Prevalence and diversity of enteric pathogens among cholera treatment centre patients with acute diarrhea in Uvira, Democratic Republic of Congo

Camille Williams, Oliver Cumming, Lynn Grignard, Baron Bashige Rumedeka, Jaime Mufitini Saidi, Daniel Grint, Chris Drakeley, Aurelie Jeandron, Camille Williams, Oliver Cumming, Lynn Grignard, Baron Bashige Rumedeka, Jaime Mufitini Saidi, Daniel Grint, Chris Drakeley, Aurelie Jeandron

Abstract

Background: Cholera remains a major global health challenge. Uvira, in the Democratic Republic of the Congo (DRC), has had endemic cholera since the 1970's and has been implicated as a possible point of origin for national outbreaks. A previous study among this population, reported a case confirmation rate of 40% by rapid diagnostic test (RDT) among patients at the Uvira Cholera Treatment Centre (CTC). This study considers the prevalence and diversity of 15 enteric pathogens in suspected cholera cases seeking treatment at the Uvira CTC.

Methods: We used the Luminex xTAG® multiplex PCR to test for 15 enteric pathogens, including toxigenic strains of V. cholerae in rectal swabs preserved on Whatman FTA Elute cards. Results were interpreted on MAGPIX® and analyzed on the xTAG® Data Analysis Software. Prevalence of enteric pathogens were calculated and pathogen diversity was modelled with a Poisson regression.

Results: Among 269 enrolled CTC patients, PCR detected the presence of toxigenic Vibrio cholerae in 38% (103/269) of the patients, which were considered to be cholera cases. These strains were detected as the sole pathogen in 36% (37/103) of these cases. Almost half (45%) of all study participants carried multiple enteric pathogens (two or more). Enterotoxigenic Escherichia coli (36%) and Cryptosporidium (28%) were the other most common pathogens identified amongst all participants. No pathogen was detected in 16.4% of study participants. Mean number of pathogens was highest amongst boys and girls aged 1-15 years and lowest in women aged 16-81 years. Ninety-three percent of toxigenic V. cholerae strains detected by PCR were found in patients having tested positive for V. cholerae O1 by RDT.

Conclusions: Our study supports previous results from DRC and other cholera endemic areas in sub-Sahara Africa with less than half of CTC admissions positive for cholera by PCR. More research is required to determine the causes of severe acute diarrhea in these low-resource, endemic areas to optimize treatment measures.

Trial registration: This study is part of the impact evaluation study entitled: "Impact Evaluation of Urban Water Supply Improvements on Cholera and Other Diarrheal Diseases in Uvira, Democratic Republic of Congo" registered on 10 October 2016 at clinicaltrials.gov Identification number: NCT02928341 .

Keywords: Cholera; Diarrhea; Endemic; Enteric pathogens; Sub-Sahara Africa.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Frequency of specific enteric pathogens detection among enrolled CTC patients

References

    1. Troeger C, Blacker BF, Khalil IA, Rao PC, Cao S, Zimsen SR, et al. Estimates of the global, regional, and national morbidity, mortality, and aetiologies of diarrhoea in 195 countries: a systematic analysis for the Global Burden of Disease Study 2016. Lancet Infect Dis. 2018;18:1211–1228. doi: 10.1016/S1473-3099(18)30362-1.
    1. Kotloff KL, Nataro JP, Blackwelder WC, Nasrin D, Farag TH, Panchalingam S, et al. 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. 2013;382:209–222. doi: 10.1016/S0140-6736(13)60844-2.
    1. Amour C, Gratz J, Mduma ER, Svensen E, Rogawski ET, McGrath M, et al. Epidemiology and Impact of Campylobacter Infection in Children in 8 Low-Resource Settings: Results from the MAL-ED Study. Clin Infect Dis. 2016;63(9):1171–1179.
    1. MAL-ED Network Investigators Early childhood cognitive development is affected by interactions among illness, diet, enteropathogens and the home environment: findings from the MAL-ED birth cohort study. BMJ Glob Health. 2018;3:752.
    1. Wagner EG, Lanoix J. Excreta disposal for rural areas and small communities. Monogr Ser. 1958;39:1–182.
    1. Federspiel F, Ali M. The cholera outbreak in Yemen: lessons learned and way forward. BMC Public Health. 2018;18(1338) Available from: 10.1186/s12889-018-6227-6. [cited 2019 15 Jan].
    1. Ali M, Nelson AR, Lena Lopez A, Sack DA. Updated Global Burden of Cholera in Endemic Countries. PLoS Negl Trop Dis. 2015;9(6) Available from: [cited 2018 5 Aug].
    1. The Global Task Force on Cholera Control . Ending Cholera: A Global Roadmap to 2030. 2017.
    1. Hartley DM, Glenn Morris J, Jr, Smith DL. Hyperinfectivity: a critical element in the ability of V. cholerae to cause epidemics? PLoS Med. 2005;3(1):0063–0069. doi: 10.1371/journal.pmed.0030007.
    1. Nelson EJ, Harris JB, Glenn Morris J Jr, Calderwood SB, Camilli A. Cholera transmission: the host, pathogen and bacteriophage dynamic. Nat Rev Microbiol. 2009;7(10).
    1. Griffith DC, Kelly-Hope LA, Miller MA. Review of reported cholera outbreaks worldwide, 1995-2005. Am J Trop Med Hyg. 2006;75(5):973–977. doi: 10.4269/ajtmh.2006.75.973.
    1. Lessler J, Moore SM, Luquero FJ, Mckay HS, Grais R, Henkens M, et al. Articles Mapping the burden of cholera in sub-Saharan Africa and implications for control: an analysis of data across geographical scales. Lancet. 2018;391:1908–1915. doi: 10.1016/S0140-6736(17)33050-7.
    1. World Health Organization. Meeting of the Strategic Advisory Group of Experts on Immunization, October 2018-- Conclusions and recommendations. Geneva; 2018. Available from: ,93,661-680No49 [cited 2019 21 Feb].
    1. Ciglenecki I, Azman AS, Jamet C, Serafini M, Luquero FJ, Cabrol J-C. Progress and challenges in using Oral cholera vaccines to control outbreaks: The Medicins sans Frontieres experience. JID. 2018;218(3):165–166. doi: 10.1093/infdis/jiy487.
    1. von Seidlein L, Deen JL. Preventing cholera outbreaks through early targeted interventions. PLoS Med. 2018;15(2):e1002510. doi: 10.1371/journal.pmed.1002510.
    1. Impact Evaluation of Urban Water Supply Improvements on Cholera and Other Diarrhoeal Diseases in Uvira, Democratic Republic of Congo Identifier: [Internet]. NIH Clinical Trials. 2016 [cited 2019 Feb 15]. Available from: .
    1. Bompangue D, Giraudoux P, Piarroux M, Mutombo G, Shamavu R, Sudre B, et al. Cholera Epidemics, War and Disasters around Goma and Lake Kivu: An Eight-Year Survey. PLoS Negl Trop Dis. 2009;3(5).
    1. Rebaudet S, Sudre B, Faucher B, Piarroux R. Environmental determinants of Cholera Outbreaks in Inland Africa: A Systematic Review of Main Transmission Foci and Propagation Routes. J Infect Dis. 2013;208(S1):S46–S54. doi: 10.1093/infdis/jit195.
    1. Jeandron A, Cumming O, Rumedeka BB, Saidi JM, Cousens S. Confirmation of cholera by rapid diagnostic test amongst patients admitted to the cholera treatment Centre in Uvira, Democratic Republic of the Congo. PLoS One. 2018;13(8) Available from: 10.1371/journal.pone.0201306.
    1. Bhuiyan NA, Qadri F, Faruque ASG, Malek MA, Salam MA, Nato F, et al. Use of Dipsticks for Rapid Diagnosis of Cholera Caused by Vibrio cholerae O1 and O139 from Rectal Swabs. J Clin Microbiol. 2003;41(8):3939–3941. doi: 10.1128/JCM.41.8.3939-3941.2003.
    1. Lalani T, Tisdale MD, Maguire JD, Wongsrichanalai C, Riddle MS, Tribble DR. 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. 2015;83(1):18–20. doi: 10.1016/j.diagmicrobio.2015.05.011.
    1. Luminex Molecular Diagnostics Inc. IVD xTAG ® gastrointestinal pathogen panel MLD-032-KPI-002 rev H. Toronto, ON, Canada; 2017. Available from: .
    1. Khare R, Espy MJ, Cebelinski E, Boxrud D, Sloan LM, Cunningham SA, et al. Comparative evaluation of two commercial multiplex panels for detection of gastrointestinal pathogens by use of clinical stool specimens. J Clin Microbiol. 2014;52(10):3667–3673. doi: 10.1128/JCM.01637-14.
    1. Perry MD, Corden SA, Howe RA. Evaluation of the Luminex xTAG gastrointestinal pathogen panel and the Savyon diagnostics gastrointestinal infection panel for the detection of enteric pathogens in clinical samples. J Med Microbiol. 2014;63:1419–1426. doi: 10.1099/jmm.0.074773-0.
    1. Navidad JF, Griswold DJ, Gradus MS, Bhattacharyya S. Evaluation of Luminex xTAG Gastrointestinal Pathogen Analyte-Specific Reagents for High-Throughput, Simultaneous Detection of Bacteria, Viruses, and Parasites of Clinical and Public Health Importance. J Clin Microbiol. 2013;51(9):3018–3024. doi: 10.1128/JCM.00896-13.
    1. Becker SL, Chatigre JK, Gohou J-P, Coulibaly JT, Leuppi R, Polman K, et al. Combined stool-based multiplex PCR and microscopy for enhanced pathogen detection in patients with persistent diarrhoea and asymptomatic controls from Côte d’Ivoire. Clin Microbiol Infect. 2015;21:591.e1–591.e10. doi: 10.1016/j.cmi.2015.02.016.
    1. Eibach D, Krumkamp R, Hahn A, Sarpong N, Adu-Sarkodie Y, Leva A, et al. Application of a multiplex PCR assay for the detection of gastrointestinal pathogens in a rural African setting. BMC Infect Dis. 2016;16(1):150. doi: 10.1186/s12879-016-1481-7.
    1. GE Life Sciences. Whatman FTA Elute Data file 28–9844-02 AA. Buckinhamshire, UK; 2011. p. 2–5.
    1. Cameron AC, Trivedi PK. Microeconomics using Stata. Revised Ed. College Station, Texas: Stata Press; 2010. pp. 572–576.
    1. Tribble DR. Antibiotic Therapy for Acute Watery Diarrhea and Dysentery. Mil Med. 2017;182 Available from: . Cited 2019 Feb 17.
    1. Sauvageot D, Njanpop-Lafourcade B-M, Akilimali L, Anne J-C, Bidjada P, Bompangue D, et al. Cholera incidence and mortality in sub-Saharan African sites during multi-country surveillance. PLOS Neglected Top Dis. 2016;10(5):1–16.
    1. Dorman MJ, Domman D, Uddin MI, Sharmin S, Afrad MH, Begum YA, et al. High quality reference genomes for toxigenic and non-toxigenic Vibrio cholerae serogroup O139. Available from: 10.1038/s41598-019-41883-x [cited 2020 18 Aug].
    1. Tobin-D M, Smith AR, Bulens SN, Thomas S, Hodel M, Izumiya H, et al. Severe Diarrhea Caused by Cholera Toxin-Producing Vibrio cholerae Serogroup O75 Infections Acquired in the Southeastern United States. 2008;1035. Available from: . Cited 2020 Aug 18.
    1. Liu J, Platts-Mills JA, Juma J, Kabir F, Nkeze J, Okoi C, et al. Use of quantitative molecular diagnostic methods to identify causes of diarrhoea in children: a reanalysis of the GEMS case-control study. Lancet. 2016;388:1291–1301. doi: 10.1016/S0140-6736(16)31529-X.
    1. Harris AM, Chowdhury F, Begum YA, Khan AI, Faruque ASG, Svennerholm A-M, et al. Shifting Prevalence of Major Diarrheal Pathogens in Patients Seeking Hospital Care during Floods in in Dhaka, Bangladesh. Am J Trop Med Hyg. 2008;79(5):708–714. doi: 10.4269/ajtmh.2008.79.708.
    1. Andersson M, Kabayiza J-C, Elfving K, Nilsson S, Msellem MI, Artensson AM, et al. Coinfection with Enteric Pathogens in East African Children with Acute Gastroenteritis-Associations and Interpretations. Am J Trop Med Hyg. 2018;98(6):1566–1570. doi: 10.4269/ajtmh.17-0473.
    1. Levine MM, Robins-Browne RM. Factors That Explain Excretion of Enteric Pathogens by Persons Without Diarrhea. Clin Infect Dis. 2012;55(4):303–311. doi: 10.1093/cid/cis789.
    1. Platts-Mills JA, Liu J, Rogawski ET, Kabir F, Lertsethtakarn P, Siguas M, et al. Use of quantitative molecular diagnostic methods to assess the aetiology, burden, and clinical characteristics of diarrhoea in children in low-resource settings: a reanalysis of the MAL-ED cohort study. Lancet Glob Health. 2018;6(12):e1309–e1318. doi: 10.1016/S2214-109X(18)30349-8.
    1. Kosek MN, Ahmed T, Bhutta Z, Caulfield L, Guerrant R, Houpt E, et al. Causal Pathways from Enteropathogens to Environmental Enteropathy: Findings from the MAL-ED Birth Cohort Study. EBioMedicine. 2017;18:109–117. doi: 10.1016/j.ebiom.2017.02.024.
    1. Cairncross S, Feachem SR. Earthscan Water Text. Third. London: Routledge; 2018. Environmental Health Engineering in the Tropics: Water, Sanitation and Disease Control; pp. 4–9.
    1. Schaetti C, Sundaram N, Merten S, Ali SM, Nyambedha EO, Lapika B, et al. Comparing sociocultural features of cholera in three endemic African settings. BMC Med. 2013;11:1. doi: 10.1186/1741-7015-11-206.
    1. Kabayiza J-C, Andersson ME, Welinder-Olsson C, Bergström T, Muhirwa G, Lindh M. Comparison of rectal swabs and faeces for real-time PCR detection of enteric agents in Rwandan children with gastroenteritis. BMC Infect Dis. 2013;13(447) Available from: [cited 2019 14 Feb].
    1. Lalani T, Tisdale MD, Liu J, Mitra I, Philip C, Odundo E, et al. Comparison of stool collection and storage on Whatman FTA® Elute cards versus frozen stool for enteropathogen detection using the TaqMan® Array Card PCR assay. PLoS One. 2018;13(8):e0202178. doi: 10.1371/journal.pone.0202178.
    1. Lau H, Hurt AC. Assessment of the RNASound RNA Sampling Card for the Preservation of Influenza Virus RNA. Front Microbiol. 2016;7(1736) Available from: [cited 2019 18 Feb].
    1. Jones S, Sutherland CJ, Hermsen C, Arens T, Teelen K, Hallett R, et al. Filter paper collection of Plasmodium falciparum mRNA for detecting low-density gametocytes. Malar J. 2012;11(266) Available from: [cited 2019 14 Mar].
    1. Rogawski ET, Platts-Mills JA, Liu J, Operario DJ, Taniuchi M, Gratz JM, et al. Use of quantitative molecular diagnostic methods to investigate the effect of enteropathogen infections on linear growth in children in low-resource settings: longitudinal analysis of results from the MAL-ED cohort study. Lancet Glob Health. 2018;6:1319–1347. doi: 10.1016/S2214-109X(18)30351-6.
    1. Liu J, Gratz J, Amour C, Nshama R, Walongo T, Maro A, et al. Optimization of quantitative PCR methods for Enteropathogen detection. PLoS One. 2016;11(6):e0158199. doi: 10.1371/journal.pone.0158199.
    1. Platts-Mills JA, Gratz J, Mduma E, Svensen E, Amour C, Liu J, et al. Association Between Stool Enteropathogen Quantity and Disease in Tanzanian Children Using TaqMan Array Cards: A Nested Case-Control Study. Am J Trop Med Hyg. 2014;90(1):133–138. doi: 10.4269/ajtmh.13-0439.
    1. Kang G, Iturriza-Gomara M, Wheeler JG, Crystal P, Monica B, Ramani S, et al. Quantitation of Group A Rotavirus by Real-Time Reverse-Transcription-Polymerase Chain Reaction: Correlation With Clinical Severity in Children in South India. J Med Virol. 2004;74(1):118–122. doi: 10.1002/jmv.20053.

Source: PubMed

3
订阅