ß-lactamase-producing Gram-negative bacteria in an intensive care unit in southern Brazil

ABSTRACT The present study evaluated the antimicrobial susceptibility profile, ß-lactamase production, and genetic diversity of Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter spp. using phenotypic identification, antimicrobial susceptibility testing, and ß-lactamase phenotypic detection. Isolates were obtained from patients in an intensive care unit in a hospital in southern Brazil. Bacterial genomic DNA was extracted, followed by the genotypic detection of carbapenemases and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). Fifty-six isolates (26 Klebsiella pneumoniae, five Escherichia coli, three Enterobacter aerogenes, nine P. aeruginosa, and 13 Acinetobacter spp.) were evaluated. The phenotypic extended spectrum ß-lactamase (ESBL) test was positive in 53.8% of the K. pneumoniae isolates, 100.0% of the E. coli isolates, and 100.0% of the E. aerogenes isolates. Phenotypic and genotypic testing of K. pneumoniae carbapenemase (KPC) was positive in 50.0% of the K. pneumoniae isolates. Phenotypic and genotypic testing showed that none of the P. aeruginosa or Acinetobacter spp. isolates were positive for metallo- ß-lactamase (MBL). The bla OXA gene was detected only in Acinetobacter spp. The lowest genetic diversity, determined by ERIC-PCR, was observed among the KPC-producing K. pneumoniae isolates and OXA-producing Acinetobacter spp. isolates, indicating the inadequate dissemination control of multidrug-resistant bacteria in this hospital environment.

Evaluation of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry-Based VITEK MS System for the Identification of Acinetobacter Species from Blood Cultures: Comparison with VITEK 2 and MicroScan Systems

BACKGROUND: Acinetobacter species are the leading cause of bloodstream infection (BSI), but their correct identification is challenging. We evaluated the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based VITEK MS (bioMerieux, France), and two automated systems, VITEK 2 (bioMerieux) and MicroScan (Siemens, USA) for identification of Acinetobacter BSI isolates. METHODS: A total of 187 BSI isolates recovered at a university hospital in Korea between 2010 and 2012 were analyzed. The identification results obtained using VITEK MS and two automated systems were compared with those of rpoB sequencing. RESULTS: Of 187 isolates analyzed, 176 were identified to the species level by rpoB sequencing: the Acinetobacter baumannii group (ABG; 101 A. baumannii, 43 A. nosocomialis, 10 A. pittii isolates) was most commonly identified (82.4%), followed by Acinetobacter genomic species 13BJ/14TU (5.3%), A. ursingii (2.1%), A. soli (2.1%), A. bereziniae (1.1%), and A. junii (1.1%). Correct identification rates to the species group (ABG) level or the species level was comparable among the three systems (VITEK MS, 90.3%; VITEK 2, 89.2%; MicroScan, 86.9%). However, VITEK MS generated fewer misidentifications (0.6%) than VITEK 2 (10.8%) and MicroScan (13.1%) (P<0.001). In addition, VITEK MS demonstrated higher specificity (100%) for discrimination between ABG and non-ABG isolates than the other systems (both, 31.8%) (P<0.001). CONCLUSIONS: The VITEK MS system is superior to the VITEK 2 and MicroScan systems for identification of Acinetobacter BSI isolates, with fewer misidentifications and better discrimination between the ABG and non-ABG isolates.

Evaluation of VITEK Mass Spectrometry (MS), a Matrix-Assisted Laser Desorption Ionization Time-of-Flight MS System for Identification of Anaerobic Bacteria

BACKGROUND: By conventional methods, the identification of anaerobic bacteria is more time consuming and requires more expertise than the identification of aerobic bacteria. Although the matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems are relatively less studied, they have been reported to be a promising method for the identification of anaerobes. We evaluated the performance of the VITEK MS in vitro diagnostic (IVD; 1.1 database; bioMerieux, France) in the identification of anaerobes. METHODS: We used 274 anaerobic bacteria isolated from various clinical specimens. The results for the identification of the bacteria by VITEK MS were compared to those obtained by phenotypic methods and 16S rRNA gene sequencing. RESULTS: Among the 249 isolates included in the IVD database, the VITEK MS correctly identified 209 (83.9%) isolates to the species level and an additional 18 (7.2%) at the genus level. In particular, the VITEK MS correctly identified clinically relevant and frequently isolated anaerobic bacteria to the species level. The remaining 22 isolates (8.8%) were either not identified or misidentified. The VITEK MS could not identify the 25 isolates absent from the IVD database to the species level. CONCLUSIONS: The VITEK MS showed reliable identifications for clinically relevant anaerobic bacteria.

Identificación de Neisseria gonorrhoeae. Utilidad de la tarjeta 2 NH del sistema Vitek 2C / [Neisseria gonorrhoeae identification. Usefulness of the Vitek 2C NH card].

A total of 115 unique clinical isolates of Neisseria gonorrhoeae and 54 strains of other genera and species included in the database of the NH card were tested by the Vitek 2C System (bioMÞrieux, Marcy L’Etoile, Francia). The gonoccocal isolates had been previously identified by conventional biochemical tests and by the latex agglutination test with monoclonal antibodies using the Phadebact Monoclonal GC Test (Bactus AB, Sweden). The NH card correctly identified 111 (96.5

) strains of 115 isolates; one strain was identified with low discriminatory power (0.86

) was misidentified (as Neisseria meningitidis) whereas the other two (1.7

) remained unidentified. The NH card for N. gonorrhoeae identification provided 100

specificity. The results were available within 6 hours. The NH card could be considered a reliable and useful tool for routine use in Neisseria gonorrhoeae identification.

Evaluation of the BD Phoenix Automated Microbiology System SMIC/ID-2 Panel for Antimicrobial Susceptibility Testing of Streptococcus pneumoniae / 대한진단검사의학회지

BACKGROUND: With the emergence of antimicrobial resistance among Streptococcus pneumoniae, a more accurate and automated antimicrobial susceptibility testing method is essential. We evaluated the BD Phoenix Automated Microbiology System (Becton Dickinson Diagnostic Systems, USA) SMIC/ID-2 panel for antimicrobial susceptibility testing of S. pneumoniae. METHODS: A total of 113 clinical strains of S. pneumoniae (88 penicillin susceptible strains, 8 intermediate strains, and 17 resistant strains by 2008 CLSI criteria) were tested. Minimum inhibitory concentrations (MICs) for penicillin, cefotaxime, clindamycin, erythromycin, levofloxacin, trimethoprim/ sulfamethoxazole, tetracycline, and vancomycin were determined by Etest (AB Biodisk, Sweden) and Phoenix System. The results obtained by Phoenix system were compared to those obtained by Etest. RESULTS: The overall essential agreement of MICs (within one dilution of MICs) defined by the Phoenix and Etest was 92.3%. Neither very major errors nor major errors were produced, and minor errors were 6.5%. Minor errors were frequently observed in susceptibility testings for penicillin (22.1%), cefotaxime (12.4%), and trimethoprim/sulfamethoxazole (11.5%). CONCLUSIONS: The Phoenix SMIC/ID-2 panel provided a simple and rapid susceptibility testing for S. pneumoniae, and the results were in a good agreement with those of Etest. The Phoenix system appears to be an effective automated system in clinical microbiology laboratories.

Evaluation of MicroScan and Phoenix System for Rapid Identification and Susceptibility Testing Using Direct Inoculation from Positive BACTEC Blood Culture Bottles / 대한진단검사의학회지

BACKGROUND: Procedures for rapid identification and susceptibility testing by direct inoculation (DI) from positive blood culture bottles into an automated system have not been standardized. This study was purposed to evaluate DI from BACTEC 9240 blood culture system (BD, USA) into MicroScan (Dade Behring, USA) or Phoenix (BD, USA). METHODS: From May to June 2006, bacterial pellets from positive aerobic bottles showing gram-positive cocci (GPC) or gram-negative rods (GNR) of single morphology were directly inoculated to MicroScan PosCombo1A and NegCombo32 and to Phoenix PMIC/ID-107 and NMIC/ID-53. In addition, the automated instruments were also inoculated from subcultures (standard inoculations, SI). Species identification and susceptibilities were compared between DI and SI and between MicroScan and Phoenix. RESULTS: A total of 108, 104, and 78 specimens were tested with MicroScan, Phoenix, and both, respectively. When DI and SI were matched, 94.8% of GPC were correctly identified with MicroScan, compared to 80.7% with Phoenix, and 93.9% of GNR were correctly identified with MicroScan, compared to 95.7% with Phoenix. DI with MicroScan and Phoenix showed correct susceptibilities in 94.6% of 1,150 and 96.5% of 660 tests (with very major error [VME] of 1.1% and 1.1%), respectively, among GPC and in 94.4% of 942 and 96.3% of 781 tests (with VME of 0.6% and 0%), respectively, of GNR. Correlation of identification/susceptibilities between MicroScan and Phoenix using DI were 81.8%/98.0% for Staphylococcus aureus and 100.0%/95.6% for Escherichia coli. CONCLUSIONS: DI warrants a reliable method for identification and susceptibility testing of both GPC and GNR in MicroScan, and those of only GNR in Phoenix.

Caracterización y resistencia de las cepas de H. influenzae y H. parainfluenzae aisladas de la nasofaringe de portadores / Characterization and resistance of the H. influenzae and H. parainfluenzae isolated from carriers

Introducción: H. influenzae (Hi) y H. parainfluenzae (Hp) forma parte de la flora normal de las vías respiratorias del hombre, el porcentaje de portadores de Hi es muy variable (10-60 por ciento) y depende de la metodología empleada, del grupo etario y del tamaño del universo estudiado. Los niños menores de cinco años están colonizados por esta especie, entre el 10 y 50 por ciento. El serotipo b se asocia a diferentes síndromes y en nuestro país causa del 20 al 45 por ciento de las meningeoencefalitis bacterianas y en las infecciones respiratorias producen morbimortalidad infantil elevada. H. parinfluenzae se ha encontrado asociada a meningitis, artritis, epiglotitis y neumonía, esta bacteria se puede confundir con Hi, la exactitud de estos padecimientos se desconoce. Objetivo. Carat H. influenzae y H. parainfluenzae, aisladas de niños menores de cinco años, mediante pruebas bioquímicas, serológicas, susceptibilidad a los antimicrobianos y producción de beta lactamasa. Material y métodos. Se tomaron 770 exudados nasofaríngeos de niños menores de cinco años y se sembraron en gelosa chocolate bacitracina. La diferenciación de biotipos se realizó por pruebas bioquímicas. Para la tipificación serológica se usó el método de coaglutinación en placa (Phadebact). La beta lactamasa se hizo por tres métodos y la susceptibilidad a los antibióticos por el método de difunsión en disco (Kirby-Bauer modificado). Resultado y discusión. La frecuencia de portadores de H. influenzae fue de 23.7 por ciento y de H. influenzae 16.5 por ciento. En las 159 cepas aisladas, 93 H.influenzae y 66 H parainfluenzae, los biotipos más frecuentes en Hi fueron el V (22.26 por ciento) y el II (20.4 por ciento); los serotipos; Hi tipo b: 30.1 por ciento, del tipo a: 8.6 por ciento, los tipos c-f: 18.2 por ciento y no tipificables: 43.0 por ciento; la beta lactamasa la produjeron Hi en el 27.0 por ciento y Hp en el 18.0 por ciento. Guiscafré y col, reportaron 14.0 por ciento de resistencia a la penicilina en cepas aislada de niños, en el presente encontramos 16.0 por ciento; al clorafenicol sólo el 5.0 por ciento de Hi fue resistente, sin embargo Campos y col. informaron de un 52.0 por ciento en España. Sensibles a la ticarcilina/ac. clavulánico, el 100.0 por ciento; a las cefalosporinas de segunda y tercera generación el 97.0 por ciento y 95.0 por ciento respectivamente

Evaluación del uso de un esquema de pruebas bioquímicas para tipificar estreptococos del grupo "viridans" y Streptococcus bovis / Use of a set of biochemical test for typing \"viridans\" group streptococci and Streptococcus bovis

Con el fin de evaluar un esquema de pruebas bioquímicas para identificar estreptococos del grupo "viridans" y S. bovis, se tipificaron 110 cepas aisladas de materiales clínicos. El 74,5 por ciento fue identificado con criterio de coincidencia absoluta con el esquema utilizado, el 17,3 por ciento se tipificó admitiendo 1 reacción atípica y el 8,2 por ciento fue nominado como estreptococos, puesto que no pudieron ser incluidos en ninguno de los dos casos anteriores. Las especies que más frecuentemente mostraron una reacción atípica, fueron S. mitis (7/22 cepas), S. anginosus (S. milleri) (6/16 cepas) y S. sanguis (3/16 cepas). Del total de cepas de S. sanguis, S. mutans y S. bovis I, aisladas se hemocultivo, el 86 por ciento estuvo asociado a Endocarditis Infecciosa. La capacidad para producir dextrán en las cepas aisladas de hemocultivo, se asoció a la presencia de esta patología, con valores predictivos positivo y negativo de 86 por ciento y 94 por ciento, respectivamente. S. anginosus (S. milleri) se aisló de abscesos más frecuentemente que las otras especies. Consideramos que debido a la transcendencia clínica que tiene actualmente la correcta identificación a nivel de especie de estos microorganismos, el esquema propuesto constituye una alternativa útil para tal fin, en los laboratorios de Bacteriología Clínica de nuestro medio