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Volume 15, Number 4—April 2009

Letter

Multigenotype Q Fever Outbreak, the Netherlands

Corné H.W. KlaassenComments to Author , Marrigje H. Nabuurs-Franssen, Jeroen J.H.C. Tilburg, Maurice A.W.M. Hamans, and Alphons M. Horrevorts
Author affiliations: Canisius Wilhelmina Hospital, Nijmegen, the Netherlands (C.H.W. Klaassen, M.H. Nabuurs-Franssen, J.J.H.C. Tilburg, A.M. Horrevorts); Radboud University Medical Centre, Nijmegen (M.H. Nabuurs-Franssen); Food and Consumer Product Safety Authority, Eindhoven, the Netherlands (M.A.W.M. Hamans)

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Table

Genotyping results for human and animal clinical samples, Q fever outbreak, the Netherlands

Patient/animal no. Sample Ms27* Ms28* Ms34 * Symptoms Ct value† Location
Patient 1
Plasma
3
3
8
Severe
34.4
1
Ewe 1 Vaginal swab 3 3 8 None 25.7 1
Ewe 2 Vaginal swab 3 3 8 None 16.3 1
Ewe 3
Vaginal swab
3
3
8
None
18.8
1
Lamb 1 Throat swab 3 3 8 None 27.9 1
Lamb 2 Throat swab 3 3 8 None 29.9 1
Lamb 3
Throat swab
3
3
8
None
28.9
1
Patient 2 Urine 3 3 7 None 31.7 2
Throat swab 3 3 7 31.8
Patient 3 Urine NR‡ 3 4 Mild 36.7 2
Patient 4 Sputum 4 3 7 Severe 34.2 2
Patient 5
Sputum
3
3
7
Severe
31.9
3
Nine Mile Reference strain 4* 6* 5*

*The allele-calling convention used was as published (3), resulting in a 4, 6, 5 code assigned respectively to the 6-bp repeat unit loci Ms-27, Ms-28, and Ms-34 for the genome sequence of the Nine Mile RSA-493 strain (GenBank accession no. NC002971.1). Primers for these markers were redesigned to amplify significantly shorter PCR products and were combined into 1 multicolor multiplex PCR. Primer sequences for Ms-27 were 5’-HEX-TCTTTATTTCAGGCCGGAGT-3’ and 5’-GAACGACTCATTGAACACACG-3;for Ms-28, 5’-TMR-AGCAAAGAAATGTGAGGATCG-3’and 5’- GCCAAAGGGATATTTTTGTCCTTC-3’; for Ms-34, 5’-FAM-TTCTTCGGTGAGTTGCTGTG-3’ and 5’-GCAATGACTATCAGCGACTCGAA-3’.
†Cycle threshold (Ct) value obtained by using real-time PCR targeting the IS1111a element.
‡NR, no result obtained. A full genotype was obtained only in samples with the highest DNA loads (Ct value <35).

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