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Abstract
Background
Quantitative polymerase chain reaction (qPCR) targeting ipaH has been proven to be highly efficient in detecting Shigella in clinical samples compared to culture-based methods, which underestimate Shigella burden by 2- to 3-fold. qPCR assays have also been developed for Shigella speciation and serotyping, which is critical for both vaccine development and evaluation.
Methods
The Enterics for Global Health (EFGH) Shigella surveillance study will utilize a customized real-time PCR–based TaqMan Array Card (TAC) interrogating 82 targets, for the detection and differentiation of Shigella spp, Shigella sonnei, Shigella flexneri serotypes, other diarrhea-associated enteropathogens, and antimicrobial resistance (AMR) genes. Total nucleic acid will be extracted from rectal swabs or stool samples, and assayed on TAC. Quantitative analysis will be performed to determine the likely attribution of Shigella and other particular etiologies of diarrhea using the quantification cycle cutoffs derived from previous studies. The qPCR results will be compared to conventional culture, serotyping, and phenotypic susceptibility approaches in EFGH.
Conclusions
TAC enables simultaneous detection of diarrheal etiologies, the principal pathogen subtypes, and AMR genes. The high sensitivity of the assay enables more accurate estimation of Shigella-attributed disease burden, which is critical to informing policy and in the design of future clinical trials.
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Details
1 School of Public Health, Qingdao University , Qingdao , China
2 Asociación Benéfica PRISMA , Iquitos, Loreto , Peru
3 Infectious Diseases Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka , Bangladesh
4 Medical Research Council Unit The Gambia, London School of Hygiene and Tropical Medicine , Fajara , The Gambia
5 Centre pour le Développement des Vaccins du Mali (CVD-Mali), Bamako , Mali
6 Malawi Liverpool Wellcome Research Programme , Blantyre , Malawi
7 Division of Infectious Diseases and International Health, University of Virginia , Charlottesville, Virginia , USA
8 Department of Pediatrics and Child Health, The Aga Khan University , Karachi , Pakistan
9 Center for Global Health Research, Kenya Medical Research Institute , Kisumu , Kenya
10 Institute of Infection, Veterinary and Ecological Sciences, Department of Clinical Infection, Microbiology and Immunology, University of Liverpool , Liverpool , United Kingdom
11 Department of Global Health, University of Washington , Seattle, Washington , USA
12 Center for Vaccine Development and Global Health, University of Maryland School of Medicine , Baltimore, Maryland , USA