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The ubiquitous and opportunistic pathogen  Cronobacter sakazakii  is responsible for severe meningitis, sepsis, and necrotizing enterocolitis in neonates and infants associated with ingestion of contaminated powdered infant formula (PIF). The current ISO method for isolation and detection of  Cronobacter  spp. is laborious, time-consuming and expensive. In this study, a multiplexed lateral flow test strip was developed to rapidly detect and simultaneously serotype O1 and O2  C. sakazakii  serotypes. The assay is based on two monoclonal antibodies (MAb) that specifically bind to the lipopolysaccharides (LPS) of these pathogens. The test strip provides results very quickly;  C. sakazakii  could be detected in pure culture within 15 min with a sensitivity of 10 7  CFU/ml. After non-selective enrichment for 18 h as low as one  Cronobacter  cell per g PIF could be detected. Moreover, the established lateral flow assay (LFA) offers excellent specificity showing no cross-reactivity with other  C. sakazakii  serotypes,  Cronobacter  species or  Enterobacteriaceae  tested. These characteristics, together with several advantages such as speed, simplicity in performance, low analysis cost, and no requirement of specialized skills or sophisticated equipment make the developed multiplexed LFA suitable for reliable detection and serotyping of  C. sakazakii  serotypes O1 and O2.

Multiplexed Lateral Flow Test for Detection and Differentiation of Cronobacter sakazakii Serotypes O1 and O2