TABLE 5.
Methods used by participating laboratories with EQA panel 2
| Characteristic | Lab 1 | Lab 2 | Lab 3 | Lab 4 | Lab 7 | Lab 8 | Lab 9 | Lab 10 | Lab 11 |
|---|---|---|---|---|---|---|---|---|---|
| In-house method | Yes | Yes | Yes | Yes | Yes | Yes | Yes | Yes | Yes |
| Concentration of sample | No | No | No | No | No | No | No | No | Centrifugation |
| Extraction method | Boil | Silicaa | GuSCNb | MagnaPurec | Qiagend | Boil | Qiagen | Amplicore | Qiagen |
| Eq. sample used for PCRf | 1 | 5 | 4 | 20 | 8 or 20 | 10 | 12.5 | 5 | 12.5 |
| Amplification method | Single PCR | Real time | Nested PCR | Real time | Real time | Nested PCR | Real time | Real time | Real time |
| Target gene | IS481-IS1001 | ?g | IS481-IS1001 | IS481-IS1001 | IS481-IS1001 | ? | IS481 + ptxA Prh | IS481 | Pertactin |
| Detection method | Gel | Taqmani | Gel | Taqman | Taqman | Gel | FRET probe | Taqman | Taqman |
| Analytical sensitivity (per ml) | 1,000 CFU | Unknown | Unknown | Unknown | 125 CFU | 10 cells | 200 CFU | Unknown | Unknown |
| No. of separate areas | 3 | 3 | 4 | 1 | 3 | 4 | 3 | 3 | 2 |
| Carryover prevention | No | dUTPj | No | dUTP | No | No | dUTP | dUTP | dUTP |
a
Silica extraction method.
b
GuSCN, guanidine thiocyanate lysis and isopropanol precipitation.
c
MagnaPure (Roche Diagnostics).
d
QIAamp DNA mini kit (Qiagen).
e
Amplicor respiratory specimen preparation kit.
f
Equivalent (Eq.) sample used for PCR (in microliters), calculated as the starting volume used for concentration and extraction × fraction of extracted volume added to the amplification mixture.
g
?, participant did not answer.
h
ptxA Pr, pertussis toxin promoter.
i
Taqman double dye probe.
j
Addition of dUTP and uracil glycosylase.