Comparison of ChromID Agar and Clostridium difficile Selective Agar for Effective Isolation of C. difficile from Stool Specimens

BACKGROUND: ChromID Clostridium difficile agar (IDCd; bioMerieux SA, France) is a recently developed chromogenic medium for rapid and specific isolation of C. difficile. We compared the performance of IDCd with that of Clostridium difficile Selective Agar (CDSA). METHODS: A total of 530 fresh stool specimens were collected from patients with clinical signs compatible with C. difficile infection, and cultures for C. difficile were performed on IDCd and CDSA. C. difficile colonies were identified by spore staining, odor, use of an ANI identification test kit (bioMerieux SA), and multiplex PCR for tcdA, tcdB, and tpi. RESULTS: The concordance rate between IDCd and CDSA was 90.6% (480/530). The positivity rates on IDCd on days 1 and 2 (55.6% and 85.0%, respectively) were significantly higher than those on CDSA (19.4% and 75.6%, respectively) (P<0.001 for day 1 and P=0.02 for day 2), but the detection rates on IDCd and CDSA on day 3 were not different (89.4% vs. 82.8%, P=0.0914). On day 3, the recovery rates for non-C. difficile isolates on IDCd and CDSA were 30.2% (160/530) and 22.1% (117/530), respectively (P=0.0075). Clostridium spp. other than C. difficile were the most prevalent non-C. difficile isolates on both media. CONCLUSIONS: The culture positivity rates on IDCd and CDSA were not different on day 3 but IDCd may allow for rapid and sensitive detection of C. difficile within 2 days of cultivation.

Performance of chromID Clostridium difficile Agar Compared with BBL C. difficile Selective Agar for Detection of C. difficile in Stool Specimens

We evaluated the performance of a new chromogenic medium for detection of Clostridium difficile, chromID C. difficile agar (CDIF; bioMerieux, France), by comparison with BBL C. difficile Selective Agar (CDSA; Becton Dickinson and Company, USA). After heat pre-treatment (80degrees C, 5 min), 185 diarrheal stool samples were inoculated onto the two media types and incubated anaerobically for 24 hr and 48 hr for CDIF and for 48 hr and 72 hr for CDSA. All typical colonies on each medium were examined by Gram staining, and the gram-positive rods confirmed to contain the tpi gene by PCR were identified as C. difficile. C. difficile was recovered from 36 samples by using a combination of the two media. The sensitivity with CDIF 48 hr was highest (100%) and was significantly higher than that with CDIF 24 hr (58.3%; P<0.001), because samples with a low burden of C. difficile tended to require prolonged incubation up to 48 hr (P<0.001). The specificity of CDIF 24 hr and CDIF 48 hr (99.3% and 90.6%, respectively) was significantly higher than that of CDSA 48 hr and CDSA 72 hr (72.5% and 67.1%, respectively; P<0.001). CDIF was effective for detecting C. difficile in heat-pretreated stool specimens, thus reducing unnecessary testing for toxin production in non-C. difficile isolates and turnaround time.

Comparison of selective agars recommended by method ISO 11290-1 and chromogenic agars for the isolation of Listeria sp. in refrigerated sausages

The aim of this study was to determine the prevalence of Listeria sp. in refrigerated sausages, and to compare the performance of the selective plating media employed in the ISO 11290-1 method (PALCAM and Oxford agars) with chromogenic agars (Chromogenic Listeria agars CM 1080 (OCLA) and CM 1084). The prevalence of Listeria sp. detected was 52.9%, comprising 13.7% L. monocytogenes strains. The efficacy of the four agars for the isolation of L. monocytogenes proved to be satisfactory. Despite differences in composition of the chromogenic media assessed, these disparities did not affect concordance among results. However, PALCAM agar was shown to suppress other microorganisms more effectively, being more applicable for detecting Listeria strains present in lower quantities. Based on these results, the use of PALCAM agar, in combination with a chromogenic media, is recommended for enhanced isolation of atypical Listeria sp. strains in meat products.

Este estudo teve como objetivo a análise da prevalência de Listeria sp. em linguiças resfriadas e a comparação dos meios seletivos utilizados no plaqueamento do método ISO 11290-1 (Ágar PALCAM e Ágar Oxford), e ágares cromogênicos (Ágares Listeria Cromogênico CM 1080 (OCLA) e CM 1084 (ISO)). A frequência de Listeria sp. foi de 52,9%, sendo que destas, 13,7% corresponderam à L. monocytogenes. A eficácia dos quatro ágares para o isolamento de L. monocytogenes demonstrou-se satisfatória. Apesar de haver algumas diferenças nas composições dos meios cromogênicos analisados, estas não pareceram influenciar nas concordâncias entre os resultados expressos. Contudo, o ágar PALCAM mostrou-se mais eficaz na supressão de outros micro-organismos, aumentando, assim, a possibilidade de detecção de espécies de Listeria presentes em número reduzido. Através deste trabalho sugere-se a utilização do ágar PALCAM associado a um meio cromogênico para aumentar a chance de isolamento de cepas atípicas de Listeria sp. em produtos cárneos.

Evaluation of a ChromID C. difficile Agar for the Isolation of Clostridium difficile / 대한임상미생물학회지

BACKGROUND: Clostridium difficile is the main etiologic agent of antibiotic-associated diarrhea and the most common cause of hospital-acquired diarrhea. Recently, the incidence of C. difficile infections (CDI) has increased and new highly virulent C. difficile strains have emerged. Therefore, accurate and rapid diagnosis is needed. We compared the results of using chromID C. difficile (chromID CD, bioMerieux, France) with the conventional C. difficile Selective Agar (CDSA; BD, USA) for the isolation of C. difficile. METHODS: A total of 738 stool specimens of suspected CDI patients at the Severance Hospital from July to August 2011 were inoculated onto CDSA. Among them, 104 stool specimens revealed colonies on CDSA that were then re-inoculated onto chromID CD. The stool samples were stored at -20degrees C until the time of the re-inoculation. Cultured agars were interpreted after 24 hrs and 48 hrs, respectively. Species identification was performed on the basis of colony characteristics on agar plates as well as the ATB 32A system (API System SA, France). RESULTS: The recovery rates of CDSA and chromID CD were 30.1% and 77.5% after 24 hrs, and 77.5% and 98.6% after 48 hrs, respectively. All of the C. difficile isolates were recovered as typical gray/black colonies on chromID CD. CONCLUSION: The performance of chromID CD for the isolation of C. difficile was better than that of conventional CDSA. The chromID CD could provide easy and sensitive detection of C. difficile even after 24hrs of incubation.

Evaluation of the Usefulness of Selective Chromogenic Agar Medium (ChromID VRE) and Multiplex PCR Method for the Detection of Vancomycin-resistant Enterococci / 대한진단검사의학회지

BACKGROUND: Accurate and early detection of vancomycin-resistant enterococci (VRE) is critical for controlling nosocomial infection. In this study, we evaluated the usefulness of a selective chromogenic agar medium and of multiplex PCR for detection of VRE, and both these techniques were compared with the conventional culture method for VRE detection. METHODS: We performed the following 3 methods for detecting VRE infection in stool specimens: the routine culture method, culturing in selective chromogenic agar medium (chromID VRE, bioMerieux, France), and multiplex PCR using the Seeplex(R) VRE ACE Detection kit (Seegene Inc., Korea) with additional PCR for vanC genes. RESULTS: We isolated 109 VRE strains from 100 stool specimens by the routine culture method. In chromID VRE, all the isolates showed purple colonies, including Enterococcus gallinarum and E. raffinosus, which were later identified using the Vitek card. All VRE isolates were identified by the multiplex PCR method; 100 were vanA-positive E. faecium, 8 were vanA- and vanC-1-positive E. gallinarum, and 1 was vanA-positive E. raffinosus. CONCLUSIONS: For VRE surveillance, culturing the isolates in chromID VRE after broth enrichment appears to be an accurate, rapid, and easy method for routine screening test. Multiplex PCR is relatively expensive and needs skilled techniques for detecting VRE, but it can be an auxiliary tool for rapid detection of genotype during a VRE outbreak.

Usefulness of a Chromogenic Selective Agar for the Identification of Bacillus cereus Isolated from Blood Cultures / 대한진단검사의학회지

BACKGROUND: The incidence of Bacillus cereus bacteremia is increasing, but the identification of Bacillus species remains difficult. Brilliance Bacillus cereus agar (BBC agar; Oxoid, UK) is a new CHROMagar medium that allows selective isolation and identification of B. cereus; however, its clinical usefulness is seldom studied. We evaluated the usefulness of BBC agar to identify B. cereus isolates recovered from blood cultures. METHODS: We analyzed a total of 53 blood isolates that showed a Bacillus-like morphology on Gram staining. All isolates were identified by using both the API Coryne (bioMerieux, France) and API 50CH/B (bioMerieux) systems. They were subsequently subcultured on BBC agar, incubated for 24 hr, and then examined for characteristic blue-green colonies. The clinical characteristics of patients whose isolates were identified as B. cereus were assessed. RESULTS: Of the 53 isolates, 18 were identified as B. cereus by API 50CH/B. With the API 50CH/B system used as gold standard, the sensitivity and specificity for the identification of B. cereus were 100% (18/18) and 100% (35/35), respectively, using BBC agar, and 67% (12/18) and 100% (35/35), respectively, using the API Coryne system. Of the 18 patients with B. cereus bacteremia, 15 showed infectious signs, and 3 had more than 2 blood cultures positive for B. cereus on separate days. CONCLUSIONS: Our study shows, for the first time, that BBC agar, with its good agreement and ease of use, is a valuable alternative to the API 50CH/B system for the presumptive identification of B. cereus isolates from blood cultures.

Evaluation of a New Chromogenic agar, CHROMagar Orientation, for Detection and Presumptive Identification of Urinary Tract Pathogens / 대한진단검사의학회지

BACKGROUND: Urine samples should be tested in a rapid and cost-effective way, as they represent the largest volume of specimens cultured in the microbiology laboratory. Some bacteria can be presumptively identified with CHROMagar Orientation (CO) according to the specific colors produced on the colonies. In this study, the usefulness of CO agar was evaluated for urine cultures. METHODS: The urine samples from 980 patients from March through April, 2004 were inoculated on blood agar (BAP), MacConkey agar, and CO agar plates, and we compared the detection rates of potential pathogens and the agreement between presumptive identification directly from the CO agar and the confirmative idntification, which was performed using Vitek systems (bioMerieux). RESULTS: The detection rates of urinary tract pathogens on all three media, conventional BAP, MacConkey agar and CO agar were identical (18.9%). All isolates of Escherichia coli (54) and ente-rococci (40) were correctly identified with CO agar. The overall agreement of presumptive identification was 87.4% (187/199). CONCLUSIONS: Use of the CO agar enabled a rapid presumptive identification of E. coli, and ente-rococci, the most common urinary tract pathogens. The CO agar is cost-effective by saving some of the bacterial identification kits that would be required for the conventional BAP and MacConkey agar method.