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J Med Microbiol 53 (2004), 1007-1012; DOI: 10.1099/jmm.0.45721-0
© 2004 Society for General Microbiology
ISSN 0022-2615

Post-mortem culture of Balamuthia mandrillaris from the brain and cerebrospinal fluid of a case of granulomatous amoebic meningoencephalitis, using human brain microvascular endothelial cells

Samantha Jayasekera1, James Sissons1, Julie Tucker2, Claire Rogers2, Debbie Nolder2, David Warhurst2, Selwa Alsam1, Jonathan M.L. White3, E. M. Higgins3 and Naveed Ahmed Khan1

1School of Biological and Chemical Sciences, Birkbeck College University of London, London WC1E 7HX, UK 2Diagnostic Parasitology Laboratory, London School of Hygiene and Tropical Medicine, Keppel Street, London, UK 3Department of Dermatology, King's College Hospital, London, UK

Correspondence Naveed Ahmed Khan n.khan{at}sbc.bbk.ac.uk

Received April 28, 2004
Accepted July 2, 2004

The first isolation in the UK of Balamuthia mandrillaris amoebae from a fatal case of granulomatous amoebic meningoencephalitis is reported. Using primary cultures of human brain microvascular endothelial cells (HBMECs), amoebae were isolated from the brain and cerebrospinal fluid (CSF). The cultures showed a cytopathic effect at 20–28 days, but morphologically identifiable B. mandrillaris amoebae were seen in cleared plaques in subcultures at 45 days. The identification of the organism was later confirmed using PCR on Chelex-treated extracts. Serum taken while the patient was still alive reacted strongly with slide antigen prepared from cultures of the post-mortem isolate, and also with those from a baboon B. mandrillaris strain at 1 : 10 000 in indirect immunofluorescence, but with Acanthamoeba castellanii (Neff) at 1 : 160, supporting B. mandrillaris to be the causative agent. If the presence of amoebae in the post-mortem CSF reflects the condition in life, PCR studies on CSF and on biopsies of cutaneous lesions may also be a valuable tool. The role of HBMECs in understanding the interactions of B. mandrillaris with the blood–brain barrier is discussed.

Abbreviations: CSF, cerebrospinal fluid; GAE, granulomatous amoebic meningoencephalitis; HBMECs, human brain microvascular endothelial cells; LDH, lactate dehydrogenase.




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