bWe analyzed the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of smudge plate growth for bacterial identification from 400 blood cultures. Ninety-seven percent of Gram-negative bacilli and 85% of Gram-positive organisms were correctly identified within 4 h; only eight isolates (2.0%) were misidentified. This method provided rapid and accurate microbial identification from positive blood cultures. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is used in clinical microbiology laboratories for rapid identification of microbial isolates grown in culture (1, 2). Implementation of MALDI-TOF MS in the microbiology laboratory in conjunction with antibiotic stewardship has been associated with earlier initiation of effective antimicrobial therapy and lower 30-day mortality in patients with bloodstream infections (3). This technology has also been applied to reduce turnaround times for identification of blood culture isolates directly from positive blood culture broths (4-16). Various techniques have been assessed, generally involving lysis/centrifugation of blood culture pellets in preparation for analysis by MALDI-TOF MS. The current study evaluated a method using "smudge" plates for subsequent analysis with MALDI-TOF MS to simplify sample processing and to improve the ability to rapidly identify bacteria from positive blood cultures.Blood cultures were collected in Bactec Plus aerobic and anaerobic bottles incubated in the Bactec 9240 system (Becton Dickinson, Franklin Lakes, NJ). We prospectively examined 400 blood cultures that were flagged as positive for bacterial growth between 8:00 a.m. and 3:00 p.m. on weekdays from 1 April to 30 September 2014. A 1-to 2-ml aspirate from the blood culture bottle was used to prepare a Gram stain and was subcultured to blood, chocolate, and MacConkey agar plates. The blood and MacConkey plates were incubated at 35°C in ambient air; chocolate agar plates were incubated at 35°C in 5% CO 2 . Brucella agar plates incubated anaerobically were added for subculture from positive anaerobic blood culture bottles. Aerobic and facultative organisms were identified using standard phenotypic methods, including coagulase, oxidase, latex agglutination, streptococcal serotyping, the Vitek 2 system (bioMérieux, Durham, NC), API strips (bioMérieux), and other biochemical tests as appropriate. Anaerobes were identified using the Remel RapID Ana II (Oxoid, Hampshire, United Kingdom).A smudge plate was prepared when a single morphology was evident on the Gram stain; specimens with more than one bacterial morphology, yeasts, or filamentous fungi were excluded from this study. If two blood culture bottles from the same set were positive, only the aerobic bottle was used for smudge plate preparation, and multiple positive blood cultures obtained within 24 h from the same patient were included in this study only once. For smudge plate preparation, 3 ml of blood culture broth was aspirated from positive blood cultu...
We compared StrepB Select medium (Select) after enrichment with conventional culture for the detection of Group B Streptococcus (GBS). Postenrichment sensitivities of Select and conventional culture were 98.8% and 92.2%, respectively (P < 0.05). Select was superior for detection of GBS from vaginal-rectal specimens. Growth of non-GBS colonies required additional work to exclude the presence of GBS, especially after 48 h of incubation. Incubation of Select beyond 24 h did not significantly increase the yield of GBS.As group B Streptococcus (GBS) remains a significant cause of neonatal morbidity and mortality, antenatal screening for GBS at 35 to 37 weeks of gestation is recommended to determine whether antimicrobial prophylaxis is warranted (4, 7). The use of newer, chromogenic media may improve the yield of GBS, while reducing labor and turnaround time (5). We evaluated a new chromogenic medium, StrepB Select (Select; Bio-Rad Laboratories, Marnes-la-Coquette, France), a selective medium for the detection and presumptive identification of GBS in vaginal and vaginal-rectal specimens (9). We compared the recovery of GBS from StrepB Select with that from conventional culture on colistin-nalidixic acid agar with 5% sheep blood (CNA; Oxoid, Nepean, Ontario, Canada) with and without broth enrichment using Streptococcus selective broth (SSB; Bio-Media Ltd., Woodbridge, Ontario, Canada).From September to November 2008, a total of 1,025 specimens from 992 patients were submitted for GBS screening. These swabs were directly inoculated onto CNA plates and then placed into enrichment broth (SSB). After 24 h of incubation, the direct CNA plate was examined for the presence of colonies suggestive of GBS. If direct culture on CNA did not yield GBS, the SSB was subcultured onto CNA and incubated for 24 h. All broths were subcultured onto StrepB Select and incubated for up to 48 h at 37°C in ambient air, per the manufacturer's recommendations. Colonies suggestive of GBS, with a turquoise blue color on Select and gray colonies with or without hemolysis on CNA, were worked up by separate, experienced technologists blinded to each other's work. Identification of GBS was performed using conventional tests, including those with catalase, Gram stain, and Lancefield grouping antisera by using the PathoDx latex agglutination kit (Remel, Inc., Lenexa, KS). PCR testing for the cfb gene encoding the Christie-Atkins-Munch-Petersen (CAMP) factor (3), directly from the SSB enrichment broth, was used as the gold standard. A true positive was defined as growth of GBS on either medium.Of the 1,025 specimens tested, a total of 243 (23.7%) yielded GBS, and the same 243 specimens were also positive by PCR for the cfb gene. Direct culture onto CNA yielded GBS from 201 samples (82.7%) ( Table 1). SSB enrichment with CNA subculture at 24 h detected an additional 23 isolates (224/243 samples; 92.2%), while SSB enrichment with Select subculture detected GBS in 240 of 243 samples, for a sensitivity of 98.8% (P Ͻ 0.0001). There were no specimens that yield...
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