BACKGROUND: The rapid and accurate detection of carbapenemase-producing Enterobacteriaceae (CPE) in clinical microbiology laboratories is essential for the treatment and control of infections caused by these microorganisms. This study was performed to evaluate the ability of the VITEK AST-N202 card to detect CPE isolates. MATERIALS AND METHODS: A total of 43 (Klebsiella pneumoniae, n = 37; Escherichia coli, n = 3; and Enterobacter cloacae, n = 3) CPE isolates and 79 carbapenemase-non-producing Enterobacteriaceae (CNE) isolates were included in this study. The CPE isolates harbored KPC-2 (n = 11), KPC-3 (n = 20), GES-5 (n = 5), VIM-2 (n = 2), IMP-1 (n = 1), NDM-1 (n = 2), or OXA-232 (n = 2). Of the 79 CNE isolates, eight K. pneumoniae isolates were resistant to ertapenem, imipenem, and meropenem, while the remaining 71 isolates were susceptible to the carbapenems. Antimicrobial susceptibilities were tested using the VITEK AST-N202 card, and the results were interpreted as positive when the isolates showed resistant or intermediate results. Modified-Hodge tests (MHTs) were performed using ertapenem or meropenem disks for the screening of carbapenemase production. Polymerase chain reaction (PCR) and direct sequencing were used to identify beta-lactamase genes. RESULTS: Sensitivity of MHT with ertapenem and meropenem disks for the detection of carbapenemase was 81.4% (35/43) and 81.4% (35/43), respectively, and a combination with both antibiotic disks increased the sensitivity to 88.4% (38/43). Specificity of the MHT was 100% (79/79) for the CNE isolates. Sensitivity of ertapenem, imipenem, and meropenem as assessed by the VITEK AST-N202 card was 100% (43/43), 93% (40/43), and 95.3% (41/43), respectively. Specificity (89.8%, 71/79) of the test with each carbapenem was improved to 100% (71/71) when eight carbapenem-resistant CNE isolates were excluded from the testing. CONCLUSION: The VITEK AST-N202 card showed high sensitivity for the detection of carbapenemases in Enterobacteriaceae strains. PCR and sequencing experiments for the detection of carbapenemases are recommended when clinical Enterobacteriaceae isolates show non-susceptibility to carbapenems.
beta-Lactamases ; Carbapenems ; Enterobacter cloacae ; Enterobacteriaceae* ; Escherichia coli ; Imipenem ; Mass Screening ; Pneumonia ; Polymerase Chain Reaction ; Sensitivity and Specificity

BACKGROUND: Fecal occult blood tests have been widely used to screen for colorectal cancer. SENTiFIT 270 (Sentinel diagnostics, Italy) is a fecal occult blood test with an immunochemical method that utilizes FOB Gold reagents. We evaluated the performance of SENTiFIT 270 using the FOB Gold reagent. In addition, FOB Gold was evaluated with the HITACHI 7180 (Hitachi Ltd., Japan). METHODS: The precision and linearity of the SENTiFIT 270 was evaluated in accordance with applicable Clinical and Laboratory Standard Institute guidelines. The comparison study between SENTiFIT 270-FOB Gold and the OC-Sensor (Eiken chemical Co., Japan) was performed using stool specimens. RESULTS: In the precision evaluation, the total precision of SENTiFIT 270-FOB Gold was 4.94% and 2.54% at high and low concentrations, respectively. The HITACHI 7180-FOB Gold had excellent precision of 4.60% and 2.09% at high and low concentrations, respectively. Linearity was also excellent for the SENTiFIT 270-FOB Gold and HITACHI 7180-FOB Gold at 0.9987 and 0.9986, respectively. The SENTITIF 270-FOB Gold showed excellent agreement with a kappa value of 0.830 and a concordance rate of 93.6%. The HITACHI 7180-FOB Gold showed high agreement with a kappa value of 0.832 and a concordance rate of 93.9%. CONCLUSION: The SENTiFIT 270-FOB Gold showed excellent performance in accuracy, linearity, and comparative inspection ability.
Colonic Neoplasms ; Colorectal Neoplasms ; Indicators and Reagents ; Methods ; Occult Blood

Annual external quality assessment was performed three times for clinical microbiology division of The Korean Association of Quality Assurance for Clinical Laboratory. For each trial, three sets composed of different combinations of four bacteria and one yeast were distributed for culture, identification, and antimicrobial susceptibility tests. A total of 340 laboratories were enrolled and 330 (97.0%), 331(97.4%), and 331(97.4%) returned the results on trial I, II, and III, respectively. For bacterial identification, the correct identification of gram-negative bacilli, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus capitis, Streptococcus agalactiae, Listeria monocytogenes, and Candida species was greater than 95%. However, correct identification of Staphylococcus lugdunensis, Corynebacterium striatum, Vibrio vulnificus, Aeromonas hydrophila, Cryptococcus neoformans, and Malassezia pachydermatis was relatively less accurate, with values of 95.4%, 89.9%, 50.7%, 91.3%, 93.6%, and 93.9%, respectively. Surveillance cultures for vancomycin-resistant enterococci and methicillin-resistant S. aureus were correctly determined by 95.4% and 93.9% of the respondents, respectively. False carbapenem-resistance due to AmpC beta-lactamase, disk diffusion testing for vancomycin in Staphylococcus species, oxacillin and penicillin susceptibility testing in S. lugdunensis and false imipenem-resistance in Proteus species were common sources of inaccurate results. The accuracy of species identification for Corynebacterium species and Vibrio species requires improvement. Consistent problems occurred with antimicrobial susceptibility testing of vancomycin for Staphylococcus species using the disk diffusion method.
Aeromonas hydrophila ; Bacteria ; beta-Lactamases ; Candida ; Corynebacterium ; Cryptococcus neoformans ; Surveys and Questionnaires ; Diffusion ; Korea ; Listeria monocytogenes ; Malassezia ; Methicillin Resistance ; Oxacillin ; Penicillins ; Proteus ; Staphylococcus ; Staphylococcus aureus ; Staphylococcus epidermidis ; Staphylococcus lugdunensis ; Streptococcus agalactiae ; Vancomycin ; Vibrio ; Vibrio vulnificus ; Yeasts

Annual external quality assessment was performed three times for clinical microbiology division of The Korean Association of Quality Assurance for Clinical Laboratory. For each trial, three sets composed of different combinations of four bacteria and one yeast were distributed for culture, identification, and antimicrobial susceptibility tests. A total of 340 laboratories were enrolled and 330 (97.0%), 331(97.4%), and 331(97.4%) returned the results on trial I, II, and III, respectively. For bacterial identification, the correct identification of gram-negative bacilli, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus capitis, Streptococcus agalactiae, Listeria monocytogenes, and Candida species was greater than 95%. However, correct identification of Staphylococcus lugdunensis, Corynebacterium striatum, Vibrio vulnificus, Aeromonas hydrophila, Cryptococcus neoformans, and Malassezia pachydermatis was relatively less accurate, with values of 95.4%, 89.9%, 50.7%, 91.3%, 93.6%, and 93.9%, respectively. Surveillance cultures for vancomycin-resistant enterococci and methicillin-resistant S. aureus were correctly determined by 95.4% and 93.9% of the respondents, respectively. False carbapenem-resistance due to AmpC beta-lactamase, disk diffusion testing for vancomycin in Staphylococcus species, oxacillin and penicillin susceptibility testing in S. lugdunensis and false imipenem-resistance in Proteus species were common sources of inaccurate results. The accuracy of species identification for Corynebacterium species and Vibrio species requires improvement. Consistent problems occurred with antimicrobial susceptibility testing of vancomycin for Staphylococcus species using the disk diffusion method.
Aeromonas hydrophila ; Bacteria ; beta-Lactamases ; Candida ; Corynebacterium ; Cryptococcus neoformans ; Surveys and Questionnaires ; Diffusion ; Korea ; Listeria monocytogenes ; Malassezia ; Methicillin Resistance ; Oxacillin ; Penicillins ; Proteus ; Staphylococcus ; Staphylococcus aureus ; Staphylococcus epidermidis ; Staphylococcus lugdunensis ; Streptococcus agalactiae ; Vancomycin ; Vibrio ; Vibrio vulnificus ; Yeasts

Annual proficiency surveys were conducted in March, June, and September in 2015 by the Clinical Microbiology Subcommittee of the Korean Association of External Quality Assessment Service. The program covers the sections of bacteriology, advanced bacteriology and mycology, mycobacteriology, and parasitology. Each trial was composed of three sets of different combinations of five bacteria and yeasts. These sets were distributed among laboratories for Gram staining, culture, identification, and antimicrobial susceptibility tests. Five slides with fixed sputum smears were provided as part of each trial for acid-fast bacilli detection. The survey material distribution was section-based. Two survey materials were provided in each trial, while five specimens for mycobacterial culture and identification, five specimens for anti-tuberculosis susceptibility testing and two Mycobacterium tuberculosis strains for rapid detection of rifampin and isoniazid resistance were distributed in the March and June trials. Five virtual microscopy files for stool parasite examination were availed by registered participants in the June trial. Out of the 334 enrolled laboratories, 328 (98.2%), 328 (98.2%), and 329 (98.5%) submitted responses in trials I, II, and III, respectively. Identification of bacteria, namely, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Pseudomonas aeruginosa, Streptococcus pneumoniae, and Vibrio fluvialis by more than 95% of participants was acceptable. Surveillance cultures for vancomycin-resistant enterococci and carbapenem-resistant Enterobacteriaceae were determined accurately by 75.8%–85.3% and 93.1% of the respondents, respectively. Species-level identification of Candida krusei, Candida lusitanae, and Candida guilliermondii was still low at 79.8%, 55.7%, and 42.7%, respectively. Disk diffusion method revealed an unacceptably high false-positive rate of resistance to glycopeptides in E. faecalis and to trimethoprim-sulfamethoxazole in S. pneumoniae. Advanced bacteriology trials revealed unsatisfactory results for species-level identification of moulds. Mycobacterial culture, identification and susceptibility testing, and molecular detection of rifampin and isoniazid resistance were performed exceedingly well by participants. Hymenolepsis diminuta could not be identified by participants, with a correct answer rate of only 46.5% and ‘no parasite seen’ answer rate of only 31.8% for negative specimens. Species-level identification of Candida and moulds was challenging for clinical microbiology laboratories. Disk diffusion method was found to be problematic in testing the susceptibility of microorganisms to glycopeptides and trimethoprim-sulfamethoxazole. Improvement is required in result interpretation of negative specimens in parasitology.
Bacteria ; Bacteriology ; Candida ; Diffusion ; Enterobacteriaceae ; Enterococcus faecalis ; Escherichia coli ; Glycopeptides ; Isoniazid ; Klebsiella pneumoniae ; Korea* ; Methods ; Microscopy ; Mycobacterium ; Mycobacterium tuberculosis ; Mycology ; Parasites ; Parasitology ; Pneumonia ; Pseudomonas aeruginosa ; Quality Control ; Rifampin ; Sputum ; Streptococcus pneumoniae ; Surveys and Questionnaires ; Trimethoprim, Sulfamethoxazole Drug Combination ; Vancomycin-Resistant Enterococci ; Vibrio ; Yeasts

Annual proficiency surveys were performed in March, June and September 2014 by clinical microbiology division of The Korean Association of Quality Assurance for Clinical Laboratory. Parasitology part has been newly incorporated in this survey. For each trial, three sets which were composed of different combinations of five bacteria and yeast were distributed for gram stain, culture, identification, and antimicrobial susceptibility tests of general bacteriology and five fixed sputum smear on slides were distributed for acid fast bacilli stain. Two advanced bacteriology survey materials for culture and identification of anaerobic bacteria and mold were distributed to the voluntary participants in every trial and five mycobacterial culture and identification specimens, five anti-tuberculosis susceptibility testing specimens, and two Mycobacterium tuberculosis strains for rapid detection of rifampin and isoniazid resistance were distributed to the voluntary participants in March and June trials. Five virtual microscopic slides for stool parasite examination were open for the registered participants in June trial. A total of 340 laboratories were enrolled and 330 (97.0%), 331 (97.4%), and 331 (97.4%) returned the results on trial I, II, and III, respectively. For bacterial identification, the percent acceptable identification of Burkholderia cepacia, Klebsiella pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pneumoniae, Streptococcus agalactiae, Plesiomonas shigelloides, and Enterococcus faecalis were greater than 95%. Group C and group D Salmonella species challenged as the different sets of M1422 resulted in the acceptable rate lower than 95% because nine participants reported the identification of different sets. Surveillance cultures for methicillin-resistant S. aureus and vancomycin-resistant enterococci were correctly determined by 89.6% and 69.0% of the respondents, respectively. Correct identification to species level of Candida albicans, Candida auris, Candida glabrata, and Candida parapsilosis were 86.1%, 1.6%, 48.1%, and 83.8%. Vancomycin disk diffusion test in S. aureus, missing oxacillin screen or penicillin susceptibility test in S. pneumoniae and lack of reliable methods of quinolone resistance detection in Salmonella species caused unacceptable results in antimicrobial susceptibility testing. Advanced bacteriology trials revealed low performance in species identification of mold. Mycobacterial culture, identification and susceptibility test performance was kept in excellence. The performance of identification of stool parasites was acceptable >90% for detection of helminth eggs and amebic cysts but 28.6% false positive responses resulted from negative specimens. In conclusion, species-level identification of fungi of both candida species and mold were challenging to clinical microbiology laboratories. Vancomycin disk diffusion method for S. aureus and lack of proper penicillin susceptibility test for S. pneumoniae were still common cause of inaccurate results. Virtual microscopic survey has been successfully introduced in parasitology.
Bacteria ; Bacteria, Anaerobic ; Bacteriology ; Burkholderia cepacia ; Candida ; Candida albicans ; Candida glabrata ; Surveys and Questionnaires ; Diffusion ; Eggs ; Enterococcus faecalis ; Fungi ; Helminths ; Isoniazid ; Klebsiella pneumoniae ; Korea* ; Methicillin Resistance ; Mycobacterium tuberculosis ; Ovum ; Oxacillin ; Parasites ; Parasitology ; Penicillins ; Plesiomonas ; Pneumonia ; Pseudomonas aeruginosa ; Rifampin ; Salmonella ; Sputum ; Staphylococcus aureus ; Streptococcus agalactiae ; Streptococcus pneumoniae ; Streptococcus pyogenes ; Vancomycin ; Yeasts