Abstract Background InTray® Mueller Hinton Chocolate (MHC) and Colorex™ Screen (CS) plates are in vitro diagnostic devices combining prepared bacteriological agar-media with Biomed’s ‘InTray®’ format (a double sealed-packaging cassette system that provides extended shelf-life 12 months past the date of manufacture). The devices are intended for use with clinical specimens to culture, isolate and identify potentially pathogenic bacteria. This study evaluated MHC and CS device performance at 7 and 13 months past the date of manufacture after product was experimentally challenged with stress replicating a low-resource tropical field clinic scenario. Methods MHC and CS plates were exposed to high temperature (40.5°C ± 0.5) and high humidity (74.5 ± 2.5% relative humidity) for an extended time-period (18 days). After the 18 days of heat treatment, the stress-group plates were stored in the lab at working room temperature (20-25°C) for the remainder of the study. Quantitative differences between control-groups (stored under refrigeration: 2-8°C) and heat-stress treatment groups for both MHC and CS were examined via CFU/mL measurements comparing three different lab-strain bacteria chosen based on clinical sub-culturing prevalence from blood bottles; Escherichia coli (ATCC 25922), Streptococcus pneumoniae (ATCC 6305), and Staphylococcus aureus (ATCC 25923). Briefly, 24-hour (18-30 h) old bacterial colony suspensions (grown on MHC) were prepared in 0.85% saline solution, adjusted to a 0.5 McFarland standard (here defined as 0.08-0.96 Absorbance @ 625 nm), measured in triplicate and diluted. A 20 μL volume from the lowest dilution (2.5 x 103 CFU/mL) was then spread-plated to each plate in order to count colony-forming units (CFU) in the acceptable range of 25-250. Results Quantitative CFU/mL differences between control-groups (stored under refrigeration: 2-8°C) and heat-stress-treatment groups (>40°C and high humidity) for MHC and CS for three different test bacteria (listed above) were not statistically significant when tested at 13 months. The CS stress and control group comparison for S. pneumoniae with a p value < 0.05 at 7 months had a p value > 0.05 at 13 months, suggesting that the difference between the CFU counts may not be significant in biological terms. Conclusions In conclusion, control and heat-stress treatment groups for both MHC and CS met acceptable performance criteria as expected for the products, 13 months after the date of manufacture.