Makerere University College of Health Sciences has a state of the art research laboratory on the first floor of the Clinical Research Building outfitted with modern equipment. Its clinical capabilities include testing of HIV, Hepatitis B, Herpes Simplex 2, and syphilis, Pregnancy etc. Research capabilities include PBMC isolation and cryopreservation, cell culture, interferon gamma release assays (IGRA), Whole Blood and Interleukin Assays involving cell processing for intra cellular cytokine assays, T-Spot among others. The laboratory policy is to maintain at least four IATA certified staff to handle specimen shipments. It also provides quality laboratory services to several external projects and research clinics particularly sample processing, storage and shipment.
Staff - The Laboratory team has nine well-trained staff (Director, six technologists, one Manager and a Data Manager) led by the Laboratory director, Assoc. Prof Moses Joloba. The laboratory policy is to maintain at least four IATA certified staff to handle specimen shipments. In addition, the Immunology Laboratory has been an occupational trainer to many students in a large range of immunological research areas. Steve Cose PhD, a resident Immunology consultant, is available for consultation and technical supervision of immunology research techniques ongoing in the Laboratory. All staff are Human Subject Protection trained(HSP) and Good Clinical Laboratory Practice (GCLP) trained.
Equipment - The Immunology Lab is well equipped with up-to-date and current testing requirements. All lab equipment are modern, our testing methodologies are based on up-to date medical technology and all Lab equipment are well serviced and covered by a service contract. Currently there are primary and back up equipment for micro-centrifuges (Haraeus 17), routine centrifuges (Labofuge 400R & sigma), EIA washer (SEAC radim&Dynex MRW), ELISA readers (Biomerieux), precision pipettes (Gilson &Finnpipett), incubators(memmert), CO2 incubator (Innova&Sannyo), refrigerators (4 types), Freezers (--20C Danfrost& -80C New Brunswich), biohazard flow hoods(sterilGARD), fume extract hood (Structural inbuilt), programmable water baths (memmert), heat blocks (Bioer), incubator shakers (Bioer), spectrophotometers (Jenway), vortex machines(Vortex Genie), microscopes (Olympus), shakers (Cyan), Cryoshippers(MVE) and Liquid LN2 torageDewars (MVE & Taylor Wharton), and a FACS canto II flow cytometer (BD).
Quality management (QC/QA/QI) - The quality assurance program covers the pre-analytical, analytical, and post analytical phases. This is through standard procedures like sample rejection/acceptance, routinely monitored Turn Around Time, Internal Quality Controls (IQCs), EQA proficiency programs, Blind/Split sample testing etc. The Laboratory is currently in its advanced stage preparations for accreditation for SANAS/SADCAS accreditation. Other Key Players aiding Quality at the Immunology Laboratory Contact Laboratory Services (CLS), South Africa monitors the laboratory three times to ensure International standards are adhered to.
The Molecular Biology laboratory
This laboratory was established with support from NIH and SIDA (Sweden). It is approx. 800 sq. ft., air conditioned well equipped and appropriately organized for most molecular biology oriented studies. The laboratory has acquired an additional 2500 sq. ft. space (with offices and a seminar room) that will house the human genetics laboratory. Equipment and software: The laboratory has a real time thermocycler (Rotor Gene Q 6000), two incubator shakers (Innova 3100), incubator (TwinCubator) , hybridization oven (Labnet), four refrigerators, microcentrifuge (Eppendorf), belly dancer (Stovall), electroporator (BioRad), conventional thermocyler (Techne TC-412), three gel electrophoresis systems (), two UV transilluminators, a polaroid camera (UVP), gel imaging system (BioDoc-It), a blotting apparatus, incubator (Binder), pH meter (Oakton), analytical balance (Sartorius), autoclave (Priorclave), an ice maker, sterilizing hood (Cleanspot), crosslinker (Spectrolinker), spectrophotometers (GeneQuant, Cecil, Janeway 3605), five internet linked computers, and a dark room. Software are installed and operational including the Bionumerics for molecular analyses.
This laboratory performs most basic, applied and molecular epidemiology techniques. Freezing space - currently we have 2 -80C freezers chest and upright with capacity to hold 50,400 and 30,000 samples (in 1.5 - 2mL cryovials) respectively. We have 2 -20C freezers chest and upright with capacity to hold 23,300 and 22,600 samples (in 1.5 - 2mL cryovials) respectively. There are 50,000 samples stored in the freezer space. These are mainly samples generated from our studies or referred for further testing. The molecular biology laboratory stores about 5000 samples a year. Data handling- a laboratory information system is used to keep records on the archival, storage and retrieval of the stored samples. All equipment are under a renewable annual service contract and are serviced according to schedule. Power back up- The laboratory is power backed up by two interconnecteted automatic generators. The -80C freezers, the main DNA biorepositories are in addition backed up by 4hour UPS and a 24 hour carbondioxide tank in case of mechanical failure.
The Molecular Diagnostic Laboratory
The space is located on the top floor of the Pathology/ Microbiology building that includes 225 sq/ft for the pre- amplification room, 240 sq/ft for the amplification laboratory, 340 sq/ft for the post-amplification room, 120 sq/ft for office space and 200 sq/ft for the boardroom with video conferencing facilities installed and functional. Equipment available in this facility include Rotor Gene real-time PCR cycler, spectrophotometer (NanoDrop 2000c), DNA extraction machine (QIAcube), QIAxcel robotic PCR setup (liquid handling), Pipetteman, electrophoretic equipment (BIORAD), hybridization oven (ProBot), -80C freezers (Brunswick), fridges and automated electrophoretic system (QIAgility). Freezing space - currently we have 2 -80C freezers chest and upright with capacity to hold 50,400 and 30,000 samples (in 1.5 - 2mL cryovials) respectively. We have 2 -20C freezers chest and upright with capacity to hold 23,300 and 22,600 samples (in 1.5 - 2mL cryovials) respectively. Approximately one third of this total freezer space is available for the feasibility study. Offices are equipped with computers installed with up-to-date software used for analysis.
The facility is air conditioned maintaining ambient temperature and keeping it dust free. Storage capacity - for the -80C storage we have the one INNOVA Eppendorff model U725G. The temperature range is -65C to -86C. The the exterior dimensions are 195 x 102.5 x 86.7 cm. The gross weight is 799Ibs. The capacity is 725L which can hold 5042 boxes, 3603 boxes and 2164 boxes. With a 2.0mL sample capacity of 100 samples per box with full capacity of 50,000 samples. All these laboratories are connected to a 100 kVA backup generator (Broadcrown), which is backed up by a 27 kVA generator (Olympian). This ensures continuous availability of electricity during power blackouts. MakCHS has established a nitrogen plant that ensures access to nitrogen required for cryopreservation and shipment of certain specimen types.
Mycobacteriology (BSL-III) Laboratory
The PI, Prof. Joloba directs this Biosafety Level III (BSL-III) TB culture facility operating since June 2009; with controlled uni-directional flow of HEPA filtered air. It has a level II section and a containment (Level III) section, two office rooms, a reception area, a well-monitored store and a training room. The facility is approximately 90 square meters overall in size and is well monitored with a 24-hour, 7-day per week surveillance system (intelligent Video Management system) for the BSL-II and BSL-III sections. It has separate, well-equipped and operational Immunology and Molecular Biology sections. The BSL-III facility has a dedicated facility manager with 8 years TB-related experience as well as 10 additional BSc level technologists, an MSc level data manager, a quality assurance officer, safety officer, laboratory training coordinator and additional facilities assistants. All personnel are up-to-date with advances in mycobacteriology/molecular and immunology assays as well as quality systems management aspects.
Personnel participate in national and international EQA panels (i.e., CAP EQA- pSMILE, NHL/NCID, New York state Department of Health) as well as inter-laboratory proficiency work with the National TB Reference Laboratory and FIND Laboratory in Kampala. This laboratory is the site of the Aeras and NIH TB vaccine trials in Uganda. The facility is housed within the College of Health Sciences on the Makerere University campus, adjacent to Mulago Hospital campus TB and other inpatient/outpatient clinics. The facility is 1 kilometer from the Uganda National TB Reference Laboratory- Wandegeya and 8 kilometers from the Joint Clinical Research Centre (JCRC) clinics and TB Laboratory on the Mengo campus and 20 kilometers from the JCRC clinics and laboratories at the Lubowa campus. The Mycobacteriology Laboratory has back- up agreements in place with both the National TB Reference Laboratory and JCRC TB laboratory (separately described below).
Equipment - This facility has two MGIT 960 machines, one Bactec 9120, three 9050 blood culture systems, four 6-foot class II & one 4-foot class II biosafety cabinets, four autoclaves, four hot air ovens, four -800c carbondioxide freezers, six -200c freezers, ten refrigerators, four CO2 incubators, two Hain twin incubators, one automated DNA extraction machine, two four module GeneXpert machines, four refrigerated centrifuges-Allegra 12R, four weighing balances, two PH meters, one Tachometer, one BX51 research fluoresce microscope, one LED microscope, six light microscopes, four water baths, one microfuge, two sonicating baths, ten desktop computers, four LCDs and two screens, four laptop computers, a freezer works ( barcode printer and scanner), 2 photocopiers, central UPS and stabilizers, and 2 back-up 100KVA generators. The laboratory data system is well equipped with recent technologies including Freezerworks system for sample management.
Current capacity and activities routinely include light and fluorescent microscopy, sputum and gastric aspirates culture on solid media (LJ, 7H10/11, 7H9) including quantitative and qualitative methods as well as liquid (MGIT 960), and blood cultures using Bactec 9120 and 9050 culture systems. Mycobacterium identification, speciation and epidemiological assays are done using PCR IS6110, Capilia TB NeoTM and Hain Genotype Assay, RFLP, MIRU-VNTR, spoligotyping, and SNP analysis. Drug susceptibility testing (DST) for first line (SIREPZA), LPA, MODS and second line drugs using MGIT and APM are also routinely performed.
Liquid Nitrogen plant
Makerere University College of Health Sciences has a liquid nitrogen plant located on site. It has two StirLIN-1 Compact that produce 240 l/day on site LN2 supply. Has power consumption of 16 kW, connected to a 3 phase power supply at 200 to 480 V, 50/60 Hz, cooling water consumption of 1000l/h at 15C located in adequate space. It has a noise level 70 dBA as is environmentally friendly. Plant room temperature is maintained at 25C, altitude 250m, relative humidity 95%, and cooling water temperature 15C. Has a liquid nitrogen storage capacity of 300 l, with a level indication and control.
It has a dedicated generator set for power supply back up, connected to voltage stabilizer for utility power stabilization. The personnel posses liquid nitrogen handling personal protection materials and clothing. The plant meets European CE safety standard, IP54, IEC 60204 and other international standards. It has an integrated system diagnostics, fully automatic start/stop and restart after power failures, no defrosting or purging required, 10 minutes start-up time to full production, single switch operation.
Proposed IBRH3AU Infrastructure Upgrade
The MakCHS has dedicated up to 6500 sq. ft. of space for alterations and renovations to provide adequate infrastructure of a world-class biorepository in Africa.
Space - up to 6500 sq. ft. of space has been dedicated and for the IBRH3AU.
Specialized rooms - among these are a freezer room, nitrogen freezer room, automated DNA/RNA extraction/PCR-set up laboratory, post-PCR laboratory, live cell preparation and freezing laboratory, blood processing laboratory, office space and computer/database laboratory.
Access - a role-based access control system that restricts access to the facility to authorized personnel only will be installed; we shall install electronic finger print access control each major entry and exit points.
Water purification -we shall install a dedicated RO water purification plant for the IBRH3AU laboratories with capacity of 1000LPH, an ISBER recommendation that repositories maintain separate water purification services.
Heating, ventilation, and air-conditioning - we propose to install a Heating, Ventilation, and Air-Conditioning (HVAC) system to maintain good indoor air quality through adequate ventilation with filtration and provide proper thermal control.
Electrical set-up - the IBRH3AU is connected to a 3 Phase electricity line which is capable of supporting the heavy equipment. The equipment will also be connected to UPS system to protect them from power surges. We already have two stand-by generators that will support the facility during periods of power outage. Plans have been made to add a third backup generator that will be dedicated to the biorepository.
Equipment - this Phase will be earmarked with transition to automation and thus equipment are planned. Five years ago the QIAcube was installed in the Molecular Biology laboratory, the equipment has a robotic workstation for automated extraction and purification of DNA, RNA and protein with capacity for 12 biospecimens per run. With previous use of such equipment we possess the experience in deploying and use of automated technology. Therefore, we shall smoothly transition high volume automated equipment. Proposed equipment include Agilent Bravo liquid handler platform, QIAsymphony, freeze dryer and Agilent 2100 Bioanalyzer. Other equipment considered for the IBRH3AU -80C freezers, -20C freezers, laminar flow hoods, vapor-phase liquid nitrogen freezers, high speed centrifuges, dry ice maker, nitrogen shippers, autoclave and incubators.
Computers - an IT room will be set up and equipped with computers with the latest software and informatics systems for the biorepository. A computing resource will be put in place capable of handling the minimum data set and any analyzed genomic data annotated to biospecimens. This will comprise of a workstation with up to 5 CPUs and 8-16 GB of RAM.
Back-up plans - freezers and vapor-phase liquid nitrogen freezers storage tanks will be under continuous temperature surveillance and in case of failure the alarm system installed will provide information via calls to the Quality Manager who will coordinate an immediate and corrective action response. As first line back up we shall have CO2 tanks for -80C freezers, and the proposed LN2 tanks attached to the LN2 Freezers are designed to automatically maintain pressure at a pre-set level and supply gaseous products via an internal gas vaporizer maintaining ultra-low temperatures providing time for the emergency response team to plan and execute a effective response.
We shall maintain a set of back up freezers in the event that we need to move biospecimens because of freezer break down. Off-site backup is a key requirement for an international repository; the IBRH3AU management is negotiating an agreement with the National Health Laboratory Center to provide an off-site backup repository. We shall also explore using the other H3Africa biorepositories as off-site back up.