, Miswan N, Rahumatullah A, Aziz FA, Zainudin NS, Noordin R, 2011. A pentaplex real-time polymerase chain reaction assay for detection of four species of soil-transmitted helminths. Am J Trop Med Hyg 84: 338–343.
-
4.
Pecson BM, Barrios JA, Johnson DR, Nelson KL, 2006. A real-time PCR method for quantifying viable Ascaris eggs using the first internally transcribed spacer region of ribosomal DNA. Appl Environ Microbiol 72: 7864–7872.
-
5.
Wolk DM, Schneider SK, Wengenack NL, Sloan LM, Rosenblatt JE, 2002. Real-time PCR method for detection of Encephalitozoon intestinalis from stool specimens. J Clin Microbiol 40: 3922–3928.
-
6.
Krolewiecki AJ, Ramanathan R, Fink V, McAuliffe I, Cajal SP, Won K, Juarez M, Di Paolo A, Tapia L, Acosta N, Lee R, Lammie P, Abraham D, Nutman TB, 2010. Improved diagnosis of Strongyloides stercoralis using recombinant antigen-based serologies in a community-wide study in northern Argentina. Clin Vaccine Immunol 17: 1624–1630.
-
7.
Haque R, Neville LM, Hahn P, Petri WA Jr, 1995. Rapid diagnosis of Entamoeba infection by using Entamoeba and Entamoeba histolytica stool antigen detection kits. J Clin Microbiol 33: 2558–2561.
-
8.
Petri WA Jr, 2000. Protozoan parasites that infect the gastrointestinal tract. Cur Opin Gastroenterol 16: 18–23.
-
9.
Haque R, Kabir M, Noor Z, Rahman SM, Mondal D, Alam F, Rahman I, Al Mahmood A, Ahmed N, Petri WA Jr, 2010. Diagnosis of amebic liver abscess and amebic colitis by detection of Entamoeba histolytica DNA in blood, urine, and saliva by a real-time PCR assay. J Clin Microbiol 48: 2798–2801.
-
10.
Cooper PJ, Chico ME, Guadalupe I, Sandoval CA, Mitre E, Platts-Mills TA, Barreto ML, Rodrigues LC, Strachan DP, Griffin GE, 2011. Impact of early life exposures to geohelminth infections on the development of vaccine immunity, allergic sensitization, and allergic inflammatory diseases in children living in tropical Ecuador: the ECUAVIDA birth cohort study. BMC Infect Dis 11: 184.
-
11.
Larson D, Cooper PJ, Hubner MP, Reyes J, Vaca M, Chico M, Kong HH, Mitre E, 2012. Helminth infection is associated with decreased basophil responsiveness in human beings. J Allergy Clin Immunol 130: 270–272.
-
12.
da Silva AJ, Bornay-Llinares FJ, Moura IN, Slemenda SB, Tutttle JL, Pieniazek NJ, 1999. Fast and reliable extraction of protozoan parasite DNA from fecal specimens. Mol Diagn 4: 57–64.
-
13.
Carlson JR, Sullivan PS, Harry DJ, Stork MA, Thorton SA, DuPont HL, 1988. Enzyme immunoassay for the detection of Giardia lamblia. Eur J Clin Microbiol Infect Dis 7: 538–540.
-
14.
Hotez P, 2011. Enlarging the “audacious goal”: elimination of the world's high prevalence neglected tropical diseases. Vaccine 29 (Suppl 4): D104–D110.
-
15.
Tejman-Yarden N, Eckmann L, 2011. New approaches to the treatment of giardiasis. Curr Opin Infect Dis 24: 451–456.
-
16.
Abu Al-Soud W, Radstrom P, 1998. Capacity of nine thermostable DNA polymerases to mediate DNA amplification in the presence of PCR-inhibiting samples. Appl Environ Microbiol 64: 3748–3753.
-
17.
Koonjul PK, Brandt WF, Farrant JM, Lindsey GG, 1999. Inclusion of polyvinylpyrrolidone in the polymerase chain reaction reverses the inhibitory effects of polyphenolic contamination of RNA. Nucleic Acids Res 27: 915–916.
-
18.
Fahle GA, Fischer SH, 2000. Comparison of six commercial DNA extraction kits for recovery of cytomegalovirus DNA from spiked human specimens. J Clin Microbiol 38: 3860–3863.
-
19.
Hotez PJ, Brooker S, Bethony JM, Bottazzi ME, Loukas A, Xiao S, 2004. Hookworm infection. N Engl J Med 351: 799–807.
-
20.
Bercu TE, Petri WA, Behm JW, 2007. Amebic colitis: new insights into pathogenesis and treatment. Curr Gastroenterol Rep 9: 429–433.
-
21.
Ungar BL, Soave R, Fayer R, Nash TE, 1986. Enzyme immunoassay detection of immunoglobulin M and G antibodies to Cryptosporidium in immunocompetent and immunocompromised persons. J Infect Dis 153: 570–578.
-
22.
Hoxie NJ, Davis JP, Vergeront JM, Nashold RD, Blair KA, 1997. Cryptosporidiosis-associated mortality following a massive waterborne outbreak in Milwaukee, Wisconsin. Am J Public Health 87: 2032–2035.
-
23.
Nash TE, Herrington DA, Losonsky GA, Levine MM, 1987. Experimental human infections with Giardia lamblia. J Infect Dis 156: 974–984.
-
24.
Hall A, Nahar Q, 1993. Albendazole as a treatment for infections with Giardia duodenalis in children in Bangladesh. Trans R Soc Trop Med Hyg 87: 84–86.
-
25.
Basuni M, Mohamed Z, Ahmad M, Zakaria NZ, Noordin R, 2012. Detection of selected intestinal helminths and protozoa at Hospital Universiti Sains Malaysia using multiplex real-time PCR. Trop Biomed 29: 434–442.
-
26.
Petri WA Jr, Miller M, Binder HJ, Levine MM, Dillingham R, Guerrant RL, 2008. Enteric infections, diarrhea, and their impact on function and development. J Clin Invest 118: 1277–1290.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 3851 | 2750 | 94 |
| Full Text Views | 1847 | 33 | 5 |
| PDF Downloads | 1004 | 43 | 6 |
A Novel, Multi-Parallel, Real-Time Polymerase Chain Reaction Approach for Eight Gastrointestinal Parasites Provides Improved Diagnostic Capabilities to Resource-Limited At-Risk Populations
Rojelio Mejia
Rojelio Mejia
Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland; Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador; Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Quito, Ecuador; Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Yosselin Vicuña
Yosselin Vicuña
Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland; Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador; Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Quito, Ecuador; Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Nely Broncano
Nely Broncano
Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland; Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador; Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Quito, Ecuador; Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Carlos Sandoval
Carlos Sandoval
Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland; Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador; Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Quito, Ecuador; Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Maritza Vaca
Maritza Vaca
Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland; Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador; Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Quito, Ecuador; Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Martha Chico
Martha Chico
Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland; Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador; Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Quito, Ecuador; Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Philip J. Cooper
Philip J. Cooper
Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland; Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador; Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Quito, Ecuador; Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Thomas B. Nutman
Thomas B. Nutman
Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland; Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador; Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Quito, Ecuador; Molecular and Biochemical Parasitology, Liverpool School of Tropical Medicine, Liverpool, United Kingdom
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Diagnosis of gastrointestinal parasites has traditionally relied on stool microscopy, which has low diagnostic sensitivity and specificity. We have developed a novel, rapid, high-throughput quantitative multi-parallel real-time polymerase chain reaction (qPCR) platform. Species-specific primers/probes were used for eight common gastrointestinal parasite pathogens: Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale, Giardia lamblia, Cryptosporidium spp., Entamoeba histolytica, Trichuris trichiura, and Strongyloides stercoralis. Stool samples from 400 13-month-old children in rural Ecuador were analyzed and the qPCR was compared with a standard direct wet mount slide for stool microscopy, as were 125 8–14-year-old children before and after anthelmintic treatment. The qPCR showed higher detection rates for all parasites compared with direct microscopy, Ascaris (7.0% versus 5.5%) and for Giardia (31.5% versus 5.8%). Using an enhanced DNA extraction method, we were able to detect T. trichiura DNA. These assays will be useful to refine treatment options for affected populations, ultimately leading to better health outcomes.
Author Notes
Authors' addresses: Rojelio Mejia, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, E-mail: rojelio.mejia@bcm.edu. Thomas B. Nutman, Helminth Immunology Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, E-mail: tnutman@niaid.nih.gov. Yosselin Vicuña, Nely Broncano, Carlos Sandoval, Maritza Vaca, Martha Chico, and Philip J. Cooper, Colegio de Ciencias de la Salud, Universidad San Francisco de Quito, Fundación Ecuatoriana para la Investigación en Salud, Quito, Ecuador, E-mails: yossvi@gmail.com, nelyes@hotmail.es, sandoval_acarlos@hotmail.com, marimar_ecq@yahoo.com, marthachico6@yahoo.es, and p.j.cooper@liverpool.ac.uk.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Haque R, Mondal D, Karim A, Molla IH, Rahim A, Faruque AS, Ahmad N, Kirkpatrick BD, Houpt E, Snider C, Petri WA Jr, 2009. Prospective case-control study of the association between common enteric protozoal parasites and diarrhea in Bangladesh. Clin Infect Dis 48: 1191–1197.
-
2.
ten Hove RJ, van Esbroeck M, Vervoort T, van den Ende J, van Lieshout L, Verweij JJ, 2009. Molecular diagnostics of intestinal parasites in returning travellers. Eur J Clin Microbiol Infect Dis 28: 1045–1053.
-
3.
Basuni M, Muhi J, Othman N, Verweij JJ, Ahmad M, Miswan N, Rahumatullah A, Aziz FA, Zainudin NS, Noordin R, 2011. A pentaplex real-time polymerase chain reaction assay for detection of four species of soil-transmitted helminths. Am J Trop Med Hyg 84: 338–343.
-
4.
Pecson BM, Barrios JA, Johnson DR, Nelson KL, 2006. A real-time PCR method for quantifying viable Ascaris eggs using the first internally transcribed spacer region of ribosomal DNA. Appl Environ Microbiol 72: 7864–7872.
-
5.
Wolk DM, Schneider SK, Wengenack NL, Sloan LM, Rosenblatt JE, 2002. Real-time PCR method for detection of Encephalitozoon intestinalis from stool specimens. J Clin Microbiol 40: 3922–3928.
-
6.
Krolewiecki AJ, Ramanathan R, Fink V, McAuliffe I, Cajal SP, Won K, Juarez M, Di Paolo A, Tapia L, Acosta N, Lee R, Lammie P, Abraham D, Nutman TB, 2010. Improved diagnosis of Strongyloides stercoralis using recombinant antigen-based serologies in a community-wide study in northern Argentina. Clin Vaccine Immunol 17: 1624–1630.
-
7.
Haque R, Neville LM, Hahn P, Petri WA Jr, 1995. Rapid diagnosis of Entamoeba infection by using Entamoeba and Entamoeba histolytica stool antigen detection kits. J Clin Microbiol 33: 2558–2561.
-
8.
Petri WA Jr, 2000. Protozoan parasites that infect the gastrointestinal tract. Cur Opin Gastroenterol 16: 18–23.
-
9.
Haque R, Kabir M, Noor Z, Rahman SM, Mondal D, Alam F, Rahman I, Al Mahmood A, Ahmed N, Petri WA Jr, 2010. Diagnosis of amebic liver abscess and amebic colitis by detection of Entamoeba histolytica DNA in blood, urine, and saliva by a real-time PCR assay. J Clin Microbiol 48: 2798–2801.
-
10.
Cooper PJ, Chico ME, Guadalupe I, Sandoval CA, Mitre E, Platts-Mills TA, Barreto ML, Rodrigues LC, Strachan DP, Griffin GE, 2011. Impact of early life exposures to geohelminth infections on the development of vaccine immunity, allergic sensitization, and allergic inflammatory diseases in children living in tropical Ecuador: the ECUAVIDA birth cohort study. BMC Infect Dis 11: 184.
-
11.
Larson D, Cooper PJ, Hubner MP, Reyes J, Vaca M, Chico M, Kong HH, Mitre E, 2012. Helminth infection is associated with decreased basophil responsiveness in human beings. J Allergy Clin Immunol 130: 270–272.
-
12.
da Silva AJ, Bornay-Llinares FJ, Moura IN, Slemenda SB, Tutttle JL, Pieniazek NJ, 1999. Fast and reliable extraction of protozoan parasite DNA from fecal specimens. Mol Diagn 4: 57–64.
-
13.
Carlson JR, Sullivan PS, Harry DJ, Stork MA, Thorton SA, DuPont HL, 1988. Enzyme immunoassay for the detection of Giardia lamblia. Eur J Clin Microbiol Infect Dis 7: 538–540.
-
14.
Hotez P, 2011. Enlarging the “audacious goal”: elimination of the world's high prevalence neglected tropical diseases. Vaccine 29 (Suppl 4): D104–D110.
-
15.
Tejman-Yarden N, Eckmann L, 2011. New approaches to the treatment of giardiasis. Curr Opin Infect Dis 24: 451–456.
-
16.
Abu Al-Soud W, Radstrom P, 1998. Capacity of nine thermostable DNA polymerases to mediate DNA amplification in the presence of PCR-inhibiting samples. Appl Environ Microbiol 64: 3748–3753.
-
17.
Koonjul PK, Brandt WF, Farrant JM, Lindsey GG, 1999. Inclusion of polyvinylpyrrolidone in the polymerase chain reaction reverses the inhibitory effects of polyphenolic contamination of RNA. Nucleic Acids Res 27: 915–916.
-
18.
Fahle GA, Fischer SH, 2000. Comparison of six commercial DNA extraction kits for recovery of cytomegalovirus DNA from spiked human specimens. J Clin Microbiol 38: 3860–3863.
-
19.
Hotez PJ, Brooker S, Bethony JM, Bottazzi ME, Loukas A, Xiao S, 2004. Hookworm infection. N Engl J Med 351: 799–807.
-
20.
Bercu TE, Petri WA, Behm JW, 2007. Amebic colitis: new insights into pathogenesis and treatment. Curr Gastroenterol Rep 9: 429–433.
-
21.
Ungar BL, Soave R, Fayer R, Nash TE, 1986. Enzyme immunoassay detection of immunoglobulin M and G antibodies to Cryptosporidium in immunocompetent and immunocompromised persons. J Infect Dis 153: 570–578.
-
22.
Hoxie NJ, Davis JP, Vergeront JM, Nashold RD, Blair KA, 1997. Cryptosporidiosis-associated mortality following a massive waterborne outbreak in Milwaukee, Wisconsin. Am J Public Health 87: 2032–2035.
-
23.
Nash TE, Herrington DA, Losonsky GA, Levine MM, 1987. Experimental human infections with Giardia lamblia. J Infect Dis 156: 974–984.
-
24.
Hall A, Nahar Q, 1993. Albendazole as a treatment for infections with Giardia duodenalis in children in Bangladesh. Trans R Soc Trop Med Hyg 87: 84–86.
-
25.
Basuni M, Mohamed Z, Ahmad M, Zakaria NZ, Noordin R, 2012. Detection of selected intestinal helminths and protozoa at Hospital Universiti Sains Malaysia using multiplex real-time PCR. Trop Biomed 29: 434–442.
-
26.
Petri WA Jr, Miller M, Binder HJ, Levine MM, Dillingham R, Guerrant RL, 2008. Enteric infections, diarrhea, and their impact on function and development. J Clin Invest 118: 1277–1290.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 3851 | 2750 | 94 |
| Full Text Views | 1847 | 33 | 5 |
| PDF Downloads | 1004 | 43 | 6 |