Constantly expressed genes are used as internal controls in relative quantification studies. Suitable internal controls for such studies have not yet been defined for Pseudomonas aeruginosa. In this study, the genes ampC, fabD, proC, pbp-2, rpoD and rpoS of P. aeruginosa were compared in terms of expression stability by real-time quantitative RT-PCR. A total of 23 strains with diverse resistance phenotypes were studied. Stability of expression among the housekeeping genes was assessed on the basis of correlation coefficients, with the best-correlated pair accepted as being the most stable one. Eventually, proC and rpoD formed the most stable pair (r ¼ 0·958; P , 0·001). Next, in four ciprofloxacin-selected nfxC-like mutants, levels of oprD, oprM and oprN mRNA were compared with those of their wild-type counterparts. The comparison was made after correcting the raw values by the geometric mean of the internal control genes proC and rpoD. The level of oprN mRNA was significantly up-regulated, while the oprD gene was down-regulated (although this difference was statistically insignificant), in the mutants. This expression pattern was consistent with that of the expected expression profile of nfxC-type mutants; this experiment therefore lends further support to the use of proC and rpoD genes simultaneously as internal controls for such studies.

A Crimean-Congo haemorrhagic fever (CCHF) outbreak emerged from 2001 to 2003 in the Middle Anatolia region of Turkey. This study describes the clinical characteristics and outcome features of CCHF patients admitted to four tertiary care hospitals in Turkey. Definitive diagnosis was based on the detection of CCHF virus-specific IgM by ELISA or of genomic segments of the CCHF virus by RT-PCR. Related data were collected by a retrospective chart review. Hospital costs were extracted from the final discharge bills. Univariate and multivariate analyses were conducted to determine the independent predictors of mortality. CCHF virus-specific antibodies or genomic segments were detected in the sera of 99 cases. Seven cases that were treated with ribavirin were excluded from the study. Cases were mostly farmers (83 cases, 90 %), and 60 % had a tick-bite history before the onset of fever. Impaired consciousness and splenomegaly were independent predictors of a fatal outcome.