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Papers by Margaret Sottile

Immunologic studies in pneumococcal disease

PubMed, Jun 1, 1977

Many patients die from pneumococcal disease despite the availability of effective antimicrobial a... more Many patients die from pneumococcal disease despite the availability of effective antimicrobial agents. Immunologic studies including detection, typing, and quantitation of serum pneumococcal capsular polysaccharide (PCP) antigen by counterimmunoelectrophoresis (CIE), quantitation of PCP antibody by radioimmunoassay (RIA), and quantitation of serum complement components C3, C4, and C3PA and serum immunoglobulins IgG, IgM, and IgA by the radial immunodiffusion technique of Mancini were performed with the sera of 18 patients. Five patients died (group I), and 13 survived (group II) pneumococcal infection. Both groups were comparable in age, underlying disease, and leukopenia on admission. All patients of group I and 10 of 13 (77%) of group II patients were bacteremic. Two patients in each group had an extrapulmonary focus infection. PCP antigen was detected in the sera of all group I and nine of 13 group II patients. PCP antigen levels were larger than or equal to 15 microng/ml in four of five group I and two of 13 group II patients (p = 0.022). Levels of antibody to PCP exceeded 100 ng/ml of antibody nitrogen (AbN) in 10 of 12 group II and one of five group I patients (p = 0.027) during the course of illness. All group I patients and three of 12 group II patients had decreased levels of one or more complement components on admission (p less than 0.01). One or more complement components remained decreased until death in four group I patients but returned to normal or elevated levels in all group II patients. No difference in serum immunoglobulin concentrations were found.

Journal of Clinical Microbiology, Sep 1, 1975

The use of counterimmunoelectrophoresis (CIE) as a diagnostic tool in infectious diseases is beco... more The use of counterimmunoelectrophoresis (CIE) as a diagnostic tool in infectious diseases is becoming more widespread. This study was undertaken to determine the possible use of CIE in the more rapid identification of Streptococcus pneumoniae in clinical isolates. Typing sera were obtained from the Statens Seruminstitut, Denmark. Sixty-seven out of 68 pneumococcal isolates that were optochin sensitive and bile soluble were typed by CIE. One isolate was nontypable even after mouse passage. An additional three isolates that were optochin resistant were considered to be pneumococci on the basis of the bile solubility test and their typability by CIE. Seventy-seven a-streptococci were tested for the presence of cross-reacting capsular antigens. Fifty-three a-streptococci showed no cross-reactions using the omniserum. Precipitin bands were obtained with the omniserum with 10 of the isolates, but these did not react with type-specific antisera. However, 14 isolates did react with the type-specific pneumococcal antisera. The sensitivity of the test was increased by sonicating whole-cell suspensions before electrophoresis was carried out. Mueller-Hinton broths were inoculated with presumptive pneumococcal colonies that formed the predominant or only colony type on primary blood agar plates. These cultures required a 4-h incubation period with an initial inoculum of 106 viable organisms/ml before a precipitin band could be detected. Precipitin bands were observed in 54 out of 56 (97%) broth cultures of pneumococci that had been incubated for 4 h at 37 C. These data suggest that CIE could be a useful tool in the identification of S. pneumoniae isolated from clinical specimens.

Staphylococcus Aureus

THE PRODUCTION of coagulase is universally accepted as the most distinctive characteristic of Sta... more THE PRODUCTION of coagulase is universally accepted as the most distinctive characteristic of Staphylococcus aureus. Furthermore, among the numerous extracellular proteins com-monly associated with staphylococcal virulence, coagulase seems to be the most stably associated. Recently we have developed a technique whereby it is readily possible to detect one colony that produces coagulase among 30 OOO coagulase-negative colonies by means of a pour-plate technique (parisi, Baldwin and Sottile, 1973). Coagulase-producing colonies in the agar can be detected by the dense zone of precipitated fibrin around them. This technique makes feasible certain types of genetic studies on production and regulation of coagulase, as well as studies of other aspects of its possible role in staphylococcal disease. MATERIALS AND METHODS Bacteriaf strains. A coagulase-negative mutant was obtained from strain 1-746 after exposure of the culture to 100 pg of N-methyl-N-nitro-N-nitrosoguanidine (Aldrich Chemic...

Pour-Plate MethodfortheDetection of Coagulase Production byStaphylococcus aureus

Apour-plate methodisdescribed forthedetection ofcoagulase production by Staphylococcus aureus.Eit... more Apour-plate methodisdescribed forthedetection ofcoagulase production by Staphylococcus aureus.Either BrainHeartInfusion agaroryeast extract-Trypticase soyagarcontaining swineplasma was thebestmediumforthedetection ofcoagulase bythismethod. Theadvantages ofthismethodanditspotential utilization intheclinical laboratory, infoodmicrobiology, andinspecialized studies withS.aureus arediscussed. Theproduction ofcoagulase isaccepted generally asthemostreliable testforthe identification ofStaphylococcus aureus. Two commonmethods foritsdetection aretheslide test andthetubetest. Theformer, which tests fortheproduction ofboundcoagulase (clumpingfactor), correlates poorly withcoagulase production bythetubemethodandisnot recommended forroutine diagnostic work(4, 6).Penfold (8)andReidandJackson (10) introduced analternate methodofcoagulase testing whereby staphylococci areinoculated ontothesurface ofamediumcontaining plasma.Although thesurface inoculation method hasnotgained wideacceptance, Lackan...

Applied microbiology, 1973

A pour-plate method is described for the detection of coagulase production by Staphylococcus aure... more A pour-plate method is described for the detection of coagulase production by Staphylococcus aureus. Either Brain Heart Infusion agar or yeast extract-Trypticase soy agar containing swine plasma was the best medium for the detection of coagulase by this method. The advantages of this method and its potential utilization in the clinical laboratory, in food microbiology, and in specialized studies with S. aureus are discussed.

Applied Microbiology, 1973

A pour-plate method is described for the detection of coagulase production by Staphylococcus aure... more A pour-plate method is described for the detection of coagulase production by Staphylococcus aureus. Either Brain Heart Infusion agar or yeast extract-Trypticase soy agar containing swine plasma was the best medium for the detection of coagulase by this method. The advantages of this method and its potential utilization in the clinical laboratory, in food microbiology, and in specialized studies with S. aureus are discussed.

American Journal of Medicine, 1978

Study Groups. The study population was derived from patients seen in consultation by the Infectio... more Study Groups. The study population was derived from patients seen in consultation by the Infectious Disease service. Initial specimens were obtained usually shortly after admission, and subsequent serums were collected at weekly intervals throughout the patient's hospitalization. Such follow-up serums were obtained in most, although not all, patients. Serums were assayed for antistaphylococcal antibodies by the CIE method in the following seven groups of subjects.

Journal of Medical Microbiology, 1976

THE PRODUCTION of coagulase is universally accepted as the most distinctive characteristic of Sta... more THE PRODUCTION of coagulase is universally accepted as the most distinctive characteristic of Staphylococcus aureus. Furthermore, among the numerous extracellular proteins commonly associated with staphylococcal virulence, coagulase seems to be the most stably associated. Recently we have developed a technique whereby it is readily possible to detect one colony that produces coagulase among 30 OOO coagulase-negative colonies by means of a pour-plate technique (parisi, Baldwin and Sottile, 1973). Coagulase-producing colonies in the agar can be detected by the dense zone of precipitated fibrin around them. This technique makes feasible certain types of genetic studies on production and regulation of coagulase, as well as studies of other aspects of its possible role in staphylococcal disease.

Immunologic studies in pneumococcal disease

PubMed, Jun 1, 1977

Many patients die from pneumococcal disease despite the availability of effective antimicrobial a... more Many patients die from pneumococcal disease despite the availability of effective antimicrobial agents. Immunologic studies including detection, typing, and quantitation of serum pneumococcal capsular polysaccharide (PCP) antigen by counterimmunoelectrophoresis (CIE), quantitation of PCP antibody by radioimmunoassay (RIA), and quantitation of serum complement components C3, C4, and C3PA and serum immunoglobulins IgG, IgM, and IgA by the radial immunodiffusion technique of Mancini were performed with the sera of 18 patients. Five patients died (group I), and 13 survived (group II) pneumococcal infection. Both groups were comparable in age, underlying disease, and leukopenia on admission. All patients of group I and 10 of 13 (77%) of group II patients were bacteremic. Two patients in each group had an extrapulmonary focus infection. PCP antigen was detected in the sera of all group I and nine of 13 group II patients. PCP antigen levels were larger than or equal to 15 microng/ml in four of five group I and two of 13 group II patients (p = 0.022). Levels of antibody to PCP exceeded 100 ng/ml of antibody nitrogen (AbN) in 10 of 12 group II and one of five group I patients (p = 0.027) during the course of illness. All group I patients and three of 12 group II patients had decreased levels of one or more complement components on admission (p less than 0.01). One or more complement components remained decreased until death in four group I patients but returned to normal or elevated levels in all group II patients. No difference in serum immunoglobulin concentrations were found.

Journal of Clinical Microbiology, Sep 1, 1975

The use of counterimmunoelectrophoresis (CIE) as a diagnostic tool in infectious diseases is beco... more The use of counterimmunoelectrophoresis (CIE) as a diagnostic tool in infectious diseases is becoming more widespread. This study was undertaken to determine the possible use of CIE in the more rapid identification of Streptococcus pneumoniae in clinical isolates. Typing sera were obtained from the Statens Seruminstitut, Denmark. Sixty-seven out of 68 pneumococcal isolates that were optochin sensitive and bile soluble were typed by CIE. One isolate was nontypable even after mouse passage. An additional three isolates that were optochin resistant were considered to be pneumococci on the basis of the bile solubility test and their typability by CIE. Seventy-seven a-streptococci were tested for the presence of cross-reacting capsular antigens. Fifty-three a-streptococci showed no cross-reactions using the omniserum. Precipitin bands were obtained with the omniserum with 10 of the isolates, but these did not react with type-specific antisera. However, 14 isolates did react with the type-specific pneumococcal antisera. The sensitivity of the test was increased by sonicating whole-cell suspensions before electrophoresis was carried out. Mueller-Hinton broths were inoculated with presumptive pneumococcal colonies that formed the predominant or only colony type on primary blood agar plates. These cultures required a 4-h incubation period with an initial inoculum of 106 viable organisms/ml before a precipitin band could be detected. Precipitin bands were observed in 54 out of 56 (97%) broth cultures of pneumococci that had been incubated for 4 h at 37 C. These data suggest that CIE could be a useful tool in the identification of S. pneumoniae isolated from clinical specimens.

Staphylococcus Aureus

THE PRODUCTION of coagulase is universally accepted as the most distinctive characteristic of Sta... more THE PRODUCTION of coagulase is universally accepted as the most distinctive characteristic of Staphylococcus aureus. Furthermore, among the numerous extracellular proteins com-monly associated with staphylococcal virulence, coagulase seems to be the most stably associated. Recently we have developed a technique whereby it is readily possible to detect one colony that produces coagulase among 30 OOO coagulase-negative colonies by means of a pour-plate technique (parisi, Baldwin and Sottile, 1973). Coagulase-producing colonies in the agar can be detected by the dense zone of precipitated fibrin around them. This technique makes feasible certain types of genetic studies on production and regulation of coagulase, as well as studies of other aspects of its possible role in staphylococcal disease. MATERIALS AND METHODS Bacteriaf strains. A coagulase-negative mutant was obtained from strain 1-746 after exposure of the culture to 100 pg of N-methyl-N-nitro-N-nitrosoguanidine (Aldrich Chemic...

Pour-Plate MethodfortheDetection of Coagulase Production byStaphylococcus aureus

Apour-plate methodisdescribed forthedetection ofcoagulase production by Staphylococcus aureus.Eit... more Apour-plate methodisdescribed forthedetection ofcoagulase production by Staphylococcus aureus.Either BrainHeartInfusion agaroryeast extract-Trypticase soyagarcontaining swineplasma was thebestmediumforthedetection ofcoagulase bythismethod. Theadvantages ofthismethodanditspotential utilization intheclinical laboratory, infoodmicrobiology, andinspecialized studies withS.aureus arediscussed. Theproduction ofcoagulase isaccepted generally asthemostreliable testforthe identification ofStaphylococcus aureus. Two commonmethods foritsdetection aretheslide test andthetubetest. Theformer, which tests fortheproduction ofboundcoagulase (clumpingfactor), correlates poorly withcoagulase production bythetubemethodandisnot recommended forroutine diagnostic work(4, 6).Penfold (8)andReidandJackson (10) introduced analternate methodofcoagulase testing whereby staphylococci areinoculated ontothesurface ofamediumcontaining plasma.Although thesurface inoculation method hasnotgained wideacceptance, Lackan...

Applied microbiology, 1973

A pour-plate method is described for the detection of coagulase production by Staphylococcus aure... more A pour-plate method is described for the detection of coagulase production by Staphylococcus aureus. Either Brain Heart Infusion agar or yeast extract-Trypticase soy agar containing swine plasma was the best medium for the detection of coagulase by this method. The advantages of this method and its potential utilization in the clinical laboratory, in food microbiology, and in specialized studies with S. aureus are discussed.

Applied Microbiology, 1973

A pour-plate method is described for the detection of coagulase production by Staphylococcus aure... more A pour-plate method is described for the detection of coagulase production by Staphylococcus aureus. Either Brain Heart Infusion agar or yeast extract-Trypticase soy agar containing swine plasma was the best medium for the detection of coagulase by this method. The advantages of this method and its potential utilization in the clinical laboratory, in food microbiology, and in specialized studies with S. aureus are discussed.

American Journal of Medicine, 1978

Study Groups. The study population was derived from patients seen in consultation by the Infectio... more Study Groups. The study population was derived from patients seen in consultation by the Infectious Disease service. Initial specimens were obtained usually shortly after admission, and subsequent serums were collected at weekly intervals throughout the patient's hospitalization. Such follow-up serums were obtained in most, although not all, patients. Serums were assayed for antistaphylococcal antibodies by the CIE method in the following seven groups of subjects.

Journal of Medical Microbiology, 1976

THE PRODUCTION of coagulase is universally accepted as the most distinctive characteristic of Sta... more THE PRODUCTION of coagulase is universally accepted as the most distinctive characteristic of Staphylococcus aureus. Furthermore, among the numerous extracellular proteins commonly associated with staphylococcal virulence, coagulase seems to be the most stably associated. Recently we have developed a technique whereby it is readily possible to detect one colony that produces coagulase among 30 OOO coagulase-negative colonies by means of a pour-plate technique (parisi, Baldwin and Sottile, 1973). Coagulase-producing colonies in the agar can be detected by the dense zone of precipitated fibrin around them. This technique makes feasible certain types of genetic studies on production and regulation of coagulase, as well as studies of other aspects of its possible role in staphylococcal disease.