scholarly journals Department of Microbiology, Bangladesh Agricultural University, Mymensingh

2016 ◽  
Vol 2 (2) ◽  
pp. 3-6
Author(s):  
Nahida Akther Zahan ◽  
Md. Akram Hossain ◽  
AKM Shamsuzzaman ◽  
AKM Musa ◽  
Md. Chand Mahamud ◽  
...  
Keyword(s):  
Clinical Laboratory ◽  
Methicillin Resistance ◽  
Meca Gene ◽  
Toxin Production ◽  
Latex Agglutination ◽  
Diffusion Method ◽  
National Committee ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method

The study was done to detect different exotoxins among the strains of Staphylococcus aureus isolated in the department of Microbiology, Mymensingh Medical College in collaboration with the Department of Medicine under the Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh between the periods from July, 2006 to June, 2007. A total of 40 S. aureus isolates investigated in this study were identified by standard microbiological techniques. Antimicrobial susceptibility of the isolates to Oxacillin was carried out by disk diffusion method as per recommendation of the National Committee for Clinical Laboratory Standards. Any isolate showing resistance to Oxacillin was tested again by agar dilution method to determine minimum inhibitory concentration (MIC) of Methicillin. All strains were also tested for mecA gene by Polymerase Chain Reaction (PCR) for confirmation of Methicillin resistance. Enterotoxin (A-D) and Toxic Shock Syndrome Toxin-1 (TSST-1) were detected by Reverse Passive Latex Agglutination (RPLA) test. Out of 40 S. aureus isolates, 7 (17.5%) Methicillin Resistant S. aureus (MRSA), 1 (2.5%) Methicillin Sensitive S. aureus (MSSA) produced Staphylococcal Enterotoxin A (SE-A) and 1 MRSA isolate was positive for TSST-1. In case of combined toxin production among the S. aureus isolates, 2 (5%) MSSA were found to produce SE-A and SE-B, 2 (5%) MSSA produced SE-C and SE-D, and 1 (2.5%) MRSA, 1 (2.5%) MSSA produced SE-C and TSST-1.Bangladesh J Med Microbiol 2008; 02 (02): 3-6

scholarly journals Antimicrobial Susceptibilities and Serogroups of Clinical Strains of Clostridium difficile Isolated in France in 1991 and 1997

1999 ◽  
Vol 43 (11) ◽  
pp. 2607-2611 ◽  
Author(s):  
Frédéric Barbut ◽  
Dominique Decré ◽  
Béatrice Burghoffer ◽  
Danièle Lesage ◽  
Françoise Delisle ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Clinical Laboratory ◽  
Major Change ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
National Committee ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method ◽  
Therapeutic Drugs

ABSTRACT Glycopeptides (vancomycin and teicoplanin) and metronidazole are the drugs of choice for the treatment of Clostridium difficile infections, but trends in susceptibility patterns have not been assessed in the past few years. The objective was to study the MICs of glycopeptides and metronidazole for unrelated C. difficile strains isolated in 1991 (n = 100) and in 1997 (n = 98) by the agar macrodilution, the E-test, and the disk diffusion methods. Strain susceptibilities to erythromycin, clindamycin, tetracycline, rifampin, and chloramphenicol were also determined by the ATB ANA gallery (bioMérieux, La Balme-les-Grottes, France). The MICs at which 50% of isolates are inhibited (MIC50s) and MIC90s of glycopeptides and metronidazole remained stable between 1991 and 1997. All the strains were inhibited by concentrations that did not exceed 2 μg/ml for vancomycin and 1 μg/ml for teicoplanin. Comparison of MICs determined by the agar dilution method recommended by the National Committee for Clinical Laboratory Standards and the E test showed correlations (±2 dilutions) of 86.6, 95.9, and 99% for metronidazole, vancomycin, and teicoplanin, respectively. The E test always underestimated the MICs. Strains with decreased susceptibility to metronidazole (MICs, ≥8 μg/ml) were isolated from six patients (n = 4 in 1991 and n = 2 in 1997). These strains were also detected by the disk diffusion method (zone inhibition diameter, ≤21 mm); they belonged to nontoxigenic serogroup D (n = 5) and toxigenic serogroup H (n = 1). Decreased susceptibility to erythromycin (MICs, ≥1 μg/ml), clindamycin (MICs, ≥2 μg/ml), tetracycline (MICs, ≥8 μg/ml), rifampin (MICs, ≥4 μg/ml), and chloramphenicol (MICs, ≥16 μg/ml) was observed in 64.2, 80.3, 23.7, 22.7, and 14.6% of strains, respectively. Strains isolated in 1997 were more susceptible than those isolated in 1991, and this trend was correlated to a major change in serogroup distribution. Periodic studies are needed in order to detect changes in serogroups and the emergence of strains with decreased susceptibility to therapeutic drugs.

scholarly journals Prevalence, Antibiogram and Molecular Characterization of Comunity-Acquired Methicillin-Resistant Staphylococcus Aureus in AWKA, Anambra Nigeria

2016 ◽  
Vol 10 (1) ◽  
pp. 211-221 ◽  
Author(s):  
Blessing Ike ◽  
Malachy C. Ugwu ◽  
Moses N. Ikegbunam ◽  
David Nwobodo ◽  
Chika Ejikeugwu ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Binding Protein ◽  
Meca Gene ◽  
Latex Agglutination ◽  
Diffusion Method ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Penicillin Binding Protein ◽  
Carriage Rate ◽  
Molecular Features

Objectives:This study evaluated the prevalence, antibiogram and molecular features of CA-MRSA in Awka, Nigeria.Methods:Confirmation of MRSA was done by testing resistance to oxacillin (1µg), cloxacillin (5µg) and cefoxitin(30µg) on sterile Mueller Hinton agar supplemented with 4% sodium chloride. The MRSA strains were subjected to antimicrobial susceptibility testing using Kirby-Bauer disc diffusion method. Minimum inhibitory concentration was determined using agar dilution method. Penicillin binding protein 2a was detected through rapid latex agglutination assay while mecA gene was detected by polymerase chain reaction. A total of 142S. aureusisolates were obtained from 261 samples sourced from Staff, students and fomites of the Faculty of Pharmaceutical SciencesResult:The overall prevalence of MRSA was 22.6%. The carriage rate was higher in females (56.5%) than male (43.5%) and was highest in individuals of 20-30 years of age (57.65%). The MIC of the oxacillin sodium salt ranged from 4-32 μg/ml. The multi-antibiotic resistance indices show that 53.4% had Multiple Antibiotic Resistance Indexing (MARI) higher than 0.2. Penicillin binding protein 2a was detected in 8.4% of MRSA isolates, all from nasal carriage while mecA gene was detected in 5 of isolates.Conclusion:This study showed a very high prevalence of MRSA carriage among studied subjects.

scholarly journals Comparison of rapid and conventional methods for investigating of mecA presence in Staphylococcus Species

2021 ◽  
Vol 37 (5) ◽  
Author(s):  
Selim Görgün ◽  
Hacer İşler ◽  
Mehmet Cenk Turgut
Keyword(s):  
Methicillin Resistance ◽  
Meca Gene ◽  
Latex Agglutination ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
Chromogenic Agar ◽  
Pcr Method ◽  
Conventional Methods

Objectives: Taking the determination of mecA gene by polymerized chain reaction (PCR) method as a reference in determining methicillin resistance in Staphylococcus species, we aimed at comparing the reliability levels of disk diffusion, latex agglutination test and chromogenic agar use methods. Methods: This prospective study was conducted on 228 Staphylococcus strains isolated between January 2020 and December 2020 in Samsun Training and Research Hospital. Disk diffusion, latex agglutination and chromogen agar medium methods were applied along with the polymerized chain reaction (PCR) method. Results: The mecA gene was detected in 47 of the isolates (20.6%) by the PCR method, and these isolates were accepted as methicillin-resistant. When the PCR result was taken as a reference, the sensitivity of the disk diffusion method became 100%, and specificity became 45.9%; sensitivity of latex agglutination was determined as 80.9%, and specificity as 70.2%; sensitivity of chromogenic agar as 85.1% and its specificity was found to be 95%. Only in S. aureus isolates, the highest sensitivity and specificity rate (100% and 88%, respectively) belonged to chromogenic agar. Conclusion: Chromogenic agar provides more reliable data for S. aureus isolates, and the combined use of all three methods does not significantly increase reliability. doi: https://doi.org/10.12669/pjms.37.5.4274 How to cite this:Gorgun S, Isler H, Turgut MC. Comparison of rapid and conventional methods for investigating of mecA presence in Staphylococcus Species. Pak J Med Sci. 2021;37(5):---------. doi: https://doi.org/10.12669/pjms.37.5.4274 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

scholarly journals The frequency of resistance to antibiotics of most frequently isolated bacteria from blood cultures during the period 1997-2002

2004 ◽  
Vol 61 (4) ◽  
pp. 391-397
Author(s):  
Veljko Mirovic ◽  
Branka Tomanovic ◽  
Sonja Konstantinovic
Keyword(s):  
Clinical Laboratory ◽  
Blood Cultures ◽  
Diffusion Method ◽  
National Committee ◽  
Disk Diffusion Method ◽  
Beta Lactamases ◽  
Resistance To Antibiotics ◽  
Extended Spectrum Beta Lactamases ◽  
High Level

The aim of this study was to determine the frequency of resistance to antibiotics of the most frequently isolated bacteria from blood cultures of hospitalized patients during the period 1997-2002. The resistance to antibiotics was determined by disk diffusion method according to National Committee for Clinical Laboratory Standards procedures. The majority of staphylococci isolates were resistant to methicillin, and the proportion of methicillin-resistant Staphylococcus aureus was stable (76.8-81.6%), during the follow-up period. None of the staphylococci isolates were resistant to vancomycin, but there was a very high incidence of high-level resistance of enterococci to aminoglycosides (47.2-72.2%). In 1998, only one strain among enterococci was resistant to vancomycin (Enterococcus faecium, VanA fenotype). Enterococcus spp isolates expressed variable frequency of resistance to ampicillin (15-40.1%) during the follow-up period. Among Enterobacteriaceae there were no isolates resistant to imipenem, but dramatic increase of the resistance to ceftriaxone was found from 35.9% in 1997 to 95.9% in 2002 (p<0.001). Extended spectrum beta-lactamases production was found in all the species of enterobacteria isolates. Resistance to imipenem was observed in Acinetobacter spp isolates in 2002 for the first time. Pseudomonas spp isolates expressed high and very variable resistance to all antibiotics tested during the follow-up period.

scholarly journals Comparison of Three Methods for Testing Azole Susceptibilities of Candida albicans Strains Isolated Sequentially from Oral Cavities of AIDS Patients

1998 ◽  
Vol 36 (6) ◽  
pp. 1578-1583 ◽  
Author(s):  
Anna Maria Tortorano ◽  
Maria Anna Viviani ◽  
Francesco Barchiesi ◽  
Daniela Arzeni ◽  
Anna Lisa Rigoni ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Clinical Laboratory ◽  
Reference Method ◽  
National Committee ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Aids Patients ◽  
Testing Procedures ◽  
Broth Microdilution Method ◽  
Mean Differences

Three susceptibility testing procedures were compared to determine fluconazole, itraconazole, and ketoconazole MICs against 47Candida albicans strains isolated sequentially from the oral cavities of five AIDS patients undergoing azole therapy. They included the broth microdilution method (BM), performed according to the National Committee for Clinical Laboratory Standards’ tentative standard, the agar dilution method (AD), and the Etest; the latter two tests were performed both in Casitone agar (AD-Cas and Etest-Cas) and in RPMI (AD-RPMI and Etest-RPMI). Twenty-four- and 48-h MICs obtained by AD and Etest were compared with 48-h MICs obtained by BM. The MICs of all the azoles determined by BM were usually lower than those obtained by the other methods, mainly due to different reading criteria. In order to assess the most appropriate way of evaluating the agreement of MICs obtained by different methods with those produced by the proposed reference method (BM), we used the mean differences calculated according to Bland and Altman’s method. Comparison of fluconazole MICs obtained by BM and AD-Cas yielded a mean difference of 3, and the percentages of agreement within ±2 dilutions were 98 and 100% at 24 and 48 h, respectively. For ketoconazole and itraconazole MICs, lower mean differences were noted, and agreement ranged from 96 to 100%. Agreement between the AD-RPMI and BM results was poor for all azoles, and an increase in MICs was always observed between the 1st- and 2nd-day readings. Similarly, Etest-Cas gave better agreement with BM than did Etest-RPMI for all the azoles. BM, AD-Cas, and Etest-Cas each demonstrated a progressive increase in fluconazole MICs against strains isolated sequentially from a given patient, in accordance with the decreased clinical response to fluconazole.

scholarly journals Comparison of methods for the determination of antimicrobial resistance in Campylobacter spp. human and the food chain isolates

2008 ◽  
Vol 52 (No. 4) ◽  
pp. 169-174
Author(s):  
M. Holasova ◽  
R. Karpiskova ◽  
S. Karpiskova ◽  
V. Babak ◽  
J. Schlegelova
Keyword(s):  
Nalidixic Acid ◽  
Antimicrobial Agents ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method ◽  
Microdilution Method ◽  
Study Results ◽  
Agar Dilution

With a microdilution method, using the commercial diagnostic test Sensititre Susceptibility Plates for Campylobacter MIC (Trek Diagnostic Systems, Cleveland, OH, USA), disk diffusion and agar dilution method, resistance to six antimicrobial agents were examined in a reference strain <i>Campylobacter jejuni</i> ATCC 33560 and 73 thermo-tolerant isolates of <i>Campylobacter</i> spp. For the microdilution method and all tested antimicrobial agents, our determined values of microbiological breakpoints of resistant strains were suggested as the minimum inhibitory concentration (MIC<sub>R</sub>) for ciprofloxacin &ge; 0.5, erythromycin &ge; 4, gentamicin &ge; 4, nalidixic acid &ge; 32 and tetracycline &ge; 4 &mu;g/ml. On the basis of our study results, strains resistant to clindamycin were MIC<sub>R</sub> &ge; 2 &mu;g/ml for the dilution methods and a zone diameter R ≤ 16 mm for the disk diffusion method. Comparison of the results of the resistance examination, a microdilution method and disk diffusion method with the reference agar dilution method, showed that all compared methods yielded identical results with the exception of the resistance determination in erythromycin and nalidixic acid. The errors were mostly the result of the interpretation criteria for MIC<sub>R</sub> of agar dilution method and different conditions of cultivation used. However, the compared methods, provide results comparable with the reference method having greater convenience of measurement.

scholarly journals Inaccuracy of the Disk Diffusion Method Compared with the Agar Dilution Method for Susceptibility Testing of Campylobacter spp.

2011 ◽  
Vol 50 (1) ◽  
pp. 52-56 ◽  
Author(s):  
Mirva Lehtopolku ◽  
Pirkko Kotilainen ◽  
Pauli Puukka ◽  
Ulla-Maija Nakari ◽  
Anja Siitonen ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method ◽  
Agar Dilution ◽  
Campylobacter Spp

scholarly journals In vitro efficacy of synergistic antibiotic combinations in imipenem resistant Pseudomonas aeruginosa strains

2017 ◽  
Vol 9 (1) ◽  
pp. 3-8
Author(s):  
Aleya Farzana ◽  
S. M. Shamsuzzaman
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Considerable Proportion ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method ◽  
New Antibiotics

The increase in antibiotic resistance coincided with the decline in production of new antibiotics. Combination antibiotic treatment is preferred in nosocomial infections caused by multidrug resistant Pseudomonas aeruginosa. In vitro synergism test by agar dilution method were used to choose the combinations which might be used in clinic. The aim of this study was to investigate the synergistic efficacy of antibiotic combinations in imipenem resistant P. aeruginosa strains. Carbapenem resistance (imipenem and meropenem) wasdetermined by disk diffusion method. Among isolated P. aeruginosa 44.9% were cabapenem resistant. The MIC of drugs among 25 imipenem resistant isolates ranged from >_ 256 mg/L to <_ 8 mg/L for imipenem, >_ 1024 mg/L to <_ 64 mg/L for ceftriaxone, >_ 256 mg/L to <_ 8 mg/L for amikacin, >_ 16 mg/L to <_ 2 mg/L for colistin, >_ 512 mg/L to <_ 16 mg/L for piperacillin/tazobactam. Among antibiotic combinations, piperacillin /tazobactam- amikacin was most effective with 80% synergism next to which was imipenem-amikacin with 60% synergism, then imipenem-colistin with 50% synergism, imipenem-ceftriaxone with 30% synergism. Only one combination (piperacillin/tazobactum -imipenem showed 20% antagonism. All these combinations had considerable proportion of additive effect which is also desirable for these multi drug resistant isolates.Bangladesh J Med Microbiol 2015; 9 (1): 3-8

scholarly journals Correlation of Oxacillin MIC with mecAGene Carriage in Coagulase-Negative Staphylococci

2000 ◽  
Vol 38 (2) ◽  
pp. 752-754 ◽  
Author(s):  
Zafar Hussain ◽  
Luba Stoakes ◽  
Viki Massey ◽  
Deb Diagre ◽  
Viivi Fitzgerald ◽  
...  
Keyword(s):  
Staphylococcus Epidermidis ◽  
Clinical Laboratory ◽  
National Committee ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Coagulase Negative Staphylococci ◽  
Methicillin Resistant ◽  
Mueller Hinton ◽  
Mueller Hinton Agar ◽  
Agar Dilution

The National Committee for Clinical Laboratory Standards has recently changed the oxacillin breakpoint from ≥4 mg/liter to ≥0.5 mg/liter to detect methicillin-resistant coagulase-negative staphylococci (CoNS) because the previous breakpoint lacked sensitivity. To determine the correlation between the new oxacillin breakpoint and the presence of themecA gene, 493 CoNS of 11 species were tested. The presence of the mecA gene was determined by PCR, and oxacillin susceptibility was determined by the agar dilution method with Mueller-Hinton agar containing 2% NaCl and oxacillin (0.125 to 4.0 mg/liter). The new breakpoint correctly classified all CoNS strains with mecA as methicillin resistant and strains ofStaphylococcus epidermidis, S. haemolyticus, and S. hominiswithout mecA as methicillin susceptible. The breakpoint of ≥0.5 mg/liter was not specific for S. cohnii, S. lugdunensis, S. saprophyticus, S. warneri, and S. xylosus, in that it categorized 70 of 74 strains of these species withoutmecA (94.6%) as methicillin resistant. The results of this study indicate that the new oxacillin breakpoint accurately identifies strains of CoNS with mecAbut is not specific for strains of certain species of CoNS withoutmecA.

scholarly journals Fluconazole Susceptibilities of Bloodstream Candidasp. Isolates as Determined by National Committee for Clinical Laboratory Standards Method M27-A and Two Other Methods

1999 ◽  
Vol 37 (7) ◽  
pp. 2197-2200 ◽  
Author(s):  
Emilia Cantón ◽  
Javier Pemán ◽  
Alfonso Carrillo-Muñoz ◽  
Ana Orero ◽  
Pedro Ubeda ◽  
...  
Keyword(s):  
Clinical Laboratory ◽  
Geometric Mean ◽  
Inhibition Zone ◽  
Diffusion Method ◽  
National Committee ◽  
Dilution Method ◽  
Inhibition Zone Diameter ◽  
Zone Diameter ◽  
The Mean

The in vitro activity of fluconazole against 143Candida spp. obtained from the bloodstreams of 143 hospitalized patients from 1995 to 1997 was studied. Susceptibility tests were carried out by two macrodilution methods, the M27-A and a modified M27-A method (0.165 M, pH 7/morpholinepropanesulfonic acid-buffered RPMI 1640 medium supplemented with 20 g ofd-dextrose per liter), and by the agar diffusion method (with 15-μg fluconazole [Neo-Sensitab] tablets). With 2 μg of fluconazole per ml, 96.92% of 65 C. albicans isolates, 86.2% of 58 C. parapsilosis isolates 7 of 8 C. tropicalis isolates, and 1 of 6 C. glabrata isolates were inhibited. Only one strain of C. albicans and one strain of C. tropicalis were resistant. The agreement between the two macrodilution methods was greater than 90% within ±2 log2 dilutions for all strains except C. glabrata (83.3%) and C. tropicalis(87.5%). Generally, MICs were 1 log2 dilution lower in glucose-supplemented RPMI 1640 medium. No correlation between zone sizes and MICs was found. All strains susceptible by the diffusion test were susceptible by the dilution method, but the converse was not necessarily true. Interestingly, inhibition zones were smaller forC. albicans, for which the geometric mean MIC was 0.29 μg/ml and the mean inhibition zone diameter was 25.7 mm, while for C. parapsilosis the geometric mean MIC was 0.96 μg/ml and the mean inhibition zone diameter was 31.52 mm. In conclusion, the two macrodilution methods give similar results. The modified M27-A method with 2% dextrose has the advantage of shortening the incubation time and simplifying the endpoint determination.

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