Table 4.
Summary of Methods for Detecting Legionella and Two Important Amoebae in Reclaimed Water.
| Organism | Detection Method a | Aggregate Score | Round-Robin Tested? | Growth Media | Key Refs. |
|---|---|---|---|---|---|
| Acanthamoeba | Membrane filtration and incubation | 29 | No | Non-nutrient agar and peptone yeast extract glucose (PYG) | [48] |
| Membrane filtration, enrichment on Neff's media and PCR | 31 | No | Neff's saline non nutrient agar and peptone yeast extract glucose (PYG) | [49,50,51] | |
| Centrifugation and incubation | 29 | Yes | Non-nutrient agar | [52,53] | |
| Membrane filtration, plaque formation and PCR | 30 | No | Non-nutrient agar | [54] | |
| Naegleria | Membrane filtration (or centrifugation) and incubation | 31 | No | Non-nutrient agar and peptone yeast extract glucose (PYG) | [55] |
| Membrane filtration, enrichment, and PCR | 34 | No | Neff's saline non-nutrient agar with E. coli lawn | [55] | |
| Membrane filtration, plaque formation, and PCR | 32 | No | Non-nutrient agar | [54] | |
| Enzyme-linked immunosorbent assay (ELISA) | 38 | No | Non-nutrient agar | [56] | |
| Isoenzyme electrophoretic focusing (IEF) | 32 | No | Non-nutrient agar | [56,57] | |
| Restriction fragment length polymorphism (RFLP) | 32 | No | Non-nutrient agar | [56,58] | |
| Concentration by centrifugation and culture | 39 | No | Non-nutrient agar | [59,60] | |
| Real-time PCR | 40 | No | Not applicable | [61,62,63] | |
| Concentration by centrifugation and then nested PCR | 30 | No | Nutrient agar | [64,65] | |
| Legionella | MF, heating, acidification and plating on CYE | 37 | No | CYE agar | [66] |
| MF, acidification and plating on BCYE | 37 | No | BCYE agar | [28,67] | |
| Direct fluorescent-antibody (DFA) staining | 36 | No | Not applicable | [67] | |
| PCR with EnviroAmp kit | 47 | No | Not applicable | [67,68] | |
| Semi-nested PCR | 41 | No | Not applicable | [69] |
a Detection method in bold were most comprehensive based on an aggregate scoring exercise (see details in [2]).