Volume 17, Number 4—April 2011
Research
Molecular Epidemiology of Coxiella burnetii from Ruminants in Q Fever Outbreak, the Netherlands
Hendrik I.J. Roest
, Robin C. Ruuls, Jeroen J.H.C. Tilburg, Marrigje H. Nabuurs-Franssen, Corné H.W. Klaassen, Piet Vellema, René van den Brom, Daan Dercksen, Willem Wouda, Marcel A.H. Spierenburg, Arco N. van der Spek, Rob Buijs, Albert G. de Boer, Peter Th.J. Willemsen, and Fred G. van Zijderveld
, Robin C. Ruuls, Jeroen J.H.C. Tilburg, Marrigje H. Nabuurs-Franssen, Corné H.W. Klaassen, Piet Vellema, René van den Brom, Daan Dercksen, Willem Wouda, Marcel A.H. Spierenburg, Arco N. van der Spek, Rob Buijs, Albert G. de Boer, Peter Th.J. Willemsen, and Fred G. van ZijderveldTable 2
Primers and probes used in the PCR for detecting Coxiella burnetii in clinical samples and loci and primers for MLVA of C. burnetii, the Netherlands, 2007–2010*
| Identification | Temp, °C | Primer sequence for MLVA, with label indicated, 5′ → 3′ |
|
|---|---|---|---|
| Forward | Reverse | ||
| Primers IS1111a | 60 | CATCACATTGCCGCGTTTAC | GGTTGGTCCCTCGACAACAT |
| Probe IS1111a | 60 | AATCCCCAACAACACCTCCTTATTCCCAC | |
| Probe inhibition control | 60 | ACATAATCTCTCCGACCCCACACTTCCATAC | |
| Cbu0448_ms03_12bp_7U_229bp | 60 | 6-FAM-TTGTCGATAAATCGGGAAACTT | CACTGGGAAAAGGAGAAAAAGA |
| Cbu1963_ms21_12bp_6U_210bp | 60 | NED-AGCATCTGCCTTCTCAAGTTTC | TGGGAGGTAGAAGAAAAGATGG |
| Cbu1980_ms22_11bp_6U_246bp | 60 | PET-GGGGTTTGAACATAGCAATACC | CAATATCTCTTTCTCCCGCATT |
| Cbu0259_ms24_7bp_27U_344bp | 65 | VIC-ATGAAGAAAGGATGGAGGGACT | GATAGCCTGGACAGAGGACAGT |
| Cbu0838_ms27_6bp_4U_320bp† | 65 | 6-FAM-GGGTCAGGTGGCGGGTGTG | TTCTCGCAAACGTCGCACTAACTC |
| Cbu0839_ms28_6bp_6U_480bp† | 60 | VIC-TAGAAACCGATAATCCCCTTGACA | ATTCCGCCGCCATTGAG |
| Cbu1351_ms30_18bp_6U_306bp‡ | 60 | NED-ATTTCCTCGACATCAACGTCTT | AGTCGATTTGGAAACGGATAAA |
| Cbu1418_ms31_7bp_5U_285bp‡ | 60 | PET-GGGCATCTAATCGAGATAATGG | TTTGAGAAAATTTTGGGTGCTT |
| Cbu1471_ms34_6bp_5U_210bp | 60 | 6-FAM-TGACTATCAGCGACTCGAAGAA | TCGTGCGTTAGTGTGCTTATCT |
| Cbu1941_ms36_9bp_4U_477bp‡ | 65 | VIC-GAAACCAGTCTTCCCTCAACAG | ATAACCGTCATCGTCACCTTCT |
*MLVA, multilocus variable-number tandem-repeat analyses; temp, annealing temperature.
†Different primer set from the proposed set by Arricau-Bouvery et al. (11).
‡Updated after personal communication with Le Flèche, Université Paris-Sud, Orsay Cedex, France.
