title: Diagnostic microbiologic methods in the GEMS-1 case/control study.
abstract: To understand the etiology of moderate-to-severe diarrhea among children in high mortality areas of sub-Saharan Africa and South Asia, we performed a comprehensive case/control study of children aged <5 years at 7 sites. Each site employed an identical case/control study design and each utilized a uniform comprehensive set of microbiological assays to identify the likely bacterial, viral and protozoal etiologies. The selected assays effected a balanced consideration of cost, robustness and performance, and all assays were performed at the study sites. Identification of bacterial pathogens employed streamlined conventional bacteriologic biochemical and serological algorithms. Diarrheagenic Escherichia coli were identified by application of a multiplex polymerase chain reaction assay for enterotoxigenic, enteroaggregative, and enteropathogenic E. coli. Rotavirus, adenovirus, Entamoeba histolytica, Giardia enterica, and Cryptosporidium species were detected by commercially available enzyme immunoassays on stool samples. Samples positive for adenovirus were further evaluated for adenovirus serotypes 40 and 41. We developed a novel multiplex assay to detect norovirus (types 1 and 2), astrovirus, and sapovirus. The portfolio of diagnostic assays used in the GEMS study can be broadly applied in developing countries seeking robust cost-effective methods for enteric pathogen detection.
authors.affiliation: Center for Vaccine Development, University of Maryland School of Medicine, Baltimore, MD, USA.
authors.forename: Sandra
authors.initials: S
authors.lastname: Panchalingam
authors.forename: Martin
authors.initials: M
authors.lastname: Antonio
authors.forename: Anowar
authors.initials: A
authors.lastname: Hossain
authors.forename: Inacio
authors.initials: I
authors.lastname: Mandomando
authors.forename: Ben
authors.initials: B
authors.lastname: Ochieng
authors.forename: Joseph
authors.initials: J
authors.lastname: Oundo
authors.forename: T
authors.initials: T
authors.lastname: Ramamurthy
authors.forename: Boubou
authors.initials: B
authors.lastname: Tamboura
authors.forename: Anita K M
authors.initials: AK
authors.lastname: Zaidi
authors.forename: William
authors.initials: W
authors.lastname: Petri
authors.forename: Eric
authors.initials: E
authors.lastname: Houpt
authors.forename: Patrick
authors.initials: P
authors.lastname: Murray
authors.forename: Valeria
authors.initials: V
authors.lastname: Prado
authors.forename: Roberto
authors.initials: R
authors.lastname: Vidal
authors.forename: Duncan
authors.initials: D
authors.lastname: Steele
authors.forename: Nancy
authors.initials: N
authors.lastname: Strockbine
authors.forename: Philippe
authors.initials: P
authors.lastname: Sansonetti
authors.forename: Roger I
authors.initials: RI
authors.lastname: Glass
authors.forename: Roy M
authors.initials: RM
authors.lastname: Robins-Browne
authors.forename: Marija
authors.initials: M
authors.lastname: Tauschek
authors.forename: Ann-Marie
authors.initials: AM
authors.lastname: Svennerholm
authors.forename: Lynette Y
authors.initials: LY
authors.lastname: Berkeley
authors.forename: Karen
authors.initials: K
authors.lastname: Kotloff
authors.forename: Myron M
authors.initials: MM
authors.lastname: Levine
authors.forename: James P
authors.initials: JP
authors.lastname: Nataro
mesh_terms: D017954:Africa South of the Sahara; D001211:Asia, Western; D016022:Case-Control Studies; D003458:Cryptosporidium / Q000302:isolation & purification; D003967:Diarrhea / Q000209:etiology / Q000382:microbiology* / Q000469:parasitology* / Q000821:virology; D004748:Entamoeba histolytica / Q000302:isolation & purification; D004926:Escherichia coli / Q000302:isolation & purification; D005872:Giardia / Q000302:isolation & purification; D006801:Humans; D007124:Immunoenzyme Techniques; D008828:Microbiological Techniques / Q000379:methods; D015337:Multicenter Studies as Topic / Q000379:methods; D010274:Parasitology / Q000379:methods; D016133:Polymerase Chain Reaction; D011785:Quality Assurance, Health Care; D011786:Quality Control; D014773:Virology / Q000379:methods; D014780:Viruses / Q000302:isolation & purification