PRIMARY AMOEBIC MENINGOENCEPHALITIS CAUSED BY NAEGLERIA FOWLERI, KARACHI, PAKISTAN

PRESENTED BY :DARAKHSHAN SALEEMMS MICROBIOLOGYFUUASTE

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.

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

SYMPTOMS :

Headache

fever

stiff neck

loss of appetite

vomiting

altered mental state

seizures

comaThere may also be hallucinations, drooping eyelid, blurred vision, and loss of the sense of taste.

Two cases were studied at AKUH

In June 2008

•2 DAYS HISTORY

•HGF

•SEIZURES

•COMATOSE WITH FIXED AND DIALATED LEFT EYE PUPIL

IN SEPTEMBER 2008PATEINT WAS 25 YEARS OLD

• 24 HRS HISTORY

• FEVER

• VOMITING

• NECK RIGIDITY

CSF ANALYSIS IN BOTH CASES

• ELEVATED PROTEIN LEVEL

• ELEVATED NEUTROPHILIC PLEOCYTOSIS

• HYPOGLYCORRHACHIA

• GS AND LA WERE NEGATIVE

TILL APRIL2009 NO PAM DETECTED

IN APRIL 2009 THERE WERE 11

PATIENTS WITH PAM REFERRED TO

AKU

THOSE PATEINTS WERE TREATED WITH

AMPHOTERECIN B

RIFAMPIN,FLUCONAZOLNZOOLRIFA

MOITHEY WERE INCUBATED AND VANTILATED

AFTRE HOSPITAL ADMISSION

BUT NO ONE SURVIVED

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

ON THE BASIS OF THESE DIAGNOSIS

WET MOUNT DONE BY LAB

MOLECULAR DIAGNOSIS:

IN JUNE 2009 CSF COLLECTED FROM 3

PATIENTS AND CONFIRMED THE INFECTION

BY REAL TIME PCR

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

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

• THIS WAS DETECTED BY

HEXACHLOROFLUORESCEIN(HEX)

LABELED PROBE Nfowl(5’-HEX-AT AGC

AAT ATA TTC AGG GGA GCT GGG C-

BHQ1-3’)

PCR PERFORMANCE

2 INITIAL INCUBATIONS

AT 50 C WITH URACIL-DNA-

GLYCOSYLASE ACTIVITY FOR 2

MINS

2ND AT 95 C WITH ACTIVATION

OF PLATINIUM Taq DNA

POLYMERASE)

FLOURESECNCE WAS MEASURED

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