Rat-Bite Fever: Developing and testing a modified time-kill protocol based on Edberg and Edberg 1983 experiemnt in order to test S. moniliformis growth in 0.05% SPS and proposed growth curve

Abstract Introduction/Objective Streptobacillus moniliformis is the primary causative agent of rat-bite fever (RBF) and Haverhill (HF). Rat-bite are difficult to diagnose in a clinical setting likely severely under-represented under-reported worldwide. Clinical presentation often includes fever, chills, myalgia, headache, vomiting. Patients may also develop maculopapular rash covering their extremities approximately 2-4 days after onset followed by polyarthritis roughly 50% patients with mortality rate 10-13% if left untreated. This further complicated fact that submission processing guidelines samples for recovery organism based on ‘outdated’ techniques laboratory methods over 50 years removed from current procedures, instrumentation guidance. Methods/Case Report DNA collected frozen time-point samples, were (n=84) extracted using an in-house custom protocol utilizing 180 µl bacterial lysis buffer 20 PCR grade proteinase K, total 200 µl. The blood culture then thawed ice, sample was added mix, vortexed incubated at 65°C 10 minutes (an additional 95°C inactivation step omitted avoid whole clotting). After incubation, each briefly again Roche automated MagNA Pure Compact instrument initial input volume 400 stored final elution 100 All quantatitive time kill data colleced via CDC designed assay developed S. moniliformis. Results (if Case Study enter NA) Experimenting varying amounts inoculum, ml determined provide best results detection growth/viability as well propose theoretical growth curve organism. Conclusion We found 100% isolates tested (and all variations testing within), SPS (up concentration 0.05 % w/v) commercially available bottles appeared be inactivated, allowing culturing continuous system when inoculated.