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NAEGLERIA
- 1. PRIMARY AMOEBIC MENINGOENCEPHALITIS CAUSED BY NAEGLERIA FOWLERI, KARACHI, PAKISTAN PRESENTED BY :DARAKHSHAN SALEEM MS MICROBIOLOGY FUUASTE
- 2. Brain-Eating Amoeba Brain-Eating Amoeba So-called Brain-eating amoeba is a species discovered in 1965. There are several species of Naegleria but only the fowleri species causes human disease. Naegleria loves very warm water. It can survive in water as hot as 113 degrees Fahrenheit.
- 4. N. fowleri amoebas are relatively common, they only rarely cause brain disease. N. fowleri disease is known as Primary Amoebic Meningoencephalitis (PAM). It occurs from zero to eight times a year, almost always from July to September
- 5. SYMPTOMS : Headache fever stiff neck loss of appetite vomiting altered mental state seizures coma There may also be hallucinations, drooping eyelid, blurred vision, and loss of the sense of taste.
- 6. Two cases were studied at AKUH In June 2008 •2 DAYS HISTORY •HGF •SEIZURES •COMATOSE WITH FIXED AND DIALATED LEFT EYE PUPIL
- 7. IN SEPTEMBER 2008 PATEINT WAS 25 YEARS OLD • 24 HRS HISTORY • FEVER • VOMITING • NECK RIGIDITY
- 8. CSF ANALYSIS IN BOTH CASES • ELEVATED PROTEIN LEVEL • ELEVATED NEUTROPHILIC PLEOCYTOSIS • HYPOGLYCORRHACHIA • GS AND LA WERE NEGATIVE
- 9. TILL APRIL2009 NO PAM DETECTED IN APRIL 2009 THERE WERE 11 PATIENTS WITH PAM REFERRED TO AKU
- 10. THOSE PATEINTS WERE TREATED WITH AMPHOTERECIN B RIFAMPIN,FLUCONAZOLNZOOLRIFA MOITHEY WERE INCUBATED AND VANTILATED AFTRE HOSPITAL ADMISSION BUT NO ONE SURVIVED
- 11. BECAUSE…..GS AND LA WERE NEGATIVE CSF CULTURED ON NON NUTRIENT AGAR ON A LAWN OF E.COLI ,OBSERVED CULTURED AMOEBA..WHICH LATER ON PRODUCED FLAGELA
- 12. ON THE BASIS OF THESE DIAGNOSIS WET MOUNT DONE BY LAB
- 13. MOLECULAR DIAGNOSIS: IN JUNE 2009 CSF COLLECTED FROM 3 PATIENTS AND CONFIRMED THE INFECTION BY REAL TIME PCR
- 14. o Separation of DNA double-stranded template o Primer formation o Extension of new DNA strands by a DNA polymerase and deoxyribonucleoside triphosphates (dNTPs) o Other proteins involved PRINCIPLE
- 15. CONT…. PRIMER NaegF192 (3’-GTG CTG AAA CCT AGC TAT TGT AAC TCA GT-5’) AND NaegIR344 (5’-CAC TAG AAA AAG CAA ACC TGA AAG G-3’) WERE USED TO AMPLIFY A 153-bp FRAGMENT
- 16. • THIS WAS DETECTED BY HEXACHLOROFLUORESCEIN(HEX) LABELED PROBE Nfowl(5’-HEX-AT AGC AAT ATA TTC AGG GGA GCT GGG C- BHQ1-3’)
- 17. PCR PERFORMANCE 2 INITIAL INCUBATIONS AT 50 C WITH URACIL-DNA- GLYCOSYLASE ACTIVITY FOR 2 MINS
- 18. 2ND AT 95 C WITH ACTIVATION OF PLATINIUM Taq DNA POLYMERASE) FLOURESECNCE WAS MEASURED
- 19. CONCLUSIONS • PAM CASES INCREASE IN KARACHI AND MOST PATIENTS WERE YOUNG HEALTHY ADULTS . • PAM DIAGNOSIS SHOULD BE IMPROVE BUT THIS IS NOT THE SOLE REASON • PAM IS ASSOCIATED WITH FRESH WATER SWIMMING POOL HAVING POORLY CHLORINATED WATER
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