Detection of Balamuthia mandrillaris DNA by real-time PCR targeting the RNase P gene

BMC Microbiology

Table 2 B. mandrillaris RNase P genomic DNA in infected brain tissues detected by real-time PCR

	DNA [pg] ^c)	Amoebae equivalents ^d)	CT ^e)
Murine brain tissue from non-infected mice	> 500	neg	≥ 39
Murine brain tissue from B.m.-infected mice ^a)	n.d.	n.d.	30
Human brain tissue from non-infected patients	n.d.	neg	≥ 39
Human brain tissue spiked with 50 B.m.	n.d.	50	29
Human brain tissue from BAE-patient 1 ^b) (1:100)	n.d.	n.d.	30
Human brain tissue from BAE-patient 2 ^b) (1:10)	n.d.	n.d.	35

^a) A highly symptomatic CD4⁺ T cell-deficient mouse 14 days post intranasal infection with B. mandrillaris amoebae. ^b) DNA extracted from 2 fresh BAE-patient brain biopsy samples ([23] Table 1: cases 4 and 5). ^c) Minimal amounts of total DNA giving positive results (CT < 39). ^d) Minimal numbers of amoebae (calculated as amoeba DNA equivalents per PCR tube) giving positive results; neg = CT ≥ 39 for at least 500 ng total DNA. ^e) Range of CT values measured; neg = CT ≥ 39 or no DNA detected; n.d. = not done.

ISSN: 1471-2180