[A simple disk diffusion test to identify beta-lactamase-negative, ampicillin-resistant Haemophilus influenzae--application of cephalexin, cefsulodin and cefaclor disks].

Currently, beta-lactamase-negative (BLN) ampicillin-resistant (AR) strains of Haemophilus influenzae are prevalent in Japan. BLNAR strains are defined by the presence of specific mutation(s) in the ftsI gene but are not phenotypically distinguishable by ampicillin (ABPC) susceptibility. In the present study, we showed that cephalexin (CEX), cefsulodin (CFS), and cefaclor (CCL) disk diffusion tests can be effectively used to identify BLNAR strains. A total of 169 clinical isolates of BLN H. influenzae, consisting of 113 of BLNAR and 56 of BLN, ampicillin-susceptible (AS), were included. All the isolates were genetically defined by detection of the TEM gene and partial sequencing of the ftsI gene. The Clinical and Laboratory Standards Institute (CLSI) standard broth microdilution and disk diffusion tests for ABPC provided 20% and 19% false susceptible rates, respectively. Alternatively, 34 cephem agents were tested using disk diffusion. Of the agents tested, CEX, CFS, and CCL disks could effectively discriminate between BLNAR and BLNAS isolates. All the BLNAS isolates showed visible growth inhibitory zones around CEX and CFS disks, but 108 (95.6%) and 106 (93.8%) BLNAR isolates did not. The results indicated 100% predictive values (PVs) for BLNAR and PVs for BLNAS were 91.8% for CEX and 88.9% for CFS. The CLSI-based interpretations for CCL (> or =20 mm) also highly correlated with BLNAR and BLNAS, PVs for BLNAR and for BLNAS being 100% and 93.3%, respectively. With simplicity and discriminability of the test method, we recommend a CEX disk diffusion test in combination with a rapid beta-lactamase test to identify BLNAR isolates in clinical laboratories.

[Laboratory-based evaluation of significance to routinely use anaerobic blood culture bottles: analysis of positivity and rapidity to detect positive cultures].

The publications in 1990s have indicated decreased recovery rates of obligate anaerobes from blood cultures and have questioned the need for routine anaerobic blood culture bottles. In this study, we compared positivities of the paired aerobic and anaerobic bottles and rapidity to detect positive cultures by two automated blood culture systems, BACTEC 9120 and BacT/ALERT 3D. Of 401 positive readings by BACTEC 9120, 338(84.3%) aerobic bottles became to be positive, and anaerobic bottles were 318(79.3%). Also, of 437 positive readings by BacT/ALERT 3D, positivities were 90.8% and 67.3% by aerobic and anaerobic bottles, respectively. These results indicated 5.0% and 23.7% more organisms were recovered in aerobic bottles than in anaerobic bottles, including more staphylococci, gram-positive rods, glucose-nonfermentative gram-negative rods and yeasts. Only 4 (0.14%) of 2,799 BACTEC 9120 anaerobic bottles and 2 (0.06%) of 3,428 BacT/ALERT 3D anaerobic bottles recovered obligate anaerobes. We compared time to detect positive cultures during incubation cycle by both aerobic and anaerobic bottles. Aerobic bottles in BACTEC 9120 read more positive cultures >2 hours earlier than anaerobic bottles, whereas BacT/ALERT 3D could not demonstrate a statistical significance in rapid reading of positive cultures. These results support that recovery rates of obligate anaerobes markedly decreased and that the routine use of anaerobic blood culture bottles is not legitimate at this time. In place of anaerobes, it is an urgent and important issue how to recover fungi correctly and rapidly from blood cultures.