Detection and Molecular Serotyping With a Customized TaqMan Array Card in the Enterics for Global Health (EFGH): Surveillance Study

Background: Quantitative polymerase chain reaction (qPCR) targeting has been proven to be highly efficient in detecting in clinical samples compared to culture-based methods, which underestimate burden by 2- to 3-fold. qPCR assays have also been developed for speciation and serotyping, which is critical for both vaccine development and evaluation.

Methods: The Enterics for Global Health (EFGH) surveillance study will utilize a customized real-time PCR-based TaqMan Array Card (TAC) interrogating 82 targets, for the detection and differentiation of spp, , 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 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 -attributed disease burden, which is critical to informing policy and in the design of future clinical trials.

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