Uriselecttm 4

When not stated otherwise, chemicals, reagents and laboratory utensils were procured from Thermo Fisher Scientific. Bacteria were cultured onto blood (Merck KGaA) or UriSelect^TM 4 (Bio-Rad) agar, and CAMHB (Becton, Dickinson and Company) was used in all suspension cultures except for BMD. Sterile filtration was performed using Nalgene filter units (PES 0.1 μm, VWR International LLC).

The required antibodies were produced on a large scale as previously described¹⁹ (link) and provided to NG Biotech (Guipry, France) for the development of multiplex LFIA (see Figure 1). The strains to be tested were grown overnight at 37°C on URISelect^TM 4 or Mueller–Hinton agar plates (Bio-Rad, Marnes la Coquette, France). One single isolated colony was collected from the plate with an inoculation loop and suspended in 150 μL of extraction buffer to perform the lysis step. Subsequently, 100 μL of this extract was loaded on the cassette and allowed to migrate for 15 min. The protocol of the commercialized kit will be similar to the protocol described here.