Volume 94, Issue 3
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



Biological confirmation of the presence of in clinical and environmental samples is often constrained due to resource- and labor-intensive gold standard methods. To develop low-cost, simple, and sustainable surveillance techniques, we modified previously published specimen sampling and culture techniques and applied the use of enriched dipstick testing in conjunction with the use of filter paper for DNA specimen preservation during clinical and environmental surveillance in the Far North of Cameroon from August 2013 to October 2014. The enriched dipstick methodology during routine use in a remote setting demonstrated a specificity of 99.8% compared with polymerase chain reaction (PCR). The novel application of filter paper as a preservation method for cholera DNA specimens reduced the need for cold chain storage and allowed for PCR characterization and confirmation of . The application of basic technologies such as the enriched dipstick, the use of simplified gauze filtration for environmental sample collection, and the use of filter paper for sample preservation enabled early case identification with reduced logistics and supply cost while reporting minimal false-positive results. Simplified laboratory and epidemiological methodologies can improve the feasibility of cholera surveillance in rural and resource-constrained areas, facilitating early case detection and rapid response implementation.

[open-access] This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


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  1. Griffith DC, Kelly-Hope LA, Miller MA, , 2006. Review of reported cholera outbreaks worldwide, 1995–2005. Am J Trop Med Hyg 75: 973977.
  2. World Health Organization, 2014. Cholera, 2013. Wkly Epidemiol Rec 89: 345355.
  3. Hampton LM, Zell ER, Schrag S, Cohen AL, , 2012. Sentinel versus population-based surveillance of pneumococcal conjugate vaccine effectiveness. Bull World Health Organ 90: 568577.[Crossref]
  4. Sack RB, Siddique AK, Longini IM, Jr Nizam A, Yunus M, Islam MS, Morris JG, Jr Ali A, Huq A, Nair GB, Qadri F, Faruque SM, Sack DA, Colwell RR, , 2003. A 4-year study of the epidemiology of Vibrio cholerae in four rural areas of Bangladesh. J Infect Dis 187: 96101.[Crossref]
  5. Deen JL, von Seidlein L, Sur D, Agtini M, Lucas ME, Lopez AL, Kim DR, Ali M, Clemens JD, , 2008. The high burden of cholera in children: comparison of incidence from endemic areas in Asia and Africa. PLoS Negl Trop Dis 2: e173.[Crossref]
  6. World Health Organization, 1993. Guidelines for Cholera Control. Geneva, Switzerland: WHO.
  7. Harris JR, Cavallaro EC, de Nobrega AA, Dos SBJC, Bopp C, Parsons MB, Djalo D, Fonseca FG, Ba U, Semedo A, Sobel J, Mintz ED, , 2009. Field evaluation of Crystal VC® Rapid Dipstick test for cholera during a cholera outbreak in Guinea-Bissau. Trop Med Int Health 14: 11171121.[Crossref]
  8. Ley B, Khatib AM, Thriemer K, von Seidlein L, Deen J, Mukhopadyay A, Chang NY, Hashim R, Schmied W, Busch CJ, Reyburn R, Wierzba T, Clemens JD, Wilfing H, Enwere G, Aguado T, Jiddawi MS, Sack D, Ali SM, , 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.[Crossref]
  9. Mukherjee P, Ghosh S, Ramamurthy T, Bhattacharya MK, Nandy RK, Takeda Y, Nair GB, Mukhopadhyay AK, , 2010. Evaluation of a rapid immunochromatographic dipstick kit for diagnosis of cholera emphasizes its outbreak utility. Jpn J Infect Dis 63: 234238.
  10. George CM, Rashid MU, Sack DA, Sack RB, Saif-Ur-Rahman KM, Azman AS, Monira S, Bhuyian SI, Zillur Rahman KM, Toslim Mahmud M, Mustafiz M, Alam M, , 2014. Evaluation of enrichment method for the detection of Vibrio cholerae O1 using a rapid dipstick test in Bangladesh. Trop Med Int Health 19: 301307.[Crossref]
  11. Wang XY, Ansaruzzaman M, Vaz R, Mondlane C, Lucas ME, von Seidlein L, Deen JL, Ampuero S, Puri M, Park T, Nair GB, Clemens JD, Chaignat CL, Rajerison M, Nato F, Fournier JM, , 2006. Field evaluation of a rapid immunochromatographic dipstick test for the diagnosis of cholera in a high-risk population. BMC Infect Dis 6: 17.[Crossref]
  12. Bhuiyan NA, Qadri F, Faruque AS, Malek MA, Salam MA, Nato F, Fournier JM, Chanteau S, Sack DA, Balakrish Nair G, , 2003. Use of dipsticks for rapid diagnosis of cholera caused by Vibrio cholerae O1 and O139 from rectal swabs. J Clin Microbiol 41: 39393941.[Crossref]
  13. Chakraborty S, Alam M, Scobie HM, Sack DA, , 2013. Adaptation of a simple dipstick test for detection of Vibrio cholerae O1 and O139 in environmental water. Front Microbiol 4: 320.
  14. Ako AA, Shimada J, Eyong GE, Fantong WY, , 2010. Access to potable water and sanitation in Cameroon within the context of Millennium Development Goals (MDGS). Water Sci Technol 61: 13171339.[Crossref]
  15. International Federation of Red Cross and Red Crescent Societies I, 2011. Cameroon: Cholera in Northern Cameroon. Available at: http://reliefweb.int/report/cameroon/cholera-northern-cameroon-dref-operation-final-report-mdrcm009. Accessed December 15, 2015
  16. Debes A, Chakraborty S, Sack DA, , 2015. Manual for detecting Vibrio cholerae O1 from fecal samples using an enriched dipstick assay—a low-cost, simplified method of confirming cholera. Available at: https://www.stopcholera.org/sites/cholera/files/stop_cholera_fact_sheet_1_5_revised.pdf. Accessed October 20, 2015.
  17. Kain KC, Lanar DE, , 1991. Determination of genetic variation within Plasmodium falciparum by using enzymatically amplified DNA from filter paper disks impregnated with whole blood. J Clin Microbiol 29: 11711174.
  18. Bauer A, Rorvik LM, , 2007. A novel multiplex PCR for the identification of Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus . Lett Appl Microbiol 45: 371375.[Crossref]
  19. Nandi B, Nandy RK, Mukhopadhyay S, Nair GB, Shimada T, Ghose AC, , 2000. Rapid method for species-specific identification of Vibrio cholerae using primers targeted to the gene of outer membrane protein OmpW. J Clin Microbiol 38: 41454151.
  20. Hoshino K, Yamasaki S, Mukhopadhyay AK, Chakraborty S, Basu A, Bhattacharya SK, Nair GB, Shimada T, Takeda Y, , 1998. Development and evaluation of a multiplex PCR assay for rapid detection of toxigenic Vibrio cholerae O1 and O139. FEMS Immunol Med Microbiol 20: 201207.[Crossref]
  21. Hasan NA, Chowdhury WB, Rahim N, Sultana M, Shabnam SA, Mai V, Ali A, Morris GJ, Sack RB, Huq A, Colwell RR, Endtz HP, Cravioto A, Alam M, , 2010. Metagenomic 16S rDNA targeted PCR-DGGE in determining bacterial diversity in aquatic ecosystem. Bangladesh J Microbiol 27: 4650.
  22. StataCorp, 2014. STATA Data Analysis and Statistical Software. Available at: http://www.stata.com/.

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  • Received : 03 Jul 2015
  • Accepted : 14 Nov 2015

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