Comparison of the BacT/Alert PF pediatric FAN blood culture bottle with the standard pediatric blood culture bottle, the Pedi-BacT.

The performance of the BacT/Alert PF (Organon-Teknika Corp., Durham, N.C.), a new nonvented pediatric FAN blood culture bottle, was compared to that of the original pediatric bottle, the Pedi-BacT, with matched aerobic cultures obtained from two separate facilities. A total of 244 clinically significant isolates were recovered from 4,015 compliant pairs. Among the positive cultures, 170 (70%) isolates were detected in both the BacT/Alert PF and the Pedi-BacT bottles, while 47 (19%) isolates were recovered in the BacT/Alert PF bottle only and 27 (11%) isolates were recovered in the Pedi-BacT bottle only. Although isolation of specific microorganisms was comparable for the two bottles, the total number of organisms recovered by the BacT/Alert PF was greater than that by the Pedi-BacT (P = 0.0272). In addition, more organisms were recovered by the BacT/Alert PF bottle from the blood of patients receiving antimicrobial therapy (P = 0.0180). Overall time to detection was similar for the two bottles; however, a significantly decreased mean time to detection was recorded for yeast from the BacT/Alert PF bottle (22.9 h; P = 0.0001) and staphylococci from the Pedi-BacT bottle (22.5 h; P = 0.0056). One false-negative culture and five false-positive cultures occurred with the Pedi-BacT bottle, compared to one false-positive culture with the BacT/Alert PF bottle. The BacT/Alert PF bottle is a reliable blood culture bottle for pediatric blood culture specimens and may offer improved recovery of microbes from patients on antimicrobial therapy. The use of the nonvented bottle will both facilitate bottle processing and decrease expenditures for materials due to the elimination of the venting needles required for the original vented bottles.

Disseminated Microascus cirrosus infection in pediatric bone marrow transplant recipient.

Microascus cirrosus Curzi and its associated anamorphic state, Scopulariopsis, were recovered from the cutaneous lesion of a 12-year-old male who had undergone an autologous bone marrow transplantation for acute myelogenous leukemia. Histopathology sections from the biopsied lesion demonstrated septate hyphae consistent with a fungal etiology. Radiographic studies of the lungs subsequent to progression of the lesion revealed a consolidation in the right upper lobe suggesting a primary focus of infection. While M. cirrosus is fairly abundant in nature and widely distributed in stored grains in North America, this is, to our knowledge, the first reported human infection by this species. Salient characteristics for the identification of this dematiaceous ascomycete, M. cirrosus, will be presented.

Comparison of three commercial MIC systems, E test, fastidious antimicrobial susceptibility panel, and FOX fastidious panel, for confirmation of penicillin and cephalosporin resistance in Streptococcus pneumoniae.

The performances of three commercial broth microdilution MIC assays adapted for use with fastidious organisms--the E test (ET), Fastidious Antimicrobial Susceptibility panel (FAS), and FOX Fastidious panel (FOX)--were compared with a MIC using Mueller-Hinton broth with 5% lysed horse blood (MHLHB) to confirm penicillin and cephalosporin resistance in clinical isolates of Streptococcus pneumoniae. Of the isolates screened for penicillin resistance, 5 (12.8%) were categorized as susceptible, 16 (41.0%) were categorized as intermediate, and 18 (46.2%) were categorized as resistant by MHLHB. Only the isolates exhibiting intermediate-to-resistant MICs were included in the comparison. Agreement within +/- 1 log2 dilution was found in 91, 21, and 76% of the ET, FAS, and FOX MICs, respectively, compared with the MHLHB MIC. No very major or major discrepancies occurred with the ET or FOX; however, two very major interpretive errors occurred with the FAS. Agreement between the ET and MHLHB for cefotaxime, ceftriaxone, and cefuroxime was 88, 85, and 100%, respectively. Less than 50% of cephalosporin MICs categorized as > 0.5 microgram/ml by MHLHB were detected by FAS or FOX. Of the methods compared, the ET was the most reliable alternative for susceptibility testing of pneumococci.

Comparison of the BacT/Alert pediatric blood culture system, Pedi-BacT, with conventional culture using the 20-milliliter Becton-Dickinson supplemented peptone broth tube.

The performance of the Pedi-BacT system, the BacT/Alert (Organon Teknika Corp., Durham, N.C.) pediatric blood culture bottle, was compared with that of a conventional 20-ml supplemented peptone broth tube (Becton-Dickinson Corp., Cockeysville, Md.) (BD system) in matched aerobic cultures. The tubes of the BD system were visually examined daily for 7 days and were subcultured during the first 24 h of incubation. Pedi-BacT cultures were mechanically agitated and continuously monitored for growth by the instrument. Of the 6,628 compliant pairs, 331 (5.0%) were positive in both systems, 220 (3.3%) were positive in the Pedi-BacT system only, and 170 (2.6%) were positive in the BD system only. One (0.02%) false-negative culture and 15 (0.2%) false-positive cultures occurred with the Pedi-BacT system while 20 (0.3%) false-negative cultures and 35 (0.5%) false-positive cultures occurred with the BD system. Of 288 clinically significant organisms detected in matched pairs from which a single isolate was recovered, 176 (61%) were recovered from both systems, 83 (29%) were recovered from the Pedi-BacT system only (P < 0.0001), and 29 (10%) were recovered from the BD system only. Members of the family Enterobacteriaceae (P < 0.01), miscellaneous nonfermenters (P < 0.05), and Candida spp. (P < 0.01) were isolated more frequently in the Pedi-BacT system than in the BD system. No significant difference in recovery of other organisms was found between the systems. The average time to detection for the Pedi-BacT system ranged from 11.5 h for streptococci to 29.7 h for enterococci, while that for the BD system ranged from 20.3 h for streptococci to 66.4 h for some nonfermenters. The BacT/Alert system is a reliable, labor-saving alternative to conventional blood culture methods.

Comparison of direct fluorescent antibody, acridine orange, wet mount, and culture for detection of Trichomonas vaginalis in women attending a public sexually transmitted diseases clinic.

To define the performance characteristics of two newer tests for Trichomonas vaginalis (TV), the authors compared direct fluorescent antibody (DFA) (mixed monoclonal antibody, Integrated Diagnostics, Inc, Berkeley, CA) and acridine orange (AO) tests to standard wet mount (WM) preparations and culture (modified Diamond medium) of vaginal wash specimens in consecutively examined women presenting to a public sexually transmitted diseases (STD) clinic. Cultures for Neisseria gonorrhoeae (GC), Chlamydia trachomatis (CT), and yeast were also performed on all patients. Of 104 women, 59 (57%) were infected with one or more pathogens. Trichomonas vaginalis was detected by WM and/or culture in 38 (37%) women and was the most prevalent infection. Of the 38 patients with TV, 95% were detected by culture, 83% by DFA, 66% by AO, and 66% by WM. An additional patient was DFA positive but negative for TV by all other methods. The sensitivity of DFA was superior to AO and WM in women with TV infection alone (96% compared to 67% and 53%, respectively). It was comparable to AO and WM in women with multiple infections (67% compared to 53% and 73%). Even in the presence of other pathogens, DFA appears to be a reasonable alternative to culture for detection of TV. In addition, DFA is rapid, easy to perform, and relatively inexpensive.

Primary pulmonary infection caused by Mycobacterium terrae complex.

The Mycobacterium terrae complex, consisting of three saprophytic species, M. terrae, M. nonchromogenicum, and M. triviale, rarely causes human disease. Only six cases of respiratory infection involving the complex have been documented worldwide. A case of primary pulmonary disease in a previously healthy young woman caused by M. terrae complex is described.

Rapid and overnight microdilution antibiotic susceptibility testing with the Sensititre Breakpoint Autoreader system.

The Sensititre Breakpoint Autoreader system (SBAS) is a broth microdilution method with one to three concentrations of each antibiotic and innovative fluorescence technology to define inhibitory endpoints. We tested 248 gram-negative bacilli and 80 gram-positive cocci using both the rapid (5 h) and overnight (18 h) SBAS procedures. Inhibitory endpoints were also determined by visual inspection of the microdilution trays after 18 h of incubation. SBAS results were compared with those obtained by a standardized disk diffusion (SDD) procedure. Agreement between the rapid SBAS and SDD results for all antibiotic-organism combinations was found in 3,730 of 4,571 (81.6%) tests, with 3.9% very major, 6.5% major, and 7.9% minor discrepancies noted. Data analysis by organism group revealed 86.8, 57.3, 71.4, and 62.3% agreement for members of the family Enterobacteriaceae, Pseudomonas spp., staphylococci, and enterococci, respectively. The results of the overnight SBAS and SDD agreed in 4,154 of 4,654 (89.2%) tests, with 2.3% very major, 1.3% major, and 7.1% minor discrepancies recorded. Concordance was noted in 90.4, 78.1, 90.6, and 83.3% of the comparisons for the members of the Enterobacteriaceae, Pseudomonas spp., staphylococci, and enterococci, respectively. The inhibitory endpoints determined with the Autoreader were as reliable as those determined by visual inspection after 18 h of incubation.

Evaluation of three types of cell culture for recovery of adenovirus from clinical specimens.

The performance characteristics of HEK, HDF, and MK cells for adenovirus isolation were examined for eye and respiratory tract specimens. HEK cells were superior to HDF and MK cells in terms of both speed of virus detection and sensitivity.

A study of anti-group A streptococcal monoclonal antibodies cross-reactive with myosin.

Anti-group A streptococcal monoclonal antibodies were obtained from BALB c/BYJ mice immunized with purified membranes from M type 5 Streptococcus pyogenes. Two of the anti-streptococcal monoclonal antibodies were previously shown to cross-react with muscle myosin. In this study the monoclonal antibodies were reacted with tissue sections of normal human heart and skeletal muscle. Antibody binding was estimated by indirect immunofluorescence and immunoperoxidase techniques. Both of the monoclonal antibodies (36.2.2 and 54.2.8) investigated in this report reacted with heart and/or skeletal muscle sections. When evaluated by immunofluorescence, monoclonal antibody 54.2.8 demarcated the periphery of cardiac striated muscle cells and reacted to a lesser degree with subsarcolemmal components. Monoclonal antibody 36.2.2 failed to react with heart sections, but both of the monoclonal antibodies reacted strongly with skeletal muscle sections. Results similar to those observed with indirect immunofluorescence were obtained with the immunoperoxidase technique. By Western immunoblotting and competitive inhibition assays, monoclonal antibodies 36.2.2 and 54.2.8 both were found to react with the heavy chain of skeletal muscle myosin. However, only 54.2.8 reacted with the heavy chain of cardiac myosin. The specificity of the monoclonal antibodies for subfragments of skeletal muscle myosin indicated that monoclonal antibody 36.2.2 was specific for light meromyosin fragments, whereas 54.2.8 reacted with both heavy and light meromyosin. The data demonstrated that two monoclonal antibodies against streptococci were specific for skeletal muscle and/or cardiac myosin and for subfragments of the myosin molecule. The reactions of the monoclonal antibodies with human tissue sections were consistent with the immunochemical reactions of the monoclonal antibodies with both denatured and native myosin.