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

BMC Microbiology

Table 1 Balamuthia mandrillaris RNase P genomic DNA detected by real-time PCR – specificity and detection limits

	DNA [pg] ^a)	Amoebae equivalents ^b)	CT ^c)
B. mandrillaris (axenic culture)	5 – 50	< 2	35 – 38
Acanthamoeba hatchetti 2HH (axenic culture)	≥ 500	neg	≥ 39
A. lenticulata 72/2 (axenic culture)	≥ 500	neg	≥ 39
A. castellanii 1BU (axenic culture)	≥ 500	neg	≥ 39
Toxoplasma gondii (partially purified)	≥ 500	neg	≥ 39
Leishmania major (axenic culture)	≥ 500	neg	≥ 39
Pneumocystis murina (partially purified)	≥ 500	neg	≥ 39
Mycobacterium bovis (BCG)	≥ 500	neg	≥ 39
Murine macrophage RAW 264.7 cells (TIB 71)	≥ 500	neg	≥ 39
Human neuroblastoma Kelly cells (ACC 355)	≥ 500	neg	≥ 39
Brain tissue (mouse, ca. 15 mm³)	≥ 500	neg	≥ 39
Brain tissue spiked with 1–10 B.m.	n.d.	1 – 10	33 – 38
Lung, liver, spinal chord (mouse, ca. 15 mm³)	> 500	neg	≥ 39
Lung tissue spiked with 10 B.m.	n.d.	10	35–36

^a) Minimal amounts of total DNA giving positive results (CT < 39). ^b) 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. ^c) Range of CT values measured; neg = CT ≥ 39 or no DNA detected; n.d. = not done.

ISSN: 1471-2180