PhD LABORATORY POSTING SEMINAR (MLS 818) BY OLADEINDE BANKOLE HENRY REG NO: 2012617004F DEPARTMENT OF MEDICAL LABORATORY SCIENCE (MEDICAL MICROBIOLOGY),FACULTY OF HEALTH SCIENCE AND TECHNOLOGY, COLLEGE OF HEALTH SCIENCES, NNAMDI AZIKIWE UNIVERSITY,NNEWI CAMPUS SUPERVISOR PROF. M.I EKEJINDU

OUTLLINE Introduction Patient information Processing of specimen Protocol for viral RNA extraction Primers used for amplification Reaction mixture constituents Thermal cycling profile and amplicon detection on gel agarose Assessment of blood for Lassa virus Interpretation of result Conclusion and Recommendation

INTRODUCTION Urinary tract infections are among the most common bacterial infections in humans both in the community and hospital settings, and they occur in all age groups, usually required urgent treatment (Oladeinde et al., 2011) Effective management of urinary tract infections depends largely on the availability of timely and accurate Laboratory data. Accurate identification and precise reporting of susceptibility profile of uropathogens is critical in assisting physicians in making the right decisions on treatment of patients.

INTRODUCTION CONTINUED Multidrug resistant bacteria isolates are on the rise (Orrett, 2001). Regular monitoring of bacterial isolates for antibiotic resistance development and accurate reporting of same can assist health managers in making right decisions to halt the trend For reliability of test result, susceptibility testing of bacteria isolates should be done in line with guidelines of the Clinical Laboratory Standard Institute (Wilson and Gaido et al., 2004). Deviation from this standard will result in release of laboratory result with little benefit to patients.

PATIENT INFORMATION A 17 year old female patient at the female ward presenting with severe abdominal pain with painful micturation. Provisional diagnosis - Pyelonephritis Investigation required – Urine M/C/S

LABORATORY ANALYSIS Clean– catch midstream urine was collected from the patient into sterile screw-capped universal container, containing few crystals of boric acid as preservative. The specimen was mixed, labeled and transported to the laboratory for processing A loopful (0.001mL) of well mixed uncentrifuged urine was streaked on to the surface of blood agar and cysteine lactose electrolyte deficient medium (CLED) medium and McConkey Agar. The plates were incubated aerobically at 37 0C for 24 hrs.

PROCESSING OF SPECIMEN CONTINUED Five ml of each well –mixed urine sample was centrifuged at 2000g for 5 minutes. The supernatant was discarded and a drop of the deposit was examined microscopically at high magnification for pus cells, epithelial cells, cast, crystals, yeast-like cells, and trichomonas vaginalis Pus cells ≥ 5 per high power field were considered significant to indicate infection

IDENTIFICATION OF UROPATHOGEN The inoculated media were examined after incubation for the presence of bacteria. Resulting bacteria on media were identified using standard bacteriological methods prescribed by Cowan and Steel (1993). RESULT CULTURE: Yielded growth of Escherichia coli after 24 hours incubation

SUSCEPTIBILITY PROFILE REPORT OF TEST BACTERUIM

NLSI BASED REPORTING OF BACTERIAL SUSCEPTIBILITY

INTERPRETATION OF RESULT The bacteria isolate from patient was sensitive to Levofloxacin (LBC) and Ofloxacin (OFX) only

CONCLUSION AND RECOMMENDATION Reading of susceptibility profile of uro-pathogen from patient by Laboratory personnel without recourse to standard zone inhibition measurement resulted in test bacterium incorrectly reported as being sensitive to Ceftazidine (CRO), Gentimycin (GN) and Nitrofurantoin (F). Administration of these antibiotics may result in treatment failure of urinary tract infection in female patients, as well as development of drug resistant strains of bacteria. Strict adherence of Laboratory personnel to stipulated guidelines for reporting susceptibility profile of bacteria isolates incriminated in infectious diseases is advocated.