Serological typing of coagulase-negative staphylococci using monoclonal antibodies.

This investigation was to determine whether monoclonal antibodies (Mabs) could be used to differentiate coagulase-negative staphylococci (C-NS) at species and strain level. Mabs were produced to four Staphylococcus epidermidis strains, two S. haemolyticus strains, one S. saprophyticus strain and one S. warneri strain. A panel of nine antibodies was tested for species and strain specificity against five type strains and 65 clinical isolates of C-NS by enzyme-linked immunosorbent assay (ELISA). Species specificity was found with Mab D150 produced to one S. haemolyticus strain. Using Mab D150 and Mab D198 in conjunction, identification of 90% of S. haemolyticus isolates to species level was achieved. S. saprophyticus Mab K84 reacted with most other strains of C-NS tested but only three S. haemolyticus strains (16%). This finding provides further evidence that S. haemolyticus possesses different surface determinants to other C-NS which could form the basis of a typing scheme for S. haemolyticus using Mabs D150, D198 and K84.

Prevalence of peritonitis-associated coagulase-negative staphylococci on the skin of continuous ambulatory peritoneal dialysis patients.

The predominance of coagulase-negative staphylococci as normal skin flora is thought to be a factor in their association with episodes of peritonitis in patients undergoing continuous ambulatory peritoneal dialysis. We investigated the prevalence of peritonitis-associated strains on the skin of 28 patients undergoing peritoneal dialysis. Coagulase-negative staphylococci were the most frequently isolated organisms, comprising 47% of peritoneal dialysis fluid isolates and 59% of body site isolates. A total of 142 coagulase-negative staphylococci were speciated, tested for their antimicrobial sensitivity and slime production, and identified by phage typing and plasmid-profile analysis. Staphylococcus epidermidis was the most commonly identified species from both peritoneal dialysis fluid (73%) and body sites (53%). Multiple antibiotic resistance was common, and the greater proportion of isolates were resistant to methicillin; 63.6% of peritoneal dialysis fluid isolates and 61.7% of body-site isolates. S. haemolyticus isolates were significantly more resistant to methicillin than other species. By phage typing and plasmid-profile analysis it was shown that peritonitis was rarely caused by skin-colonizing strains. In only 3 of 14 patients were peritonitis-associated strains isolated as skin colonizers, and no patients developed peritonitis due to organisms previously isolated as skin colonizers.

A study of coagulase-negative staphylococci isolated from clinically significant infections at an Australian teaching hospital.

Some 151 isolates of coagulase-negative staphylococci isolated from patients at an Australian teaching hospital were characterized by biochemical analysis, antibiotic sensitivity patterns and slime production. S. epidermidis was the predominant species (64%) isolated from clinically significant infections, and all S. epidermidis isolates from true bacteremias produced slime. Forty-nine per cent were resistant to methicillin and 61% to gentamicin. S. haemolyticus isolates from clinically significant infections also showed antibiotic resistance and 80% were resistant to more than five antibiotics. The importance of coagulase-negative staphylococci as pathogens in this large teaching hospital was confirmed.

Observations on the resistance to drying of staphylococcal strains.

Death rates have been determined for staphylococcal strains dried on cotton blanket material and stored at room temperature in the dark and in the light. Methicillin-resistant Staphylococcus aureus (MRSA) strains that produced a golden pigment and had a wide distribution within the hospital survived for longer periods than MRSA strains that produced little pigment and had a restricted local distribution. Death rates of methicillin-sensitive strains of S. aureus at day 7 were similar to those of the general epidemic MRSA strains, and there was no significant difference between the death rates at day 7 of the local epidemic MRSA strains and the coagulase-negative strains.

Phage-typing patterns and lysogenicity of methicillin-resistant strains of Staphylococcus aureus from Sydney, Australia, 1965-85.

Methicillin-resistant strains of Staphylococcus aureus isolated at the Royal Prince Alfred Hospital since 1965 were differentiated by phage-typing and by their lysogenic status. Most of these strains were isolated during two periods, 1965-72 and 1976-85. Nearly all of the strains isolated in the first period had one of four phage-typing patterns. Strains with each typing pattern carried two prophages; these eight phages were all different, as characterised by serological grouping and lytic spectrum. Lysogenisation of the non-lysogenic strain 1489 with each of these phages narrowed its phage-typing pattern; the typing pattern of the double lysogens was generally similar to and occasionally identical with that of the host strain that had yielded the pair of phages. In the second period, strains with one of five other phage-typing patterns predominated. Representatives of each of these carried the lysogenic phage C. The first methicillin-resistant strain carrying this phage had been isolated in 1974. The current methicillin-resistant S. aureus strains thus appear to form a distinct group that can be differentiated from those seen in earlier years.

Lysogenicity of methicillin-resistant strains of Staphylococcus aureus.

The lysogenic status of 23 strains of methicillin-resistant Staphylococcus aureus, isolated at the Royal Prince Alfred Hospital, Sydney, since 1980, was studied. Twenty strains, belonging to the four predominant phage types isolated in this hospital, carried the same lysogenic phage which we have designated C. Three other phages were isolated from five strains belonging to phage type 84/85/90. The presence of phage C had little effect on the phage-typing pattern of the strains. Similarly, lysogenization with the other three phages did not result in a significant change in phage-typing patterns. However, when strain 1489, isolated in 1969, was lysogenized with these three phages, there was a change in phage-typing pattern. Lysogenization of this strain with phage 47T resulted in a marked loss of sensitivity to both group-I and group-III phages. The lysogenic status of these methicillin-resistant strains of S. aureus was compared with that of strains isolated between 1967 and 1970. There was no evidence that the strains isolated recently were either related to, or derived from, the earlier ones.

Strain differentiation in methicillin-resistant Staphylococcus aureus.

Three different systems were used to test 236 isolates of methicillin-resistant Staphylococcus aureus in an attempt to ascertain if more than one strain is responsible for the current problem of cross-infection by this organism in N.S.W. hospitals. The biochemical tests used were of little assistance. Phage typing, using the Basic International Set of typing phages at 100 x routine test dilution (RTD), provided evidence of the presence of several different strains. Phage type 83A/85/95/90/88 was the typing pattern of the predominant strain and the nest most frequent group was not typable. These results were often difficult to read. Five new phages were therefore isolated and found to be valuable as they produced easily identifiable patterns at RTD.