Inexpensive 4-hour micro-agar dilution susceptibility determination method.

Using a micro-agar dilution (MAD) method in which microscope slides are covered with a thin film of agar, and MICs are read microscopically after a 4-h incubation, 18 antibiotics were tested against 29 to 32 microorganisms each. Identical MICs were obtained for microscopic MAD MICs performed in duplicate in 87.1% of the antibiotic-microorganism combinations, and 97.9% were identical within one dilution. When read macroscopically after an 18-h incubation, identical duplicate MICs were obtained in 86.8% of the cases, and 98.4% were identical within one dilution. Using agar dilution as the "gold standard," the correlation obtained with MAD slides read microscopically at 4 h was 94.3%, and macroscopic correlation at 18 h was 97.6%. The correlation of MAD slides with agar dilution for the groups of microorganisms most frequently used was as follows (microscopic/macroscopic): Staphylococcus aureus 96%/98%; Streptococcaceae 97%/98%; Enterobacteriaceae 98%/99%; and Pseudomonadaceae 95%/98%. At the present rate of exchange (fl 1.60 = $1.00f1p4he cost of a MAD slide, including labor, is $1.28 (20 microorganisms tested) or $0.06 per microorganism-antibiotic combination tested. This method is easy to perform, rapid, and inexpensive. It is suitable for use in routine and research laboratories.

Demonstration of circulating pneumococcal immunoglobulin G immune complexes in patients with community-acquired pneumonia by means of an enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay was developed for quantitation of circulating immune complexes (CICs) containing specific antipneumococcal immunoglobulin G (IgG). These CICs were detected in 17 (85%) of 20 patients with bacteremic pneumococcal pneumonia, 4 (36.4%) of 11 patients with probable pneumococcal pneumonia, 3 (16.7%) of 18 patients with pneumonia of other (nonpneumococcal) etiology, and 13 (41.9%) of 31 patients with pneumonia of unknown etiology. There was no correlation between CICs and serum IgG antibody levels. Pneumococcal capsular antigen was demonstrated in dissociated CICs by latex agglutination.

Antigen detection in oropharyngeal secretions for rapid diagnosis of pneumococcal pneumonia.

To determine the value of detection of antigen in the oropharynx in the diagnosis of pneumococcal pneumonia, oropharyngeal secretions were cultured for the presence of Streptococcus pneumoniae and tested for the presence of pneumococcal antigen. Sputum (if available) collected on the same day was also investigated for the presence of antigen. Detection of pneumococcal antigen was found to be directly related to the severity of pneumococcal carriership or infection (p < 0.0001) and was not related to culture results. Patients with pneumococcal pneumonia had the highest antigen detection rate (38%), followed by patients with pneumonia of unknown etiology (32%) and patients with an acute lower respiratory tract infection due to Streptococcus pneumoniae (20%). Pneumococcal carriers had a detection rate of only 9%. Antigen could be detected in only one patient of the control groups. Although antigen detection in sputum was superior to that in oropharyngeal secretions, concordant results were obtained in 8 (40%) and 6 (36%) patients with pneumococcal pneumonia and pneumonia of unknown etiology respectively. The results strongly suggest that pneumococcal carriage seldom leads to a detectable level of antigen, and that antigen detection in the oropharynx appears to be of additive value in the diagnosis of pneumococcal pneumonia.

Rapid detection of pneumococcal antigen in pleural fluid of patients with community acquired pneumonia.

BACKGROUND: Detection of pneumococcal antigen may help to increase the rate of diagnosis of pneumococcal pneumonia. This study was designed to determine the value of rapid detection of pneumococcal antigen in pleural fluid from patients with community acquired pneumonia. METHODS: Thoracentesis was performed in patients suspected of having empyema and in patients with pneumonia of unknown aetiology. Pneumococcal capsular antigen was detected by latex agglutination and this method was compared with Gram staining and culture, specimens of pleural fluid being examined in parallel by the three methods. RESULTS: Pleural fluid was radiographically identified in 63 of 135 patients with community acquired pneumonia. In nine of 45 patients with pneumococcal pneumonia and pleural fluid pneumococci were identified by Gram stain in two and by culture in one specimen of pleural fluid, whereas antigen was detected in eight of these specimens. In 12 of 33 patients with pneumonia of other known aetiology only one pleural fluid specimen was antigen positive, providing a specificity of 92% for this test. Pleural fluid obtained from 12 of 58 patients with pneumonia of unknown aetiology yielded detectable antigen in seven cases. CONCLUSIONS: Detection of pneumococcal antigen by latex agglutination in pleural fluid may yield important and rapid information in patients with community acquired pneumonia.

The role of antigen detection in pneumococcal carriers: a comparison between cultures and capsular antigen detection in upper respiratory tract secretions.

During the winter season upper respiratory tract secretions from 166 patients with stable chronic obstructive pulmonary disease (COPD) or asthma were simultaneously cultured for Streptococcus pneumoniae and tested for pneumococcal capsular antigen. Latex agglutination was employed to investigate the effect of pneumococcal carriership on pneumococcal capsular antigen detection in upper respiratory tract secretions. All specimens originating from the oropharynx, nasopharynx and saliva were both cultured and investigated in parallel for the presence of antigen. The recovery of pneumococci from the different areas was unequally distributed (oropharynx 29%, nasopharynx 8%, and saliva 16%), with the highest isolation rate from the oropharynx alone. Only 4 (3%) of the oropharyngeal swabs, 1 (1%) of the nasopharyngeal swabs and 14 (9%) of the saliva specimens yielded both pneumococcal antigen and a positive culture for S. pneumoniae. A further 9 (6%) of the oropharyngeal swabs, 5 (3%) of the nasopharyngeal swabs, and 50 (33%) of the saliva specimens were antigen positive only, with no pneumococci isolated on culture. It is speculated that these reactions were due to cross-reacting microorganisms (especially alpha-haemolytic streptococci) present in saliva and contaminating the oropharynx and the nasopharynx. Quantitative cultures of 9 oropharyngeal swabs yielded S. pneumoniae in concentrations too low to be detectable by latex agglutination. The study indicates that there is a poor relation between pneumococcal colonization and antigen detection in the oropharynx and nasopharynx. Antigen present in these secretions is probably not an important disrupting factor by contamination when detecting pneumococcal antigen in washed sputum. The false positive antigen results in saliva are probably due to cross-reactions with alpha-haemolytic streptococci.

Detection of pneumococcal capsular antigen in the presence of penicillin in vitro.

Eight strains of Streptococcus pneumoniae were tested in vitro for their ability to produce capsular antigen in the presence of penicillin. It was found that, provided 10(6) to 10(7) pneumococci/ml were present, capsular antigen could be detected during the 72 h in which the experiment was conducted, irrespective of whether penicillin was added at 0 h or 8 h, and even when no viable pneumococci remained. When fewer pneumococci were present, capsular antigen could not be detected at any time in the presence of penicillin. Control cultures, without penicillin, yielded detectable capsular antigen only when the threshold value of 10(6)-10(7) pneumococci/ml was reached. It is concluded that the presence of penicillin does not influence the detection of pneumococcal capsular antigen, but demonstration of this antigen is totally dependent on the number of pneumococci present.

Pneumococcal antigen persistence in sputum from patients with community-acquired pneumonia.

The purpose of this study was to establish the diagnostic value of pneumococcal capsular antigen by comparing this with the results of Gram stain and culture in representative and nonrepresentative sputa during follow-up in patients with community-acquired pneumonia. Antigen was detected by a latex particle agglutination test. At the time of hospital admission, antigen was detected in 17 representative sputum specimens from 30 patients with pneumococcal pneumonia, which was comparable to the results of Gram stain and culture. In five additional patients, antigen was demonstrated in nonrepresentative specimens. During follow-up under antibiotic treatment, this number increased by six: three patients with representative and three patients with nonrepresentative sputum, respectively. Two of the 22 patients with pneumonia of other known cause had an antigen-positive sputum on admission and in another two patients, sputum antigen was detected during follow-up. Ten of 34 patients with pneumonia of unknown cause had detectable antigen in representative or nonrepresentative sputum on admission. During follow-up, antigen was detected in sputa of an additional seven patients. There was no difference in duration of antigen persistence between patients with pneumococcal pneumonia and pneumonia of unknown cause. It was observed that the first antigen-positive sputum specimen was always detected within the first five days of the hospital stay. We conclude that antigen detection in both representative and nonrepresentative sputum specimens at the time of hospital admission and during follow-up is of additional value for the diagnosis of pneumococcal pneumonia. It markedly increases the number of patients with pneumococcal pneumonia detected, who would otherwise be considered to have pneumonia of unknown cause. However, antigen-positive results should be interpreted carefully, especially in those pneumonia patients with chronic bronchitis, because detectable antigen may be caused by pneumococcal carriership of the lower respiratory tract.

Minimum number of pneumococci required for capsular antigen to be detectable by latex agglutination.

Forty-eight strains of Streptococcus pneumoniae were tested in vitro to determine the minimum number required for pneumococcal capsular antigen to be detectable by latex agglutination. It was found that 10(6) to 10(7) microorganisms per ml were needed and that antigen remained detectable even when viable pneumococci could no longer be demonstrated.

Pneumococcal capsular antigen detection and pneumococcal serology in patients with community acquired pneumonia.

BACKGROUND: Methods to determine the microbial cause of community acquired pneumonia include detection of pneumococcal antigen and measurement of pneumococcal capsular antibody response. Their usefulness compared with conventional microbiological techniques was investigated in patients with pneumonia, some of whom had been treated with antibiotics. METHODS: Pneumococcal capsular antigen was detected by latex agglutination in sputum and the results compared prospectively with results of conventional microbiological techniques in 90 patients with community acquired pneumonia. Serum, urine, and pleural fluid samples were also tested for antigen. Serum pneumococcal capsular antibody titres were measured. RESULTS: A diagnosis was established by conventional microbiological techniques in 53 patients, 30 of whom had pneumococcal pneumonia. The sensitivity of antigen detection in first day sputum specimens (n = 18) in those with pneumococcal pneumonia was 94%; antigen was present in 23 of the 27 patients who produced representative sputum on admission and during follow up. The specificity of antigen detection in sputum in patients with non-pneumococcal pneumonia and lung infarction was 87%. Antigen was present in 12 of 25 patients with pneumonia of unknown aetiology who produced representative sputum. Antigen was rarely detected in serum and urine, but was present in pleural fluid in three of four patients with pneumococcal pneumonia and in all four patients with pneumonia of unknown aetiology. Pneumococcal antigen remained detectable in patients treated with antibiotics. Pneumococcal capsular antibody detection was as specific (85%) as antigen detection, but had a lower sensitivity (50%). CONCLUSION: Pneumococcal antigen detection in sputum or pleural fluid is of value in making a rapid diagnosis and provides an additional diagnostic result in patients with pneumococcal pneumonia, especially those receiving antibiotic treatment.

Effect of washing sputum on detection of pneumococcal capsular antigen.

The effect of washing of sputum on detection of pneumococcal capsular antigen was investigated. A total of 357 sputa from 104 patients was tested. Antigen could be detected in 164 (46%) of the sputa in both the washed and unwashed portions, and could not be detected in either portion in a further 180 (50%) sputa. Four (1%) of the sputa agglutinated in the negative control, and were considered to be auto-agglutinating. In 9 (3%) sputa antigen could be detected in the unwashed portion, but not in the washed portion. There were no specimens in which antigen could be detected in the washed portion only. These data indicate that pneumococcal capsular antigen can be detected as reliably in washed sputum as in unwashed sputum.

Incidence of postextraction bacteremia under penicillin cover in children with cardiac disease.

To assess the efficacy of penicillin prophylaxis as recommended by the American Heart Association to prevent the onset of bacterial endocarditis, the incidence of postextraction bacteremia was determined in 82 children with cardiac disease who were receiving prophylactic penicillin. Aerobic and anaerobic blood cultures were taken five minutes after dental extraction, as was a blood sample to assay the serum penicillin concentration. The incidence of postextraction bacteremia was 21%. Streptococcal species accounted for half of the number of aerobes isolated. Of the isolated microorganisms, 16% were strict anaerobes. Susceptibility testing of the isolates showed that 24 penicillin-sensitive and eight penicillin-resistant microorganisms caused the bacteremia. There was neither a significant difference between the serum penicillin concentrations of children with and without bacteremia, nor between the serum penicillin concentrations of children with bacteremia due to penicillin-sensitive microorganisms and children with bacteremia due to penicillin-resistant microorganisms. It is concluded that the occurrence of postextraction bacteremia is not prevented by penicillin prophylaxis and that the serum penicillin concentration at extraction is not the discriminating factor in preventing this bacteremia. None of the children developed bacterial endocarditis. It is speculated that mechanisms not thoroughly studied are involved in the prevention of bacterial endocarditis after dental extraction.