Differentiation of enterococci from other group D streptococci by means of a specific monoclonal antibody
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We report on the specificity of a monoclonal antibody which reacts with autoclaved extracts of four species of enterococci but does not react to the same extent with similar extracts from two non-enterococcal group D streptococci. The monoclonal antibody also reacts specifically with purified lipoteichoic acid from Streptococcus faecalis but not significantly with purified lipoteichoic acid from the non-enterococcal species Streptococcus bovis and Streptococcus equinus. The specific antigen detected with this antibody could correlate with the definition of the enterococcus sub-group of the streptococci which would provide further evidence that this sub-group is taxonomically distinct from the other group D streptococci.A 4-h method was devised to differentiate the non-beta-hemolytic streptococci into three categories: enterococci, group D nonenterococci, and viridans streptococci. All of the Streptococcus faecalis, 90% of the Streptococcus faecium (enterococci), and 96% of the Streptococcus bovis biotype I (group D nonenterococci) cultures were correctly identified by the 4-h method. The less commonly isolated group D cultures had lower rates of correct identification by this method. None of the viridans streptococci was identified incorrectly.
Streptococcus bovis
Viridans streptococci
Enterococcus faecalis
Enterococcus faecium
Identification
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Whole cell protein profiles were resolved for Streptococcus equisimilis (group C) and large colony human biotype beta-haemolytic group G streptococci by the use of one dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Strains of S. equisimilis (27 in toto) were distributed among eight patterns designated A to H. Pattern A represented 48.2% of the latter isolates. Strains of group G streptococci (59 in toto) were distributed among sixteen patterns designated 1-16, and there were no predominant patterns which represented more than 20% of all strains. Profiles were reproducible, not susceptible to strain passage, but susceptible to variation in growth media. Considerable homology was observed among bacteria in either Lancefield group.
Group A
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Homology
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Similar α-(1→6) linkage-rich, soluble, extracellular glucans have been isolated from six strains of two genetically distinct groups of Streptococcus sanguis and three strains of Streptococcus mitior.
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Identification of a streptococcal penicillin-binding protein that reacts very slowly with penicillin
Penicillin-binding protein (PBP) 5 of Streptococcus faecium ATCC 9790 has an unusually low affinity for penicillin (50% binding occurred at a penicillin level of 8 micrograms/ml after 60 min of incubation, and the protein only became labeled after 20 min of incubation with high concentrations of radioactive penicillin). PBPs with similar properties are carried by strains of Streptococcus durans, Streptococcus faecalis, and Streptococcus lactis but not by strains of groups A, B, C, and G streptococci or Streptococcus pneumoniae. The strains carrying the slow-reacting PBP demonstrated a sensitivity to penicillin that was several hundred times lower than that of strains not carrying it. Spontaneous mutants with minimal inhibitory concentrations of penicillin of 20, 40, and 80 micrograms/ml were isolated from S. faecium ATCC 9790. They all showed a dramatic increase in the amount of slow-reacting PBP produced. Mutants with increased penicillin resistance were also isolated from wild-type strains of S. durans, S. faecalis, and S. faecium. All of them carried a greater amount of the slow-reacting PBP than that carried by the parent. Finally, it was found that resistant S. faecium ATCC 9790 mutants grew normally in the presence of penicillin concentrations that were far above that saturating all PBPs except PBP 5. Cell growth was, on the contrary, inhibited by a penicillin concentration that saturated the slow-reacting PBP by 90%. This penicillin dose was equal to the minimal inhibitory concentration.
Penicillin binding proteins
Streptococcus bovis
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The properties of the lactate dehydrogenases, percent guanine plus cytosine in the deoxyribonucleic acid (DNA), and DNA/DNA hybridization studies have shown that three strains of group N streptococci do not belong to either Streptococcus lactis or Streptococcus cremoris. The biochemical properties of the three strains were published about 25 years ago, and at that time the strains were not assigned to any species. The three strains are here identified as members of Streptococcus raffinolactis Orla-Jensen and Hansen.
Cytosine
Streptococcus mitis
Streptococcus bovis
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We report on the specificity of a monoclonal antibody which reacts with autoclaved extracts of four species of enterococci but does not react to the same extent with similar extracts from two non-enterococcal group D streptococci. The monoclonal antibody also reacts specifically with purified lipoteichoic acid from Streptococcus faecalis but not significantly with purified lipoteichoic acid from the non-enterococcal species Streptococcus bovis and Streptococcus equinus. The specific antigen detected with this antibody could correlate with the definition of the enterococcus sub-group of the streptococci which would provide further evidence that this sub-group is taxonomically distinct from the other group D streptococci.
Lipoteichoic acid
Enterococcus faecalis
Streptococcus bovis
Group A
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We report on the specificity of a monoclonal antibody which reacts with autoclaved extracts of four species of enterococci but does not react to the same extent with similar extracts from two non-enterococcal group D streptococci. The monoclonal antibody also reacts specifically with purified lipoteichoic acid from Streptococcus faecalis but not significantly with purified lipoteichoic acid from the non-enterococcal species Streptococcus bovis and Streptococcus equinus. The specific antigen detected with this antibody could correlate with the definition of the enterococcus sub-group of the streptococci which would provide further evidence that this sub-group is taxonomically distinct from the other group D streptococci.
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The results of deoxyribonucleic acid-deoxyribonucleic acid and deoxyribonucleic acid-ribosomal ribonucleic acid hybridization studies demonstrated that Streptococcus faecalis and Streptococcus faecium are distantly related to the non-enterococcal streptococci (Streptococcus bovis and Streptococcus equinus) of serological group D and to other streptococci. On the basis of our results and those of previous studies, we propose that S . faecalis and S . faecium be transferred to the genus Enterococcus (ex Thiercelin and Jouhaud) nom. rev. as Enterococcus faecalis (Andrewes and Horder) comb. nov. and Enterococcus faecium (Orla-Jensen) comb. nov., respectively. A description of the genus Enterococcus nom. rev. and emended descriptions of E. faecalis and E. faecium are given.
Enterococcus faecalis
Streptococcus bovis
Enterococcus faecium
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Streptococcus bovis
Streptococcus equi
Streptococcus sanguinis
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Human anti-murine antibodies (HAMA) can be found in serum of many patients who have received murine monoclonal antibodies for diagnosis or therapy. These antibodies are known to give false positive results in sandwich-type assays (e.g. ELISA or RIA). This interference problem will increase in the future as more patients are treated with murine monoclonal antibodies in vivo. HAMA can also be found in sera from patients that has not been treated with monoclonal antibodies. In this work we have studied the interference of HAMA in sandwich ELISAs containing antibodies from different species. HAMA, present in the sample, may react both with the capture antibody and the detection antibody in these assays to give a false positive reaction. HAMA did not react with chicken IgG, and if one of (or both) the capture and detection antibody was of avian origin, the interference of HAMA in sandwich assays could be avoided.
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