Taniuchi M., Walters C.C., Gratz J., Maro A., Kumburu H., Serichantalergs O., Sethabutr O., Bodhidatta L., Kibiki G., Toney D.M., Berkeley L., Nataro J.P., Houpt E.R.
Division of Infectious Diseases and International Health, University of Virginia, PO Box 801340, Charlottesville, VA 22908, United States; Division of Consolidated Laboratory Services, Virginia Department of General Services, Richmond, VA 23219, United States; Biotechnology Laboratory, Kilimanjaro Christian Medical Centre, PO Box 3010, Moshi, Tanzania; Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences, Phyathai, Bangkok 10400, Thailand; Center for Vaccine Development, University of Maryland, Baltimore, MD 21201, United States; Department of Pediatrics, University of Virginia, PO Box 800386, Charlottesville, VA 22908, United States
Taniuchi, M., Division of Infectious Diseases and International Health, University of Virginia, PO Box 801340, Charlottesville, VA 22908, United States; Walters, C.C., Division of Consolidated Laboratory Services, Virginia Department of General Services, Richmond, VA 23219, United States; Gratz, J., Division of Infectious Diseases and International Health, University of Virginia, PO Box 801340, Charlottesville, VA 22908, United States, Biotechnology Laboratory, Kilimanjaro Christian Medical Centre, PO Box 3010, Moshi, Tanzania; Maro, A., Biotechnology Laboratory, Kilimanjaro Christian Medical Centre, PO Box 3010, Moshi, Tanzania; Kumburu, H., Biotechnology Laboratory, Kilimanjaro Christian Medical Centre, PO Box 3010, Moshi, Tanzania; Serichantalergs, O., Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences, Phyathai, Bangkok 10400, Thailand; Sethabutr, O., Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences, Phyathai, Bangkok 10400, Thailand; Bodhidatta, L., Department of Enteric Diseases, Armed Forces Research Institute of Medical Sciences, Phyathai, Bangkok 10400, Thailand; Kibiki, G., Biotechnology Laboratory, Kilimanjaro Christian Medical Centre, PO Box 3010, Moshi, Tanzania; Toney, D.M., Division of Consolidated Laboratory Services, Virginia Department of General Services, Richmond, VA 23219, United States; Berkeley, L., Center for Vaccine Development, University of Maryland, Baltimore, MD 21201, United States; Nataro, J.P., Department of Pediatrics, University of Virginia, PO Box 800386, Charlottesville, VA 22908, United States; Houpt, E.R., Division of Infectious Diseases and International Health, University of Virginia, PO Box 801340, Charlottesville, VA 22908, United States
Detection of diarrheagenic Escherichia coli (DEC) typically depends on identification of virulence genes from stool cultures, not on stool itself. We developed a multiplex polymerase chain reaction (PCR) assay that detects key DEC virulence genes (stx1, stx2, eae, bfpA, ipaH, LT, STh, aaiC, aatA). The assay involved a multiplex PCR reaction followed by detection of amplicon(s) using Luminex beads. The assay was evaluated on over 100 colony and broth specimens. We then evaluated the assay using DNA extracted from stool, colony pools, and Gram-negative broths, using stool spiked with known quantities of DEC. Performance of the assay on stool DNA was most quantitative, while stool broth DNA offered the lowest limit of detection. The assay was prospectively evaluated on clinical specimens in Tanzania. Stool DNA yielded higher sensitivity than colony pools compared with broth DNA as the standard. We propose using this assay to screen for DEC directly in stool or stool broths. © 2012 Elsevier Inc.