Table 1.
Comparison of main parameters of 4 real-time PCR kits tested
| Manufacturer or source | Name of kit | Target pathogen(s) | Target gene(s) | Kit includes: |
Reagent storage temp (°C) | Final reaction volb (μl) | Vol (μl) of DNA from sample added | No. of cycles used for comparison (no. of cycles recommended by manufacturer) | No. of tests with kit | Apparatus for which kit is available | |||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Ready-to-use mix containing primers and probe | ICa detection | Extraction controls | Positive control | ||||||||||
| In-house | B. pertussis | IS481 | No | No | No | −20 | 20 | 5 | 45 | ||||
| Argene | Bordetella R-gene | B. pertussis | IS481 | Yes | Yes (10 μl IC to add in sample before extraction) | Yes | Yes | −20 | 25 | 10 | 45 (45) | 60 | LightCycler (LC480/LC2.0), SmartCycler, ABI Prism, Rotor-Gene |
| Shanghai ZJ Bio-Tech (distributed in France by BioAdvance) | Bordetella pertussis real-time PCR kit | B. pertussis | IS481 | No | Yes (1 μl IC to add in mix before amplification) | No | Yes (107 copies/ml) | −20 | 25 | 2.5 | 45 (40) | 25 | LightCycler (LC480), iCycler (iQ4/iQ5), SmartCycler II, ABI 7000/7300/ 7500/7900, Rotor-Gene 6000, Mx3000P/ 3005P, MJ Option2/ Chromo4 |
| Cepheid | SmartCycler Bordetella pertussis/parapertussis assay | B. pertussis and B. parapertussis | IS481 (B. pertussis), IS1001 (B. parapertussis) | Yes | Yes (contained in mix [20 copies/μl]) | No | No | +4 (beads) | 25 | 5 | 45 (40) | 40 | SmartCycler II |
| Focus Diagnostics (distributed in France by Eurobio) | Simplexa Bordetella pertussis/parapertussis assay | B. pertussis and B. parapertussis | IS481 (B. pertussis), IS1001 (B. parapertussis) | No | Yes (5 μl IC to add in sample before extraction) | Yes | Yes | −20 | 25 | 5 | 45 (45) | 48 | LightCycler LC480, SmartCycler II, ABI 7500 |
a
IC, internal control.
b
As recommended by the manufacturer of the SmartCycler apparatus.