[Staphylococcal lung diseases in pediatrics; general considerations on staphylococcal infections].
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A 26-year-old female patient with acute myeloid leukaemia was hospitalized for the second cycle of remission induction chemotherapy. While neutropenic she developed progressive pulmonary infiltrate, with Micrococcus spp. cultured from two consecutive bronchoalveolar lavage fluids, resulting in respiratory insufficiency. The patient died after an unsuccessful cardiopulmonary resuscitation. This report of micrococcal pneumonia emphasizes that the pathogenicity of this skin commensal is not limited to infections in tissues surrounding prosthetic devices or indwelling intravenous catheters. Especially in immunocompromised patients, Micrococcus spp. from bronchoalveolar lavage fluids cannot be lightly dismissed as non-pathogenic when pneumonia is considered.
The role of local sequence information in establishing the chromatin structure of the human c-myc upstream region (MUR) was investigated. Adeno-associated virus (AAV)-mediated gene transduction was used to introduce an additional unrearranged copy of the 2.4 kb HindIII-XhoI fragment of the MUR into a novel location in the genome in each of two cloned HeLa cell lines. The AAV-based rep- cap- viral vector SKMA used to transduce the MUR retained only 1.4 kb (24%) of the AAV genome and could accommodate inserts as large as 2.4 kb. SKMA was capable of infecting HeLa cells and integrating into the host genome at single copy number. Integration may have occurred at a preferred site in the HeLa genome, but this site was apparently distinct from the previously identified preferred AAV integration site on human chromosome 19. Indirect end-labelling was used to map DNase I and micrococcal nuclease (MNase) cleavage sites over the transduced c-myc sequences and the endogenous c-myc loci in infected HeLa cells. A similarly ordered chromatin domain, extending 5' from c-myc promoter P0, was found to exist at the transduced c-myc locus in each clone. The position and relative sensitivity of 13 MNase cleavage sites and five DNase I hypersensitive sites, originally identified at the endogenous MUR in non-transduced cells, were shown to be conserved when this DNA was moved to a new chromosome site. A conserved DNase I hypersensitive site also was mapped to the region between the left AAV terminal repeat and AAV promoter P5. These results suggest that the information required to establish the particular chromatin structure of the MUR resides within the local DNA sequence of that region.
The purpose of the study was the molecular characterization of methicillin-resistant Staphylococcus aureus (MRSA) isolates cultured from patients treated in seven wards of a university hospital in Lublin, Poland, over a 14-month period. Eleven nosocomial MRSA isolates were analyzed. Phenotypic identification of the isolates as MRSA was confirmed by the detection of the nuc and mecA genes using a multiplex PCR assay. The MRSA isolates were further characterized by pulsed-field gel electrophoresis, 16S-23S rRNA spacer length polymorphism analysis, and the simplex and multiplex SCCmec PCR assays. The MRSA isolates were found to be multiresistant: in addition to resistance to beta-lactam agents, they demonstrated resistance to ciprofloxacin, tetracycline, erythromycin, and gentamicin. The MRSA isolates were genetically identical and shared common pulsed-field gel electrophoresis profiles and 16S-23S rRNA spacer length polymorphism profiles. The PCR-based method revealed that the profile of the Lublin clone was identical to that of the Brazilian pandemic MRSA isolates. By SCCmec typing, all MRSA isolates harbored the C variant of the SCCmec type III that differed from the typical SCCmec type III pattern by the lack of locus F (414 bp). The results of this study indicate the spread of a single, multiresistant, MRSA clone in various wards of a university hospital over a 14-month period. The SCCmec structure harbored by the Lublin clone has previously been identified among Polish MRSA isolates representing the HoMRSA-Pol1 clone. The data from this study indicate that the Lublin MRSA clone is most probably genetically related to the HoMRSA-Pol1 clone. Moreover, this latter clone belongs to ST239, the same sequence type as the Hungarian and Brazilian pandemic MRSA isolates.
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An ELISA method based on monodisperse magnetic beads was developed for the detection of Staphylococcus aureus thermostable nuclease (TNase) in composite milk, wherein S. aureus TNase is captured by magnetic beads coated with monoclonal antibodies directed against TNase and subsequently detected by an enzyme-labelled MAb against the same antigen. Sensitivity of the test was approximately 1 ng TNase, which corresponds to the amount of TNase produced and secreted by approximately 10(5) S. aureus per ml. The Immuno Magnetic Separation (IMS)-ELISA detected TNase in samples from which no S. aureus could be demonstrated on culture. The total test time is 3 h and can be performed either on preserved or fresh milk. The method may be automated.
The nuclease-neutralisation test was evaluated as a means of identifying nuclease-positive staphylococci isolated from different animals. The test identified 510 of 520 strains (98%) of staphylococci characterised by biochemical testing.
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