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.