TABLE 1.
Center | Vol (ml) used | RCL | WCL | Fungal lysis | Method of fungal DNA purification | DNA strategy | Internal control PCR | No. of replicates | Compliant | PCR strategy |
---|---|---|---|---|---|---|---|---|---|---|
2 | 3 | Yes | Yes | B. beating | QIAamp Blood kit (Qiagen) | DNA 1 | No | 3 | No | PCR 1 |
3 | 3 | Yes | No | B. beating | MagNA Pure LC Total NA kit (Roche) | DNA 2 | Yes | 3 | Yes | PCR 2 |
4 | 1-2 | Yes | No | Lyticase | Phenol-chloroform | DNA 3 | Yes | 1 | No | PCR 3 |
5 | 3 | Yes | Yes | Lyticase | DNAeasy Tissue (Qiagen) | DNA 4 | No | 3 | No | PCR 4 |
6 | 3 | Yes | No | B. beating | MagNA Pure LC DNA kit I (Roche) | DNA 5 | No | 3 | No | PCR 5 |
7 | 3 | Yes | No | B. beating | ZR Fungal/Bacterial DNA kit (Zymo research) | DNA 6 | Yes | 3 | Yes | PCR 6 |
8 | 1.5 | Yes | No | B. beating | SeptiFast kit (Roche) | DNA 7 | Yes | 3 | No | PCR 7 |
9 | 3 | Yes | Yes | B. beating | SeptiFast kit (Roche) | DNA 8 | Yes | 3 | Yes | PCR 7 |
10 | 3 | Yes | Yes | B. beating | MagNA Pure LC Total NA kit (Roche) | DNA 9 | Yes | 3 | Yes | PCR 1 |
11 | 3 | Yes | Yes | B. beating | Tissue kit, EZ1 (Qiagen) | DNA 10 | Yes | 3 | Yes | PCR 8 |
13 | 3 | Yes | Yes | B. beating | High Pure Template PCR kit (Roche) | DNA 11 | Yes | 3 | Yes | PCR 1 |
14 | 3 | Yes | Yes | B. beating | SeptiFast kit (Roche) | DNA 8 | Yes | 3 | Yes | PCR 9 |
15 | 3 | No | No | B. beating | QIAamp blood kit (Qiagen) | DNA 12 | Yes | 3 | Yes | PCR 1 |
16b | 3 | ND | ND | B. beating | QIAamp DNA minikit (Qiagen) | DNA 13 | Yes | 3 | Yes | PCR 10 |
17 | 3 | Yes | Yes | Lyticase | DNA Lego kit (Top Bio) | DNA 14 | No | 3 | No | PCR 11 |
18 | 3 | Yes | No | B. beating | MagNA Pure LC DNA kit I (Roche) | DNA 5 | No | 3 | No | PCR 5 |
19 | 3 | Yes | Yes | B. beating | GeneXpert (Cepheid) | DNA 15 | Yes | 3 | Yes | PCR 12 |
20 | 3 | Yes | Yes | B. beating | High Pure Template PCR kit (Roche) | DNA 11 | Yes | 1 | No | PCR 2 |
21 | 2 | Yes | No | B. beating | SeptiFast kit (Roche) | DNA 7 | Yes | 3 | No | PCR 1 |
22 | 3 | Yes | Yes | B. beating | High Pure Template PCR kit (Roche) | DNA 11 | Yes | 3 | Yes | PCR 13 |
23 | 3 | Yes | Yes | B. beating | bioMérieux MiniMagc | DNA 16 | Yes | 3 | Yes | PCR 1 |
The results for the centers that used methods that attained the detection threshold are indicated in boldface. These included 11 compliant centers and 3 noncompliant centers. Eleven different DNA extraction techniques were used together with nine different PCR protocols by the 14 centers attaining the threshold, 9 of which were compliant; 8 attained the PCR performance threshold, and 5 achieved enhanced PCR performance. No results were received from centers 1 and 12, whereas logistical limitations prohibited the participation of center 24. Abbreviations: RCL, red cell lysis; WCL, white cell lysis; B. beating, bead beating; ND, not disclosed.
The specificity of the assay was poor (11.1%).
For the extraction reagents, any possible contaminating DNA sources were removed by magnetic silica treatment.