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. 2014 Nov 1;59 Suppl 4(Suppl 4):S225-32.
doi: 10.1093/cid/ciu413.

Microbiologic methods utilized in the MAL-ED cohort study

Eric Houpt  1 , Jean Gratz  1 , Margaret Kosek  2 , Anita K M Zaidi  3 , Shahida Qureshi  3 , Gagandeep Kang  4 , Sudhir Babji  4 , Carl Mason  5 , Ladaporn Bodhidatta  5 , Amidou Samie  6 , Pascal Bessong  6 , Leah Barrett  1 , Aldo Lima  7 , Alexandre Havt  7 , Rashidul Haque  8 , Dinesh Mondal  8 , Mami Taniuchi  1 , Suzanne Stroup  1 , Monica McGrath  9 , Dennis Lang  10 ; MAL-ED Network Investigators
Collaborators, Affiliations

Microbiologic methods utilized in the MAL-ED cohort study

Eric Houpt et al. Clin Infect Dis. .

Abstract

A central hypothesis of The Etiology, Risk Factors and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development (MAL-ED) study is that enteropathogens contribute to growth faltering. To examine this question, the MAL-ED network of investigators set out to achieve 3 goals: (1) develop harmonized protocols to test for a diverse range of enteropathogens, (2) provide quality-assured and comparable results from 8 global sites, and (3) achieve maximum laboratory throughput and minimum cost. This paper describes the rationale for the microbiologic assays chosen and methodologies used to accomplish the 3 goals.

Keywords: ELISA; PCR; culture; enteropathogen; microscopy.

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Figures

Figure 1.
Figure 1.
Workflow for stool specimen from collection to assay. Abbreviations: ELISA, enzyme-linked immunosorbent assay; O&P, ova and parasites; PCR, polymerase chain reaction.
Figure 2.
Figure 2.
Multiplex polymerase chain reaction (PCR) gel electrophoresis assay for diarrheagenic Escherichia coli. The E. coli isolates underwent DNA extraction and multiplex PCR for virulence genes as described in the Methods. Virulence genes are discriminated by band length. Abbreviations: EAEC, enteroaggregative Escherichia coli; EHEC, enterohemorrhagic Escherichia coli; EIEC, enteroinvasive Escherichia coli; EPEC, enteropathogenic Escherichia coli; ETEC, enterotoxigenic Escherichia coli; nfw, nuclease free water.
See this image and copyright information in PMC

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