TABLE 2.
Effectiveness of V. vulnificus detection in gulf water supplemented with 0.2% (wt/vol) peptone by real-time PCR with various DNA extraction methods
| DNA extraction method | Protocol summary | Sensitivity of detection (gulf water) (CFU/ml)a |
|---|---|---|
| Boiling | Boil at 100°C for 10 min | ≥103 |
| Boiling with Instagene (Bio-Rad) | Incubate at 56°C for 20 min, followed by boiling for 10 min | ≥102 |
| Qiaprep (Qiagen, Valencia, Calif.) | Bacterial pellet resuspended in buffer P1; buffers P2 and N3 added; ethanol precipitation | ≥107 |
| Magnetic Beads (Bugs n' Beads, Genpoint, Oslo, Norway) | Incubate with binding beads; lysis buffer added; ethanol precipitation | ND |
| Nucleotrap DNA purification (Clontech, Palo Alto, Calif.) | Buffer NT2 and Nucleotrap suspension added to DNA extracted by boiling; buffer NT3 added to precipitate DNA; DNA eluted by incubation at 50°C for 5 min | ND |
a
Results of three consecutive experiments which produced consistent results. ND, no detection.