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

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

  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
  1. 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.