Abstract
Biobanking is the collection of biological materials and their related data into an organized system for future scientific use. 1 In Uganda, most biobanks in health centers are managed by research groups that maintain freezers used to store samples such as blood, serum, urine, feces, tissues, DNA, and live infectious agents. The needs of these small biobanks are unknown, thus affecting planning and resource allocation toward sustenance and upgrading them to larger supportive facilities.
In many settings, there have been complaints about failure to recover bacteria, which hinders prospective projects. To follow up on these recurring issues, we analyzed the situation at an anonymous laboratory-based biobank at Makerere University, Uganda. We first confirmed bacterial loss allegations after experimenting to recover a 7-year (2012–2018) collection of 188 bacteria (Escherichia coli; 94 and Salmonella spp.; 94). The bacteria had been isolated from animal clinical samples, and after testing, the isolates were stored in 10% skimmed milk or brain heart infusion (BHI) broth supplemented with 20% glycerol before archiving in a −30°C freezer.
In brief, we thawed the frozen stocks at room temperature and then, 100 μL of the bacterial suspension was pre-enriched in 2 mL of BHI broth (Oxoid, USA) at 37°C for 48 hours. The overnight culture was inoculated on MacConkey agar (Conda laboratories, Spain) and the grown colonies were identified as E. coli or Salmonellae, using methods outlined by Kakooza et al. 2 Overall, 58.5% (110/188) of the strains were successfully regrown (had at least one colony grown after inoculating the primary broth culture).
These outcomes stimulated us to further perform an overview of the laboratory quality management system (LQMS). An internal audit was performed using a score-based questionnaire, harmonized according to the International Organization for Standardization (ISO) 17025 and ISO 15189 standards.3,4 The assessment focused on the essentials, whose degree of implementation could affect the reliability of the biobank. Compliance to the different aspects of the ISO elements was scored on a scale of 0 to 3 (0, no compliance; 1, low compliance; 2, moderate compliance; and 3, high compliance). The total points earned were computed and equated to a percentage. Numerous gaps were exposed after comparing actual performance with the ISO standards.
The gaps in standards compliance included lack of full knowledge, and experience concerning the laboratory standards and their implementation. The absence of established policies and technical procedures was noted. A majority of the key quality documents (quality manual, biosafety manual, standard operating procedures, forms, logs, and others) were still absent. The implementation scores of the essentials were organization and management needs (5/24; 20.8%), facilities and safety (21/48; 43.8%), equipment (14/60; 23.3%), purchasing and inventory (10/36; 27.8%), process management (38/81; 46.9%), assessments (3/45; 6.7%), personnel (13/75; 17.3%), customer focus (2/15; 13.3%), nonconforming incident management (2/21; 9.5%), continual improvement (3/33; 9.1%), document control (5/48; 10.8%), information management (5/24; 10.8%), and overall compliance (121/510; 23.7%).
It is important to address needs and the status of laboratory-based biobanks for proper management, which we attempted to do. We were limited to pilot experiments. We hypothesize that (1) bacterial species possess variable survival capabilities, (2) the type of preservation media used may also affect cryopreservation, and (3) the high bacterial loss rate could also be due to the minimal establishment of sufficient LQMS in this setting. For better outcomes, we recommend modifications in the bacterial preservation and revival methodologies done at the laboratory. The views expressed are those of the authors, thus a country-wide survey on the status of biobanks and their needs would give a better overall picture, since this was a small study.