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Comparison between E-test and CLSI broth microdilution method for antifungal susceptibility testing of Candida albicans oral isolates
Koga-Ito, Cristiane Yumi;Lyon, Juliana Pereira;Resende, Maria Aparecida de;
Revista do Instituto de Medicina Tropical de S?o Paulo , 2008, DOI: 10.1590/S0036-46652008000100002
Abstract: thirty candida albicans isolated from oral candidosis patients and 30 c. albicans isolated from control individuals were studied. in vitro susceptibility tests were performed for amphotericin b, fluconazole, 5-flucytosine and itraconazole through the clinical and laboratorial standards institute (clsi) reference method and e test system. the results obtained were analyzed and compared. mic values were similar for the strains isolated from oral candidosis patients and control individuals. the agreement rate for the two methods was 66.67% for amphotericin b, 53.33% for fluconazole, 65% for flucytosine and 45% for itraconazole. according to our data, e test method could be an alternative to trial routine susceptibility testing due to its simplicity. however, it can not be considered a substitute for the clsi reference method.
Comparison of teicoplanin disk diffusion and broth microdilution methods against clinical isolates of Staphylococcus aureus and S. epidermidis
Camargo, Carlos Henrique;Mondelli, Alessandro Lia;B?as, Paulo José Fortes Villas;
Brazilian Journal of Microbiology , 2011, DOI: 10.1590/S1517-83822011000400003
Abstract: the clsi m100-s19 document has recommended the disuse of vancomycin disks for staphylococci and informed that studies on the action of teicoplanin in disk-diffusion testing should be performed. we describe the comparison of two methods, disk diffusion and broth microdilution, for determining teicoplanin susceptibility in clinical isolates of staphylococci. overall results showed an aggregation rate of 96.8%; staphylococcus aureus showed total agreement while s. epidermidis showed 93.8% of agreement. according to these local results, disk diffusion can still be employed to teicoplanin susceptibility determination for staphylococci in our institution.
Comparison of disc diffusion, Etest and broth microdilution for testing susceptibility of carbapenem-resistant P. aeruginosa to polymyxins
Inneke M van der Heijden, Anna S Levin, Ewerton H De Pedri, Liang Fung, Flavia Rossi, Gisele Duboc, Antonio A Barone, Silvia F Costa
Annals of Clinical Microbiology and Antimicrobials , 2007, DOI: 10.1186/1476-0711-6-8
Abstract: Susceptibility of 109 carbapenem-resistant P. aeruginosa to polymyxins was tested comparing broth microdilution (reference method), disc diffusion, and Etest using the new interpretative breakpoints of Clinical and Laboratory Standards Institute.Twenty-nine percent of isolates belonged to endemic clone and thus, these strains were excluded of analysis. Among 78 strains evaluated, only one isolate was resistant to polymyxin B by the reference method (MIC: 8.0 μg/mL). Very major and major error rates of 1.2% and 11.5% were detected comparing polymyxin B disc diffusion with the broth microdilution (reference method). Agreement within 1 twofold dilution between Etest and the broth microdilution were 33% for polymyxin B and 79.5% for colistin. One major error and 48.7% minor errors were found comparing polymyxin B Etest with broth microdilution and only 6.4% minor errors with colistin. The concordance between Etest and the broth microdilution (reference method) was respectively 100% for colistin and 90% for polymyxin B.Resistance to polymyxins seems to be rare among hospital carbapenem-resistant P. aeruginosa isolates over a six-year period. Our results showed, using the new CLSI criteria, that the disc diffusion susceptibility does not report major errors (false-resistant results) for colistin. On the other hand, showed a high frequency of minor errors and 1 very major error for polymyxin B. Etest presented better results for colistin than polymyxin B. Until these results are reproduced with a large number of polymyxins-resistant P. aeruginosa isolates, susceptibility to polymyxins should be confirmed by a reference method.Polymyxins are a multicomponent polypeptide antibiotic that act primarily on the gram-negative bacterial cell wall, leading to rapid permeability changes in the cytoplasmic membrane and ultimately to cell death [1]. The polymyxin E named colistin and polymyxin B have been used to treat several infections caused by multidrug resistant Pseudomonas aerug
Dermatophyte susceptibilities to antifungal azole agents tested in vitro by broth macro and microdilution methods
Siqueira, Emerson Roberto;Ferreira, Joseane Cristina;Pedroso, Reginaldo dos Santos;Lavrador, Marco Aurélio Sicchiroli;Candido, Regina Celia;
Revista do Instituto de Medicina Tropical de S?o Paulo , 2008, DOI: 10.1590/S0036-46652008000100001
Abstract: the in vitro susceptibility of dermatophytes to the azole antifungals itraconazole, fluconazole and ketoconazole was evaluated by broth macro and microdilution methods, according to recommendations of the clsi, with some adaptations. twenty nail and skin clinical isolates, four of trichophyton mentagrophytes and 16 of t. rubrum were selected for the tests. itraconazole minimal inhibitory concentrations (mic) varied from < 0.03 to 0.25 μg/ml in the macrodilution and from < 0.03 to 0.5 μg/ml in the microdilution methods; for fluconazole, mics were in the ranges of 0.5 to 64 μg/ml and 0.125 to 16 μg/ml by the macro and microdilution methods, respectively, and from < 0.03 to 0.5 μg/ml by both methods for ketoconazole. levels of agreement between the two methods (± one dilution) were 70% for itraconazole, 45% for fluconazole and 85% for ketoconazole. it is concluded that the strains selected were inhibited by relatively low concentrations of the antifungals tested and that the two methodologies are in good agreement especially for itraconazole and ketoconazole.
Susceptibility of Dermatophytes to Thiabendazole Using CLSI Broth Macrodilution
Efrén Robledo-Leal,Mariana Elizondo-Zertuche,Gloria M. González
ISRN Dermatology , 2012, DOI: 10.5402/2012/351842
Abstract:
Standartization of broth microdilution method for Mycobacterium tuberculosis
Leite, Clarice Queico Fujimura;Beretta, Ana Laura Remédio Zeni;Anno, Ivone Shizuko;Telles, Maria Alice da Silva;
Memórias do Instituto Oswaldo Cruz , 2000, DOI: 10.1590/S0074-02762000000100021
Abstract: indirect drug susceptibility tests of mycobacterium tuberculosis was done to investigate the accuracy and feasibility of a broth microdilution method (bmm) for determining minimal inhibitory concentrations of conventional drugs against m. tuberculosis. test drugs included isoniazid (h), rifampicin (r), ethambutol (e), streptomycin (s) and pyrazinamide (z). fifty isolates of m. tuberculosis from patients who had never received drug therapy, and h37rv strain for control, were evaluated in the system. when comparing this method with the gold standard proportional method in lowenstein-jensen medium, sensitivity of 100% for all drugs and specifities of 91, 100, 96, 98 and 85% were observed respectively for h, r, e, s and z. the bmm was read faster (14-20 days) than the proportional method (20-28 days). the microdilution method evaluated allows the testing of multiple drugs in multiple concentrations. it is easy to perform and does not require special equipment or expensive supplies. in contrast to radiometric method it does not use radioactive material.
Standartization of broth microdilution method for Mycobacterium tuberculosis
Leite Clarice Queico Fujimura,Beretta Ana Laura Remédio Zeni,Anno Ivone Shizuko,Telles Maria Alice da Silva
Memórias do Instituto Oswaldo Cruz , 2000,
Abstract: Indirect drug susceptibility tests of Mycobacterium tuberculosis was done to investigate the accuracy and feasibility of a broth microdilution method (BMM) for determining minimal inhibitory concentrations of conventional drugs against M. tuberculosis. Test drugs included isoniazid (H), rifampicin (R), ethambutol (E), streptomycin (S) and pyrazinamide (Z). Fifty isolates of M. tuberculosis from patients who had never received drug therapy, and H37Rv strain for control, were evaluated in the system. When comparing this method with the gold standard proportional method in Lowenstein-Jensen medium, sensitivity of 100% for all drugs and specifities of 91, 100, 96, 98 and 85% were observed respectively for H, R, E, S and Z. The BMM was read faster (14-20 days) than the proportional method (20-28 days). The microdilution method evaluated allows the testing of multiple drugs in multiple concentrations. It is easy to perform and does not require special equipment or expensive supplies. In contrast to radiometric method it does not use radioactive material.
Comparison of microdilution broth and disk diffusion for antifungal susceptibility testing against Candida spp.<:Comparison of microdilution broth and disk diffusion for antifungal susceptibility testing against Candida spp. Compara o entre microdilui o e disco difus o para o teste de susceptibilidade aos antifúngicos contra Candida spp.  [cached]
Ant?nio Alexandre de Vasconcelos Júnior,Everardo Albuquerque Menezes,Francisco Afranio Cunha,Maria da Concei??o dos Santos Oliveira Cunha
Semina : Ciências Biológicas e da Saúde , 2012,
Abstract: The evolution of fungal resistance can be evaluated by testing susceptibility to antifungal agents. The protocols for susceptibility testing most common are Broth Microdilution (BMD), Disk Diffusion (DD) and E-test. The objective of this study was to compare the susceptibility tests conducted by the BMD and DD methods for amphotericin B and fluconazole against Candida spp. isolated in Fortaleza/CE. Were used 40 C. albicans, 50 C. tropicalis and 47 C. parapsilosis isolated from blood and urine samples of patients treated at Fortaleza General Hospital. The tests were conducted by the BMD and DD methods according to protocols M27-A3 and M44-A2 of the CLSI. These methods were compared and the percentages of agreement were calculated. For amphotericin B and fluconazole was a predominance of strains with reduced susceptibility. The percentage of agreement for the methods tested was above 97%. No serious errors were detected. The use of DD for performing antifungal susceptibility testing can be applied in routine laboratory tests, since one technique is easy, inexpensive, and reliable when compared to the BMD without compromising the results for the strains of C. albicans, C. tropicalis and C. parapsilosis. A evolu o da resistência fúngica pode ser avaliada pelo teste de susceptibilidade aos antifúngicos. Os protocolos mais utilizados para testes de susceptibilidade s o a micro dilui o em caldo (MDC), disco difus o (DD) e E-test. O objetivo principal deste estudo foi comparar os testes de sensibilidade realizados pelos métodos de MDC e DD para a anfotericina B e o fluconazol contra cepas de Candida spp. isoladas em Fortaleza/CE. Foram utilizadas 40 C. albicans, 50 C. tropicalis e 47 C. parapsilosis, isoladas de amostras de sangue e urina de pacientes atendidos no Hospital Geral de Fortaleza. Os testes foram realizados pelos métodos MDC e DD, de acordo com os protocolos M27-A3 e M44-A2 do CLSI. As metodologias foram comparadas e calculadas os percentuais de concordancia. Para a anfotericina B e fluconazol, ocorreu um predomínio de cepas com susceptibilidade. O percentual de concordancia para as metodologias foi acima de 97% e n o foram detectados erros graves. A utiliza o de DD para realiza o de testes de susceptibilidade antifúngica pode ser aplicada na rotina laboratorial, pois é uma técnica fácil, barata, e de confian a quando comparada ao teste de MDC, sem comprometer os resultados para as cepas de C. albicans, C. tropicalis e C. parapsilosis.
Impact of the New Clinical Breaking Points Proposed by the Clinical and Laboratory Standards Institute (CLSI) in the Antifungal Susceptibility Profile of Clinical Strains Isolated from Invasive Bloodstream Candida Spp. Infections in Spain
R. Cisterna, O. Tellería, S. Hernaez and G. Ezpeleta
Clinical Medicine Insights: Therapeutics , 2012, DOI: 10.4137/CMT.S8293
Abstract: We tested a global collection of Candida spp. strains against fluconazole, voriconazole, caspofungin and anidulafungin using CLSI M27-A3 broth microdilution (BMD) method, in order to to compare if there are any differences between the susceptibility data using the old and new CLSI clinical breakpoints for azoles (fluconazole and voriconazole) and echinocandins (caspofungin and anidulafungin) in a large number of Candida albicans and the non-albicans Candida species emerging in Europe (C. parapsilosis and C. tropicalis) strains isolated from BSIs. During the study period, a total of 919 isolates of Candida spp. (427 C. albicans, 304 C. parapsilosis, 82 C. tropicalis and 106 isolates of other species) were obtained from over 40 Spanish hospitals. The MICs90 (in mg/L) for fluconazole, voriconazole, caspofungin and anidulafungin, respectively, for each considered species were as follows: C. albicans, 2, 0.06, 0.125, 0.125; C. parapsilosis, 2, 0.06, 2, 2; C. tropicalis, 2, 0.125, 0. 125, 0.125. Considering the effects in antifungal susceptibility of the new clinical breakpoints it seems that the new clinical breakpoints are more sensitive in the detection of cross – resistance between different azoles, but contrary to what happened on the susceptibility profile of azoles, the increase in the new clinical breakpoints MIC values decreased the rate of C. parapsilosis echinocandin resistant isolates. Therefore, the new epidemiological clinical breakpoints provided by the CLSI promises to be a more sensitive tool to detect emerging reduced antifungal susceptibility among Candida spp. as well as improve the clinical utility of antifungal in vitro testing.
Investigation of susceptibility of Staphylococcus species to some antibacterial drugs by disk diffusion and broth microdilution  [PDF]
A?anin Jelena,Aksentijevi? Ksenija,?uti? Milenko,Kati? Vera
Veterinarski Glasnik , 2012, DOI: 10.2298/vetgl1204199a
Abstract: The objective of this work was to identify isolated Staphylococcus species and to investigate their sensitivity to some antibacterial drugs. The material used for these investigations were Staphylococcus isolates originating from milk samples. A total of 25 strains of Staphylococcus isolates were examined, including 24 from milk samples from cows with mastitis, and one strain was isolated from a milk sample from a cow following treatment for mastitis. For primary identification, catalase and oxidase tests were used, as well as the free coagulase test. Following the preliminary tests, the isolated strains were identified using commercial systems ID32 STAPH (bioMérieux, France) and the BBL Crystal Gram-Positive ID Kit (Becton Dickinson, USA) according to the enclosed instructions. The Staphylococcus isolates were examined for sensitivity to the following: oxacillin, penicillin, cefoxitin, gentamicin, erythromycin, chloramphenicol, tetracycline, ciprofloxacin, sulfametoxazol/trimetoprim, and vacomycin using the disk diffusion method and the broth microdilution method as recommended by the Clinical and Laboratory Strandards Institute - CLSI(2003), and the results were interpreted according to CLSI recommendations from 2008 and 2010. Antibiogram disks manufactured by Becton Dickinson (USA) were used, and the broth microdilution method was applied using pure antibiotic substances from different manufacturers: erythromycin, chloramphenicol, cefoxitin, gentamicin, oxacillin, tetracycline (Sigma Aldrich, USA), sulfametoxazol (Fluka, USA), penicillin (Calbiochem, Germany), vancomycin (Abbott laboratories, USA), ciprofloxacin and trimetoprim (Zdravlje A.D., Serbia). All 25 strains were catalase positive and oxidase negative. Of the 25 strains, 19 were coagulase positive and 6 were coagulase negative.With the implementation of the disk diffusion method on 19 strains of S. aureus, 17 were established to be resistant to penicillin (89.5%), and 2 strains to gentamicin (10.5%). The investigation of 3 strains of S. xylosus using the disk diffusion method showed that one strain was resistant to tetracycline (33.3%) and to oxacillin (33.3%), while another strain was found to be resistant to penicillin (33.3%). The third strain of S. xylosus was sensitive to all the examined antibiotics. Two strains of S. simulans and one strain of S. haemolyticus were not found to be resistant to any of the examined antibiotics using the disk diffusion method. The implementation of the broth microdilution method yielded in 13 strains of S. aureus resistance to penicillin (68.4%) with MIC
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