ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
GBD 2016 Causes of Death Collaborators, 2017. Global, regional, and national age-sex specific mortality for 264 causes of death, 1980–2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet 390: 1151–1210.
-
2.
Kotloff KL et al. 2013. Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet 382: 209–222.
-
3.
Platts-Mills JA et al. 2015. Pathogen-specific burdens of community diarrhoea in developing countries: a multisite birth cohort study (MAL-ED). Lancet Glob Health 3: e564–e575.
-
4.
Nakano T et al. 1990. Survey of enteropathogenic agents in children with and without diarrhoea in Ghana. J Trop Med Hyg 93: 408–412.
-
5.
Nimri LF, Meqdam M, 2004. Enteropathogens associated with cases of gastroenteritis in a rural population in Jordan. Clin Microbiol Infect 10: 634–639.
-
6.
Rogawski ET et al. MAL-ED Network Investigators, 2017. Determinants and impact of Giardia infection in the first 2 years of life in the MAL-ED birth cohort. J Pediatr Infect Dis Soc 6: 153–160.
-
7.
Fischer Walker CL, Sack D, Black RE, 2010. Etiology of diarrhea in older children, adolescents and adults: a systematic review. PLoS Negl Trop Dis 4: e768.
-
8.
Humphries RM, Linscott AJ, 2015. Laboratory diagnosis of bacterial gastroenteritis. Clin Microbiol Rev 28: 3–31.
-
9.
Alam M et al. 2010. Diagnostic limitations to accurate diagnosis of cholera. J Clin Microbiol 48: 3918–3922.
-
10.
Guerrant RL, Shields DS, Thorson SM, Schorling JB, Groschel DH, 1985. Evaluation and diagnosis of acute infectious diarrhea. Am J Med 78: 91–98.
-
11.
Platts-Mills JA, Liu J, Houpt ER, 2013. New concepts in diagnostics for infectious diarrhea. Mucosal Immunol 6: 876–885.
-
12.
Platts-Mills JA, Operario DJ, Houpt ER, 2012. Molecular diagnosis of diarrhea: current status and future potential. Curr Infect Dis Rep 14: 41–46.
-
13.
Pawlowski SW, Warren CA, Guerrant R, 2009. Diagnosis and treatment of acute or persistent diarrhea. Gastroenterology 136: 1874–1886.
-
14.
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.
-
15.
Ley B et al. 2012. Evaluation of a rapid dipstick (Crystal VC) for the diagnosis of cholera in Zanzibar and a comparison with previous studies. PLoS one 7: e36930.
-
16.
Sinha A et al. 2012. Evaluation of a rapid dipstick test for identifying cholera cases during the outbreak. Indian J Med Res 135: 523–528.
-
17.
George CM et al. 2014. Evaluation of enrichment method for the detection of Vibrio cholerae O1 using a rapid dipstick test in Bangladesh. Trop Med Int Health 19: 301–307.
-
18.
Haque R, Roy S, Siddique A, Mondal U, Rahman SM, Mondal D, Houpt E, Petri WA Jr., 2007. Multiplex real-time PCR assay for detection of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. Am J Trop Med Hyg 76: 713–717.
-
19.
Liu J et al. 2014. Development and assessment of molecular diagnostic tests for 15 enteropathogens causing childhood diarrhoea: a multicentre study. Lancet Infect Dis 14: 716–724.
-
20.
Liu J et al. 2016. Optimization of quantitative PCR methods for enteropathogen detection. PLoS One 11: e0158199.
-
21.
Debes AK et al. 2016. Evaluation in Cameroon of a novel, simplified methodology to assist molecular microbiological analysis of V. cholerae in resource-limited settings. PLoS Negl Trop Dis 10: e0004307.
-
22.
Lalani T, Tisdale MD, Maguire JD, Wongsrichanalai C, Riddle MS, Tribble DR, 2015. Detection of enteropathogens associated with travelers’ diarrhea using a multiplex Luminex-based assay performed on stool samples smeared on Whatman FTA Elute cards. Diagn Microbiol Infect Dis 83: 18–20.
-
23.
Liu J et al. 2016. Use of quantitative molecular diagnostic methods to identify causes of diarrhoea in children: a reanalysis of the GEMS case-control study. Lancet 388: 1291–1301.
-
24.
Debes AK, Ateudjieu J, Guenou E, Ebile W, Sonkoua IT, Njimbia AC, Steinwald P, Ram M, Sack DA, 2016. Clinical and environmental surveillance for Vibrio cholerae in resource constrained areas: application during a 1-year surveillance in the far north region of Cameroon. Am J Trop Med Hyg 94: 537–543.
-
25.
Lothigius A, Janzon A, Begum Y, Sjöling A, Qadri F, Svennerholm AM, Bölin I, 2008. Enterotoxigenic Escherichia coli is detectable in water samples from an endemic area by real-time PCR. J Appl Microbiol 104: 1128–1136.
-
26.
Katevatis C, Fan A, Klapperich CM, 2017. Low concentration DNA extraction and recovery using a silica solid phase. PLoS One 12: e0176848.
-
27.
Angelakis E, Bachar D, Henrissat B, Armougom F, Audoly G, Lagier JC, Robert C, Raoult D, 2016. Glycans affect DNA extraction and induce substantial differences in gut metagenomic studies. Sci Rep 6: 26276.
-
28.
Holland JL, Louie L, Simor AE, Louie M, 2000. PCR detection of Escherichia coli O157:H7 directly from stools: evaluation of commercial extraction methods for purifying fecal DNA. J Clin Microbiol 38: 4108–4113.
-
29.
Oikarinen S, Tauriainen S, Viskari H, Simell O, Knip M, Virtanen S, Hyöty H, 2009. PCR inhibition in stool samples in relation to age of infants. J Clin Virol 44: 211–214.
-
30.
Widjojoatmodjo MN, Fluit AC, Torensma R, Verdonk GP, Verhoef J, 1992. The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples. J Clin Microbiol 30: 3195–3199.
-
31.
Liu J et al. 2013. A laboratory-developed TaqMan array card for simultaneous detection of 19 enteropathogens. J Clin Microbiol 51: 472–480.
-
32.
Nichols RA, Smith HV, 2004. Optimization of DNA extraction and molecular detection of Cryptosporidium oocysts in natural mineral water sources. J Food Prot 67: 524–532.
-
33.
Robertson LJ, Campbell AT, Smith HV, 1992. Survival of Cryptosporidium parvum oocysts under various environmental pressures. Appl Environ Microbiol 58: 3494–3500.
-
34.
Kaucner C, Stinear T, 1998. Sensitive and rapid detection of viable Giardia cysts and Cryptosporidium parvum oocysts in large-volume water samples with wound fiberglass cartridge filters and reverse transcription-PCR. Appl Environ Microbiol 64: 1743–1749.
-
35.
Mero S, Kirveskari J, Antikainen J, Ursing J, Rombo L, Kofoed PE, Kantele A, 2017. Multiplex PCR detection of Cryptosporidium sp, Giardia lamblia and Entamoeba histolytica directly from dried stool samples from Guinea-Bissauan children with diarrhoea. Infect Dis 49: 655–663.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 782 | 685 | 66 |
| Full Text Views | 579 | 12 | 2 |
| PDF Downloads | 238 | 12 | 2 |
Whatman Protein Saver Cards for Storage and Detection of Parasitic Enteropathogens
Gayathri Natarajan
Gayathri Natarajan
Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland;
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Mamun Kabir
Mamun Kabir
International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh
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Jamie Perin
Jamie Perin
Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland;
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Biplob Hossain
Biplob Hossain
International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh
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Amanda Debes
Amanda Debes
Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland;
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Rashidul Haque
Rashidul Haque
International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh
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Christine Marie George
Christine Marie George
Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland;
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Current methods to identify the etiology of diarrhea require laboratory facilities for storage of pathogens, which is often challenging in low-resource settings. This study evaluated the efficacy of a low-cost method for preserving stool specimens for the detection of parasitic enteropathogens using Whatman 903 protein saver cards (Sigma-Aldrich, St. Louis, MO). Stool samples known to be positive by multiplex real-time polymerase chain reaction for Giardia lamblia, Cryptosporidium spp., and Entamoeba histolytica parasites were preserved on 232 Whatman cards. DNA was then extracted from cards using Chelex and Qiagen extraction protocols, and tested for these parasites using multiplex real-time PCR. We included stool samples known to have a higher parasite load (cycle threshold [ct]-value < 30) and those with a lower parasite load (ct values 30–35). Sensitivities and specificities were determined using DNA extracted directly from whole stool samples using Qiagen kits (QIAGEN, Hilden, Germany). For whole stool samples with ct values < 30, preserved directly on Whatman 903 protein saver cards for Giardia analysis, the sensitivity was 100% for both Qiagen and Chelex DNA extraction. For E. histolytica, this was 100% for sensitivity for Qiagen and 80% for Chelex DNA extractions, and for Cryptosporidium, this was 80% for Qiagen and 50% for Chelex DNA extraction. The specificity was 100% for all parasites for all extraction procedures. Given the high sensitivity for stool samples with higher parasite loads, we recommend the use of the Whatman 903 protein saver card for preserving fecal specimens for the analysis of Giardia and E. histolytica using Qiagen DNA extractions in low-resource settings.
- Supplemental Materials (PDF 43 KB)
- Supplemental Materials (PDF 31 KB)
Author Notes
Authors’ addresses: Gayathri Natarajan, Jamie Perin, Amanda Debes, and Christine Marie George, Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, E-mails: gnatara1@jhmi.edu, jperin@jhu.edu, adebes1@jhu.edu, and cgeorg19@jhu.edu. Mamun Kabir, Biplob Hossain, and Rashidul Haque, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh, E-mails: mamunk@icddrb.org, bhossain@icddrb.org, and rhaque@icddrb.org.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
GBD 2016 Causes of Death Collaborators, 2017. Global, regional, and national age-sex specific mortality for 264 causes of death, 1980–2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet 390: 1151–1210.
-
2.
Kotloff KL et al. 2013. Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet 382: 209–222.
-
3.
Platts-Mills JA et al. 2015. Pathogen-specific burdens of community diarrhoea in developing countries: a multisite birth cohort study (MAL-ED). Lancet Glob Health 3: e564–e575.
-
4.
Nakano T et al. 1990. Survey of enteropathogenic agents in children with and without diarrhoea in Ghana. J Trop Med Hyg 93: 408–412.
-
5.
Nimri LF, Meqdam M, 2004. Enteropathogens associated with cases of gastroenteritis in a rural population in Jordan. Clin Microbiol Infect 10: 634–639.
-
6.
Rogawski ET et al. MAL-ED Network Investigators, 2017. Determinants and impact of Giardia infection in the first 2 years of life in the MAL-ED birth cohort. J Pediatr Infect Dis Soc 6: 153–160.
-
7.
Fischer Walker CL, Sack D, Black RE, 2010. Etiology of diarrhea in older children, adolescents and adults: a systematic review. PLoS Negl Trop Dis 4: e768.
-
8.
Humphries RM, Linscott AJ, 2015. Laboratory diagnosis of bacterial gastroenteritis. Clin Microbiol Rev 28: 3–31.
-
9.
Alam M et al. 2010. Diagnostic limitations to accurate diagnosis of cholera. J Clin Microbiol 48: 3918–3922.
-
10.
Guerrant RL, Shields DS, Thorson SM, Schorling JB, Groschel DH, 1985. Evaluation and diagnosis of acute infectious diarrhea. Am J Med 78: 91–98.
-
11.
Platts-Mills JA, Liu J, Houpt ER, 2013. New concepts in diagnostics for infectious diarrhea. Mucosal Immunol 6: 876–885.
-
12.
Platts-Mills JA, Operario DJ, Houpt ER, 2012. Molecular diagnosis of diarrhea: current status and future potential. Curr Infect Dis Rep 14: 41–46.
-
13.
Pawlowski SW, Warren CA, Guerrant R, 2009. Diagnosis and treatment of acute or persistent diarrhea. Gastroenterology 136: 1874–1886.
-
14.
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.
-
15.
Ley B et al. 2012. Evaluation of a rapid dipstick (Crystal VC) for the diagnosis of cholera in Zanzibar and a comparison with previous studies. PLoS one 7: e36930.
-
16.
Sinha A et al. 2012. Evaluation of a rapid dipstick test for identifying cholera cases during the outbreak. Indian J Med Res 135: 523–528.
-
17.
George CM et al. 2014. Evaluation of enrichment method for the detection of Vibrio cholerae O1 using a rapid dipstick test in Bangladesh. Trop Med Int Health 19: 301–307.
-
18.
Haque R, Roy S, Siddique A, Mondal U, Rahman SM, Mondal D, Houpt E, Petri WA Jr., 2007. Multiplex real-time PCR assay for detection of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. Am J Trop Med Hyg 76: 713–717.
-
19.
Liu J et al. 2014. Development and assessment of molecular diagnostic tests for 15 enteropathogens causing childhood diarrhoea: a multicentre study. Lancet Infect Dis 14: 716–724.
-
20.
Liu J et al. 2016. Optimization of quantitative PCR methods for enteropathogen detection. PLoS One 11: e0158199.
-
21.
Debes AK et al. 2016. Evaluation in Cameroon of a novel, simplified methodology to assist molecular microbiological analysis of V. cholerae in resource-limited settings. PLoS Negl Trop Dis 10: e0004307.
-
22.
Lalani T, Tisdale MD, Maguire JD, Wongsrichanalai C, Riddle MS, Tribble DR, 2015. Detection of enteropathogens associated with travelers’ diarrhea using a multiplex Luminex-based assay performed on stool samples smeared on Whatman FTA Elute cards. Diagn Microbiol Infect Dis 83: 18–20.
-
23.
Liu J et al. 2016. Use of quantitative molecular diagnostic methods to identify causes of diarrhoea in children: a reanalysis of the GEMS case-control study. Lancet 388: 1291–1301.
-
24.
Debes AK, Ateudjieu J, Guenou E, Ebile W, Sonkoua IT, Njimbia AC, Steinwald P, Ram M, Sack DA, 2016. Clinical and environmental surveillance for Vibrio cholerae in resource constrained areas: application during a 1-year surveillance in the far north region of Cameroon. Am J Trop Med Hyg 94: 537–543.
-
25.
Lothigius A, Janzon A, Begum Y, Sjöling A, Qadri F, Svennerholm AM, Bölin I, 2008. Enterotoxigenic Escherichia coli is detectable in water samples from an endemic area by real-time PCR. J Appl Microbiol 104: 1128–1136.
-
26.
Katevatis C, Fan A, Klapperich CM, 2017. Low concentration DNA extraction and recovery using a silica solid phase. PLoS One 12: e0176848.
-
27.
Angelakis E, Bachar D, Henrissat B, Armougom F, Audoly G, Lagier JC, Robert C, Raoult D, 2016. Glycans affect DNA extraction and induce substantial differences in gut metagenomic studies. Sci Rep 6: 26276.
-
28.
Holland JL, Louie L, Simor AE, Louie M, 2000. PCR detection of Escherichia coli O157:H7 directly from stools: evaluation of commercial extraction methods for purifying fecal DNA. J Clin Microbiol 38: 4108–4113.
-
29.
Oikarinen S, Tauriainen S, Viskari H, Simell O, Knip M, Virtanen S, Hyöty H, 2009. PCR inhibition in stool samples in relation to age of infants. J Clin Virol 44: 211–214.
-
30.
Widjojoatmodjo MN, Fluit AC, Torensma R, Verdonk GP, Verhoef J, 1992. The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples. J Clin Microbiol 30: 3195–3199.
-
31.
Liu J et al. 2013. A laboratory-developed TaqMan array card for simultaneous detection of 19 enteropathogens. J Clin Microbiol 51: 472–480.
-
32.
Nichols RA, Smith HV, 2004. Optimization of DNA extraction and molecular detection of Cryptosporidium oocysts in natural mineral water sources. J Food Prot 67: 524–532.
-
33.
Robertson LJ, Campbell AT, Smith HV, 1992. Survival of Cryptosporidium parvum oocysts under various environmental pressures. Appl Environ Microbiol 58: 3494–3500.
-
34.
Kaucner C, Stinear T, 1998. Sensitive and rapid detection of viable Giardia cysts and Cryptosporidium parvum oocysts in large-volume water samples with wound fiberglass cartridge filters and reverse transcription-PCR. Appl Environ Microbiol 64: 1743–1749.
-
35.
Mero S, Kirveskari J, Antikainen J, Ursing J, Rombo L, Kofoed PE, Kantele A, 2017. Multiplex PCR detection of Cryptosporidium sp, Giardia lamblia and Entamoeba histolytica directly from dried stool samples from Guinea-Bissauan children with diarrhoea. Infect Dis 49: 655–663.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 782 | 685 | 66 |
| Full Text Views | 579 | 12 | 2 |
| PDF Downloads | 238 | 12 | 2 |