1. PhD LABORATORY POSTING SEMINAR (MLS 818)BY
OLADEINDE BANKOLE HENRY
REG NO: F
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

2. 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

3. 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.

4. 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.

6. 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

7. 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.

8. 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

9. 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

10. SUSCEPTIBILITY PROFILE REPORT OF TEST BACTERUIM

11. NLSI BASED REPORTING OF BACTERIAL SUSCEPTIBILITY

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

13. 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.