Volume 93, Issue 5
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645



Molecular identification of the invasion plasmid antigen-H () gene has been established as a useful detection mechanism for spp. The Global Enteric Multicenter Study (GEMS) identified the etiology and burden of moderate-to-severe diarrhea (MSD) in sub-Saharan Africa and south Asia using a case–control study and traditional culture techniques. Here, we used quantitative polymerase chain reaction (qPCR) to identify spp. in 2,611 stool specimens from GEMS and compared these results to those using culture. Demographic and nutritional characteristics were assessed as possible risk factors. The qPCR identified more cases of shigellosis than culture; however, the distribution of demographic characteristics was similar by both methods. In regression models adjusting for quantity, age, and site, children who were exclusively breast-fed had significantly lower odds of MSD compared with children who were not breast-fed (odds ratio [OR] = 0.47, 95% confidence interval (CI) = 0.28–0.81). The association between quantity and MSD increased with age, with a peak in children of 24–35 months of age (OR = 8.2, 95% CI = 4.3–15.7) and the relationship between quantity and disease was greatest in Bangladesh (OR = 13.2, 95% CI = 7.3–23.8). This study found that qPCR identified more cases of and age, site, and breast-feeding status were significant risk factors for MSD.


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  1. Kotloff KL, Nataro JP, Blackwelder WC, Nasrin D, Farag TH, Panchalingam S, Wu Y, Sow SO, Sur D, Breiman RF, Faruque AS, Zaidi AK, Saha D, Alonso PL, Tamboura B, Sanogo D, Onwuchekwa U, Manna B, Ramamurthy T, Kanungo S, Ochieng JB, Omore R, Oundo JO, Hossain A, Das SK, Ahmed S, Qureshi S, Quadri F, Adegbola RA, Antonio M, Hossain MJ, Akinsola A, Mandomando I, Nhampossa T, Acácio S, Biswas K, O'Reilly CE, Mintz ED, Berkeley LY, Muhsen K, Sommerfelt H, Robins-Browne RM, Levine MM, , 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: 209222.[Crossref]
  2. Barletta F, Ochoa TJ, Mercado E, Ruiz J, Ecker L, Lopez G, Mispireta M, Gil AI, Lanata CF, Cleary TG, , 2011. Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections. Clin Infect Dis 53: 12231229.[Crossref]
  3. Dutta S, Chatterjee A, Dutta P, Rajendran K, Roy S, Pramanik KC, Bhattacharya SK, , 2001. Sensitivity and performance characteristics of a direct PCR with stool samples in comparison to conventional techniques for diagnosis of Shigella and enteroinvasive Escherichia coli infection in children with acute diarrhea in Calcutta, India. J Med Microbiol 50: 667674.[Crossref]
  4. Barletta F, Mercado EH, Lluque A, Ruiz J, Cleary TG, Ochoa TJ, , 2013. Multiplex real-time PCR for detection of Campylobacter, Salmonella, and Shigella . J Clin Microbiol 51: 28222829.[Crossref]
  5. Taniuchi M, Walters CC, Gratz J, Maro A, Kumburu H, Serichantalergs O, Sethabutr O, Bodhidatta L, Kibiki G, Toney DM, Berkeley L, Nataro JP, Houpt ER, , 2012. Development of a multiplex polymerase chain reaction assay for diarrheagenic Escherichia coli and Shigella spp. and its evaluation on colonies, culture broths, and stool. Diagn Microbiol Infect Dis 73: 121128.[Crossref]
  6. Von Seidlein L, Kim DR, Ali M, Lee H, Wang X, Thiem VD, Canh DG, Chaicumpa W, Agtini MD, Hossain A, Bhutta ZA, Mason C, Sethabutr O, Talukder K, Nair GB, Deen JL, Kotloff K, Clemens J, , 2006. A multicentre study of Shigella diarrhoea in six Asian countries: disease burden, clinical manifestations, and microbiology. PLoS Med 3: e353.[Crossref]
  7. Samosornsuk S, Chaicumpa W, von Seidlein L, Clemens JD, Sethabutr O, , 2007. Using real-time PCR to detect shigellosis: ipaH detection in Kaeng-Khoi District, Saraburi Province, Thailand. Thammasat Int J Sci Technol 12: 5257.
  8. Vu DT, Sethabutr O, Von Seidlein L, Tran VT, Do GC, Bui TC, Le HT, Lee H, Houng H-S, Hale TL, Clemens JD, Mason C, Dang DT, , 2004. Detection of Shigella by a PCR assay targeting the ipaH gene suggests increased prevalence of shigellosis in Nha Trang, Vietnam. J Clin Microbiol 42: 20312035.[Crossref]
  9. Lindsay B, Ochieng JB, Ikumapayi UN, Toure A, Ahmed D, Li S, Panchalingam S, Levine MM, Kotloff K, Rasko DA, Morris CR, Juma J, Fields BS, Dione M, Malle D, Becker SM, Houpt ER, Nataro JP, Sommerfelt H, Pop M, Oundo J, Antonio M, Hossain A, Tamboura B, Stine OC, , 2013. Quantitative PCR for detection of Shigella improves ascertainment of Shigella burden in children with moderate-to-severe diarrhea in low-income countries. J Clin Microbiol 51: 17401746.[Crossref]
  10. Lindsay B, Pop M, Antonio M, Walker AW, Mai V, Ahmed D, Oundo J, Tamboura B, Panchalingam S, Levine MM, Kotloff K, Li S, Magder LS, Paulson JN, Liu B, Ikumapayi U, Ebruke C, Dione M, Adeyemi M, Rance R, Stares MD, Ukhanova M, Barnes B, Lewis I, Ahmed F, Alam MT, Amin R, Siddiqui S, Ochieng JB, Ouma E, Juma J, Mailu E, Omore R, O'Reilly CE, Hannis J, Manalili S, Deleon J, Yasuda I, Blyn L, Ranken R, Li F, Housley R, Ecker DJ, Hossain MA, Breiman RF, Morris JG, McDaniel TK, Parkhill J, Saha D, Sampath R, Stine OC, Nataro JP, , 2013. Survey of culture, goldengate assay, universal biosensor assay, and 16S rRNA gene sequencing as alternative methods of bacterial pathogen detection. J Clin Microbiol 51: 32633269.[Crossref]
  11. Nair GB, Ramamurthy T, Bhattacharya MK, Krishnan T, Ganguly S, Saha DR, Rajendran K, Manna B, Ghosh M, Okamoto K, Takeda Y, , 2010. Emerging trends in the etiology of enteric pathogens as evidenced from an active surveillance of hospitalized diarrheal patients in Kolkata, India. Gut Pathog 2: 4.[Crossref]
  12. Lindsay B, Ramamurthy T, Sen Gupta S, Takeda Y, Rajendran K, Nair GB, Stine OC, , 2011. Diarrheagenic pathogens in polymicrobial infections. Emerg Infect Dis 17: 606611.[Crossref]
  13. Lindsay B, Oundo J, Hossain MA, Antonio M, Tamboura B, Walker AW, Paulson JN, Parkhill J, Omore R, Faruque AS, Das SK, Ikumapayi UN, Adeyemi M, Sanogo D, Saha D, Sow S, Farag TH, Nasrin D, Li S, Panchalingam S, Levine MM, Kotloff K, Magder LS, Hungerford L, Sommerfelt H, Pop M, Nataro JP, Stine OC, , 2015. Microbiota that affect risk for shigellosis in children in low-income countries. Emerg Infect Dis 21: 242250.[Crossref]
  14. Kotloff KL, Blackwelder WC, Nasrin D, Nataro JP, Farag TH, van Eijk A, Adegbola RA, Alonso PL, Breiman RF, Faruque ASG, Saha D, Sow SO, Sur D, Zaidi AKM, Biswas K, Panchalingam S, Clemens JD, Cohen D, Glass RI, Mintz ED, Sommerfelt H, Levine MM, , 2012. The Global Enteric Multicenter Study (GEMS) of diarrheal disease in infants and young children in developing countries: epidemiologic and clinical methods of the case/control study. Clin Infect Dis 55 (Suppl 4): S232S245.[Crossref]
  15. Panchalingam S, Antonio M, Hossain A, Mandomando I, Ochieng B, Oundo J, Ramamurthy T, Tamboura B, Zaidi AKM, Petri W, Houpt E, Murray P, Prado V, Vidal R, Steele D, Strockbine N, Sansonetti P, Glass RI, Robins-Browne RM, Tauschek M, Svennerholm A-M, Berkeley LY, Kotloff K, Levine MM, Nataro JP, , 2012. Diagnostic microbiologic methods in the GEMS-1 case/control study. Clin Infect Dis 55 (Suppl 4): S294S302.[Crossref]
  16. Taniuchi M, Sobuz SU, Begum S, Platts-Mills JA, Liu J, Yang Z, Wang X-Q, Petri WA, Jr Haque R, Houpt ER, , 2013. Etiology of diarrhea in Bangladeshi infants in the first year of life analyzed using molecular methods. J Infect Dis 208: 17941802.[Crossref]
  17. Clemens JD, Stanton B, Stoll B, Shahid NS, Banu H, Chowdhury AK, , 1986. Breast feeding as a determinant of severity in shigellosis. Evidence for protection throughout the first three years of life in Bangladeshi children. Am J Epidemiol 123: 710720.[Crossref]
  18. Ahmed F, Clemens JD, Rao MR, Sack DA, Khan MR, Haque E, , 1992. Community-based evaluation of the effect of breast-feeding on the risk of microbiologically confirmed or clinically presumptive shigellosis in Bangladeshi children. Pediatrics 90: 406411.
  19. Ahmed F, Clemens JD, Rao MR, Ansaruzzaman M, Haque E, , 1997. Epidemiology of shigellosis among children exposed to cases of Shigella dysentery: a multivariate assessment. Am J Trop Med Hyg 56: 258264.
  20. UNAIDS, 2014. HIV Prevalence-Ages 15–49 (%). Available at: http://www.unaids.org/en/dataanalysis/datatools/aidsinfo.
  21. Ricci KA, Girosi F, Tarr PI, Lim Y-W, Mason C, Miller M, Hughes J, von Seidlein L, Agosti JM, Guerrant RL, , 2006. Reducing stunting among children: the potential contribution of diagnostics. Nature 444 (Suppl 1): 2938.

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  • Received : 22 May 2014
  • Accepted : 01 Jun 2015

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