ABSTRACT
Ninety-five Neisseria gonorrhoeae organisms isolated in Valencia, Spain, were characterized by antibiotic sensitivity testing, auxotyping, serotyping, plasmid analysis and restriction endonuclease fingerprinting (HindIII digestion). Cluster analysis of the restriction patterns revealed that 31 isolates (32.6%) formed 12 clearly defined clusters. Penicillinase-producing Neisseria gonorrhoeae strains formed four of these groups. Eight groups of gonococcal strains were identified by auxotyping, although 83% of isolates belonged to two auxotypes (Proto, Pro-). Twenty-three different serovars were identified by serotyping. The serovar pattern IB/rop was found in 38% of isolates. A 60% coincidence was found between gonococcal groupings obtained by combination of auxotyping, serotyping and plasmid analysis and those obtained with the restriction enzyme fingerprinting technique. The specificity of enzyme restriction patterns of Neisseria gonorrhoeae is confirmed to be of practical importance in the epidemiologic study of gonorrhoea.
Subject(s)
DNA Fingerprinting , Neisseria gonorrhoeae/classification , Bacterial Typing Techniques , Humans , SpainABSTRACT
OBJECTIVE: To compare the value of different markers and their combinations with the restriction enzyme technique in the differentiations of penicillinase-producing N. gonorrhoeae (PPNG) strains. MATERIALS AND METHODS: 17 PPNG strains isolated from symptomatic, untreated male patients with urethritis were characterised by antibiotic sensitivity testing, auxotyping, serotyping, plasmid profile, and restriction endonuclease fingerprinting (Hind III digestion). Cluster analysis with the method of unweighted pair-group average (UPGMA) linkage was used to calculate similarity or dissimilarity for PPNG strains. MAIN RESULTS: Either auxotyping or plasmid profile alone differentiated three groups of PPNG strains, whereas the combination auxotyping/serotyping identified 10. Although the combination auxotyping/serotyping/plasmid profile and the restriction enzyme technique showed a similar discrimination ability (differentiation of 11 PPNG strains), genomic fingerprinting gave highly specific restriction patterns on individual gonococcal isolates. CONCLUSIONS: The combination of different markers gave more epidemiological information than the use of only one. The sequence of discriminating ability for PPNG strains was: auxotyping/serotyping less than auxotyping/serotyping/plasmid profile less than restriction patterns of genomic DNA.
Subject(s)
DNA Fingerprinting , DNA, Bacterial/analysis , Neisseria gonorrhoeae/genetics , Penicillinase/biosynthesis , Genetic Markers , Genotype , Humans , Male , Neisseria gonorrhoeae/growth & development , Neisseria gonorrhoeae/isolation & purification , Neisseria gonorrhoeae/metabolism , Phenotype , Plasmids , Restriction Mapping , Serotyping , Urethritis/microbiologyABSTRACT
The analysis of electrophoretic profiles of membrane proteins is one of the epidemiological methods of bacterial typing. The profiles of membrane proteins of 95 isolates were studied for valuing their usefulness in the epidemiology of N. gonorrhoeae. The results were compared with the obtained using other characterization methods (auxotyping, serotyping and antimicrobial sensibility). The proteins I and II (PI and PII) showed clear differences between isolates. Only protein-I (PI) with constant molecular weight for each isolate was valid to discriminate between strains. It was observed correlation between serovariety IA and molecular weight of PI 33.6-36 kD, and the serovariety IB with molecular weight 35.5-37 kD. Though it wasn't possible discriminated between the different serovarieties. It was proved a sensibility decrease to penicillin, tetracycline and cloramfenicol in those strains with molecular weight of PI greater than 35.5 kD (serogroup WII/WIII). In the 80% of the isolates considered multiple antibiotic-resistant it was observed a significant increase of the membrane protein dough of 52 kD. All the strains with this protein increased were multiple antibiotic-resistant.
Subject(s)
Membrane Proteins/analysis , Neisseria gonorrhoeae/isolation & purification , Electrophoresis, Polyacrylamide Gel , Epidemiologic Methods , Gonorrhea/epidemiology , Humans , Neisseria gonorrhoeae/chemistry , Spain/epidemiology , Species SpecificityABSTRACT
The screening for respiratory syncytial virus (RSV) in nasopharyngeal secretions with enzyme immunoassay (ELISA) and indirect immunofluorescence (IIF) has been evaluated in infants and young children with acute respiratory infection. Both methods were compared with viral isolation in HEp-2 cells and the investigation of fluorescent foci in cell cultures inoculated by centrifugation. 226 samples were evaluated by IFF, 182 of which were also evaluated by ELISA while 158 were inoculated into cell cultures. 20.35% of samples were positive with IFF and 19.23% with ELISA. Isolation of RSV was obtained in 25 of the samples inoculated into HEp-2 cells (15.8%). The cytopathic effect took a mean of 5.4 days to develop. The investigation of fluorescent foci in centrifugated cultures allowed to detect 76% of positive samples 24 hours after centrifugation and 84% of positive samples 48 hours after it. Considering the viral isolation as the reference method, IIF and ELISA had a 88% and 76% sensitivity, respectively, with very similar specifities (90.2% and 91.7%).
