Nine Klebsiella pneumoniae isolates showing non-susceptibility to carbapenems were collected from three centres in the north-eastern region of Hungary. The minimum inhibitory concentrations (MICs) of antibiotics were determined by Etest. The putative production of a carbapenemase was tested by the modified Hodge test. The presence of bla (KPC) genes was verified by polymerase chain reaction (PCR) and sequencing. Furthermore, molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). All isolates showed extensively drug-resistant (XDR) phenotype, and of these, eight isolates were highly resistant to colistin. The isolates carried bla (KPC-2), bla (SHV-12), bla (TEM-1) and bla (SHV-11). PFGE analysis of the nine KPC-2-producing Hungarian ST258 K. pneumoniae isolates, two KPC-2-producing Norwegian ST258 isolates and 33 CTX-M-15-producing ST11 isolates revealed the existence of one genetic cluster at an 88% similarity level. The overall results of the PFGE clustering, MLST and the presence of SHV-11 in both ST11 and ST258 suggest that this is the first hyperepidemic clonal complex of multidrug-resistant K. pneumoniae, probably CC258/CC340, possibly undergoing worldwide spread.

Two regions of rpoB associated with rifampin resistance were sequenced in 29 rifampin-resistant (determined by the proportion method) isolates of Mycobacterium tuberculosis obtained from patients from three counties in Hungary. Of the 29 resistant strains, 27 had a mutation in either the 81-bp region (26 strains) or the N-terminal region (1 strain), while the other 2 strains had no mutations in either region. The locations and frequencies of the mutations differed from those previously reported. The most common mutation in this study, D516V, was found in 38% of the Hungarian strains, a frequency 2 to 10 times higher than that found in studies from other countries. These same 29 isolates were also evaluated with the Inno-LiPA Rif. TB test (LiPA), a reverse hybridization assay for the rapid detection of rifampin resistance. Although LiPA detected the presence of an rpoB mutation in 26 of the resistant isolates, the type of mutation could not be determined in 4 isolates because the mutations present were not among those included on the LiPA strip. In addition, a silent mutation in one of the rifampin-susceptible control strains was interpreted as rifampin resistant by LiPA. These findings demonstrate the importance of validating this rapid molecular test by comparison with DNA sequence results in each geographic location before incorporating the test into routine diagnostic work.The recent worldwide increase in the incidence of drugresistant strains of Mycobacterium tuberculosis has highlighted the need for faster and more accurate detection of resistance to rifampin (RMP), one of the most important antituberculosis drugs (17). RMP is most effective in killing actively metabolizing M. tuberculosis, and resistance to RMP often results in high clinical relapse rates (5, 15). Because of the prolonged turnaround time for conventional susceptibility testing, patients infected with drug-resistant tuberculosis may be inadequately treated and thus remain infectious for longer periods than those infected with susceptible strains.Based on collective observations that mutations resulting in an amino acid change within the 81-bp core region of the RNA polymerase ␤-subunit (rpoB) gene are found in more than 96% of RMP-resistant M. tuberculosis strains, several molecular methods have been developed for the rapid (24-to 48-h) detection of mutations in this region (3,10,16,18,24,25,(28)(29)(30). In addition, other studies revealed that mutations associated with RMP resistance can also occur in other regions of the rpoB gene, although less frequently (6,7,23). It has also been shown elsewhere that the information provided by these molecular tests can serve as a molecular epidemiological marker since the relative frequency of the alleles associated with resistance can vary geographically (10,20).Therefore, the aim of the present study was to determine the drug resistance profile of 29 RMP-resistant M. tuberculosis isolates obtained in East Hungary and to detect and identify mutations present in the rpoB gene. Two molecular assays we...

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A nosocomial Hepatitis B virus (HBV) outbreak at a paediatric onco-haematology unit was investigated using molecular biological methods to determine the origin of the infections. The National Reference Laboratory of Hepatitis Viruses received seven HBsAg positive sera from patients and one from the brother of a patient. A fragment of the preS1/preS2/S genes from all samples was amplified, the PCR products were sequenced and a rooted phylogenetic tree was constructed. All nucleotide sequences from the different patients were very similar and 6 of the 8 sequences were identical, suggesting a common origin of the infections. These sequences were closely related to those amplified from a nosocomial HBV epidemic in another hospital in Hungary. The on-scene investigation revealed several malpractices. The two hospital departments had close connections and some of the patients were treated in both institutions. Present report underlines the importance of developing screening protocols for hepatitis viruses and that of the introduction of regular training programs for health care professionals in the field of hospital hygiene.

Vaccination is the main strategy to control severe dehydrating gastroenteritis caused by rotaviruses in early childhood. The availability of new generation rotavirus vaccines has led to an intensification of strain surveillance worldwide, in part, to gauge the impact of the possible vaccine-driven immune selection of wild-type rotavirus strains. In the present study, authors describe the strain prevalence data obtained in 2007, with the involvement of different regions of Hungary. Genomic RNA was extracted from rotavirus-positive stool samples collected mainly from children and then subjected to genotyping using multiplex RT-PCR assay. Type-specific primers targeted G1 to G4, G6, G8 to G10, and G12 VP7 specificities, and P[4], P[6], and P[8] to P[11] VP4 specificities were used. Out of 489 rotavirus-positive specimens, collected from 482 patients, 466 and 474 were successfully G and P typed, respectively, and both G and P type specificities could be assigned for 457 strains. Prevalence data showed the predominance of G4P[8] (31.5%) strains, followed by G1P[8] (28.3%), G2P[4] (19.3%), and G9P[8] (10.2%). Minority strains were G1P[4] (0.4%), G2P[8] (1.3%), G3P[9] (0.2%), G4P[6] (0.7%), G6P[9] (0.4%), G8P[8] (0.2%), G9P[4] (0.2%), G9P[6] (0.8%), and G12P[8] (0.4%). Mixed infections were found in 1.2% of the samples, while 4.9% remained partially or fully non-typified. Our data indicate that the antigen specificities of medically important rotavirus strains identified in this 1-year study are well represented in the vaccines available in the pharmaceutical private market in Hungary. Depending on the vaccination coverage achievable in the forthcoming years, the post-vaccination rotavirus strain surveillance may allow us to gain comprehensive information on the impact of rotavirus vaccines on the prevalence of circulating rotavirus strains.