Interlaboratory variation of antibiograms of methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains with conventional and commercial testing systems.

Laboratory-prepared (conventional) and commercial susceptibility testing systems were compared by using a group of methicillin-resistant (MR) and methicillin-susceptible (MS) strains of Staphylococcus aureus. A group of 25 MR and 15 MS S. aureus strains were coded and tested blindly by disk diffusion, agar dilution, broth microdilution, Sensititre, Micro-Media, Sceptor, API 3600S, MicroScan, Autobac I, and MS-2 systems. All systems were incubated at 35 degrees C and read with either a manual or automated reader at the recommended times. Where applicable, systems were also read at 48 h. Among the conventional assays, the broth and agar dilution methods were comparable, both detecting 88% of the MR strains at 24 h and detecting 92 and 96%, respectively, at 48 h. The disk diffusion method was less efficient, detecting only 36 and 72% at 24 and 48 h, respectively. Detection of cephalothin resistance was low for all systems at both time periods, with agar dilution and disk diffusion being the most and least efficient, respectively. Some variability was also seen with detection of resistance to clindamycin and gentamicin. Among the MS strains, variability among the conventional systems occurred with methicillin, gentamicin, ampicillin, and penicillin. Comparison of the commercial systems with manual readers with the broth microdilution method (reference method) showed that for MR strains, the Sceptor system gave identical results at 24 and 48 h. Sensititre detected 68 and 88% of the MR strains, whereas Micro-Media was least effective detecting 12 and 80% at 24 and 48 h, respectively. None of the commercial systems detected cephalothin resistance well, with only one strain being indicated by the Sceptor and Sensititre systems at 48 h. Slight differences were also seen among the systems with clindamycin and gentamicin. With regard to the MS strains, variability among the systems was seen with methicillin, penicillin, ampicillin, clindamycin, and gentamicin. Among commercial systems with automated readers, the API system detected a greater number of MR strains than did the reference method at 24 and 48 h, 96 and 100%, respectively. The MicroScan method was comparable to the reference method detecting 80 and 88% of the MR strains at both time periods, respectively. Both Autobac I and MS-2 were much less effective in detecting MR strains, noting only 32 and 16%, respectively, at the 3- to 6-h readings. Poor detection of cephalothin resistance among MR strains was evident in all systems. Variability also occurred among the systems with clindamycin, gentamicin, and ampicillin. A single strain of the MR group was reported to be vancomycin resistant by the API system. Among the MS group, the greatest variability was seen with methicillin. Less variability occurred with penicillin, ampicillin, gentamicin, and vancomycin.     

Serum bactericidal testing with the Autobac system.

Current methodology for the serum bactericidal test requires a minimum of 48 h. A procedure was devised for performing this test with the Autobac system (General Diagnostics, Div. Organon Inc., Raleigh, N.C.) in a shortened time span. All titers obtained with the Autobac were compared against results obtained with a standardized tube dilution procedure. The Autobac low-thymidine eugonic broth performed comparably to the tube dilution diluent, a 1:1 ratio of pooled human serum and cation-supplemented Mueller-Hinton broth (99.2% correlation between bactericidal endpoints). Over 300 tests were conducted by using stock reference bacterial strains, clinical isolates, pooled human serum seeded with antimicrobial agents, and serum from patients on antimicrobial therapy. With the Autobac procedure, serum inhibitory titers can be reported in 3 to 4 h (93.4% correlation with the tube dilution procedure). Serum bactericidal titers can be obtained in 24 h without the necessity of subculturing (95.6% correlation). With the exception of staphylococci tested against penicillin, serum bactericidal titers can be obtained in 3 to 4 h (88.4% correlation). The Autobac procedure can provide the clinical laboratory with a rapid, reliable method for performing the serum bactericidal test.     

Detection of group A streptococcal antigen directly from throat swabs with a ten-minute latex agglutination test.

Using 490 strains of Staphylococcus aureus divided into methicillin-susceptible, -resistant, and -heteroresistant varieties, we compared the results obtained by the agar disk method with those obtained with the automated Autobac system. Susceptible strains exhibited a perfect correlation, whereas there were numerous discrepancies with resistant and still more with heteroresistant varieties. When incubation was increased to 18 h at 37 degrees C (Autobac incubation temperature), 35 degrees C, or 30 degrees C, these differences disappeared, but other problems may arise when incubation is prolonged, especially with erythromycin. We thus recommend carrying out two readings, a normal one after 3 h of incubation and a special reading after 18 h, solely for the detection of heteroresistance to methicillin.     

Evaluation and optimization of urine screening by Autobac.

The purpose of this investigation was to evaluate the effectiveness of the Autobac (Pfizer Inc., New York, N.Y.) urine screen for detection of bacteriuria in 3,026 urine specimens and to establish the optimum procedure for the Autobac system. Overall, 97% of urine specimens having greater than 10(5) colony-forming units (CFU) per ml were detected within 5 h by the Autobac system. The system detected 66, 90, and 94% of such specimens after 2, 3, and 4 h of incubation, respectively. Of specimens having 10(4) to 10(5) CFU/ml, the Autobac system detected 10, 45, 53, and 95% after 2, 3, 4, and 5 h of incubation, respectively. The rate of false-positive results increased from 0% after 2 h to 2% after 3 h to 6% after 4 h and 25% after 5 h of incubation. The specificity of the urine screening results also varied with the incubation time. Percentages of specimens having greater than 10(5) CFU/ml that gave positive urine screening results at various times were as follows: 96% at 2 h, 74% at 3 h, 29% at 4 h, and 9% at 5 h. These findings suggest that a 3- or 4-h urine screening procedure will effectively detect bacteriuria of greater than 10(5) CFU/ml, with few false-positive results. However, a 5-h procedure, which gives more false-positive results, may be needed for detection of lower levels of bacteriuria.     

Variation in the abilities of automated, commercial, and reference methods to detect methicillin-resistant (heteroresistant) Staphylococcus aureus.

The abilities of commercial MIC, automated, and reference methods for in vitro detection of methicillin-resistant Staphylococcus aureus were determined on 49 strains from eight hospitals. Micro-Media, MicroScan, Sensititre, Sceptor, API Uniscept KB, Abbott MS-2, Vitek AMS, Autobac MTS, NCCLS disk diffusion, and broth microdilution antimicrobial susceptibility testing procedures were evaluated. All testing was performed by using manufacturers' or reference procedures, and results were determined at no later than 24 h of incubation at 35 degrees C. With NCCLS disk diffusion, all strains were resistant to oxacillin (1 microgram), 47 (96%) were resistant to methicillin (4 micrograms), and 48 (98%) were resistant to nafcillin (1 microgram). The percentages of strains resistant to methicillin (greater than 8 micrograms/ml) were 98% with API Uniscept KB, 86% with Sceptor, MicroScan, and Autobac MTS, 84% with Sensititre, 71% with Micro-Media, and 70% with NCCLS MIC. Abbott MS-2 detected 86% of strains resistant to methicillin (greater than 5 micrograms/ml). With oxacillin (greater than 2 micrograms/ml), 90% were detected with Vitek AMS and 70% were detected with NCCLS MIC. With nafcillin (greater than 2 micrograms), 82% were resistant with Micro-Media, 57% were resistant by NCCLS MIC, and 50% (three of six) were resistant by MicroScan. Two strains from one hospital and one strain from another gave susceptible results with all automated and commercial methods. All strains from three centers were detected by all methods. Variability also occurred among the systems with cephalothin, clindamycin, gentamicin, chloramphenicol, and trimethoprim sulfamethoxazole.     

Rapid MIC testing with the sensititre autoreader.

Automated microdilution MIC results, obtained with the Autoreader (Sensititre, Inc., Salem, N.H.) following 5 h of incubation, were compared with manually read, concurrent control MICs following 18 h of incubation in a three-laboratory comparative study. A total of 704 members of the family Enterobacteriaceae or similar gram-negative organisms were tested against 17 antimicrobial agents. Autoreader MICs were within 1 doubling dilution of control values in 92.9% of instances. Discrepancies of +/- 2 doubling dilutions and +/- 3 or greater doubling dilutions were noted in 4.5 and 2.6%, respectively, of the 7,687 drug-organism combinations analyzed. The majority of errors occurred when beta-lactam antimicrobial agents were tested with a variety of different species. MICs at 5 h, when Pseudomonas aeruginosa was used, were possible in only half the isolates tested and yielded data on only a limited number of drugs in the remaining instances. Excluding results obtained with penicillin and ampicillin, which were uniformly poor, Staphylococcus aureus Autoreader values were within +/- 1 doubling dilution of control values in 93.6% of instances, 5.4% varied by +/- 2 dilutions, and only 1% of test values by +/- 3 or more dilutions from control values for 82 isolates tested against nine antimicrobial agents. Of eight additional S. aureus isolates tested that were resistant to methicillin, only one was read correctly by the Autoreader, with results on the remaining seven appearing as either insufficient growth or as total resistance to all drugs tested. Interlaboratory reproducibility was excellent for selected isolates of S. aureus and gram-negative bacilli. The accuracy of the Sensititre Autoreader MIC results was comparable to that of other same-day quantitative systems for members of the family Enterobacteriaceae and S. aureus, while the economic and procedural advantages of the broth microdilution method was maintained.     

Detection of methicillin-resistant Staphylococcus aureus by microdilution and disk elution susceptibility systems

To determine whether methicillin-resistant (MR) Staphylococcus aureus from different geographic areas are detected reliably by various commercially available microdilution broth and disk elution systems, 73 such isolates obtained from hospitals in 13 cities were tested by a reference method (agar dilution) and by the Microscan, API 3600S, Autobac I, and MS-2 systems. Both Eugonic broth and Low Thymidine Eugonic broth were used in the evaluation of the Autobac I, and two versions of the MS-2 were used. The proportions of isolates categorized as MR by the various methods were: agar dilution method, 99%; Microscan, 100% (if the suggested cut-off of the manufacturer was used); API 3600S, 96%; Autobac I, 84 to 93%; and MS-2, 54 to 68%. With the MS-2 system, isolates from Jackson, Miss., were classified as susceptible to methicillin more often than were strains from other cities. With the Autobac I (Eugonic broth), only 55% of isolates from Houston, Tex., were classified as MR, whereas 89% of isolates from all other cities were correctly classified as MR. With the API 3600S, strains from some cities were categorized as nafcillin susceptible, whereas strains from other cities were classified as resistant to nafcillin. The results of this study suggest that future evaluations of antimicrobial susceptibility testing systems should include MR strains of S. aureus from several geographic areas.     

Detection of methicillin-resistant Staphylococcus epidermidis.

To determine whether methods suggested for detecting methicillin-resistant Staphylococcus aureus apply equally to methicillin-resistant Staphylococcus epidermidis, 135 S. epidermidis isolates were tested by the Vitek AMS gram-positive susceptibility card (Vitek Systems, Inc., Hazelwood, Mo.) and by modifications of agar screen, disk diffusion, and microdilution methods. Modifications included 24- versus 48-h incubation, unsupplemented versus 2% NaCl-supplemented broth, and standard versus direct inoculum. At 24 h, the highest number of resistant strains, 59, was detected by oxacillin (1 microgram) disk diffusion. At 48 h, three additional strains were judged resistant. With one exception, results for oxacillin disk diffusion and agar screen were equivalent at 24 and 48 h. Vitek detected 50 resistant strains. Significantly fewer resistant strains were detected at 24 h by methicillin disk diffusion (5 micrograms) and methicillin microdilution with 2% NaCl. For oxacillin microdilution, neither 2% NaCl supplementation nor the method of inoculum preparation significantly affected the results. Oxacillin microdilution with cation- rather than non-cation-supplemented broth detected significantly fewer (n = 33) resistant strains at 24 h; 51 were resistant at 48 h. To detect methicillin-resistant S. epidermidis, a direct inoculum with either 24-h oxacillin disk diffusion and reincubation of intermediate strains for an additional 24 h or 24-h oxacillin agar screen and reincubation of strains with no growth for a total of 48 h is recommended.     

Specific identification of Staphylococcus aureus by Staphychrom II, a rapid chromogenic staphylocoagulase test

We compared the performance of Staphychrom II (International Microbio, Signes, France), a rapid (2-h) chromogenic staphylocoagulase test that uses human prothrombin and protease inhibitors, with those of the reference tube coagulase test (TCT) and the latex agglutination test (LAT) Slidex Staph Plus for the rapid identification of S. aureus. Prospective evaluation with 293 fresh clinical isolates yielded sensitivities, specificities, and predictive and negative predictive values of 98.1, 100, 100, and 95.1%, respectively, for the Staphychrom II test; 98.6, 98.7, 99.6, and 96.3%, respectively, for LAT; and 97.6, 98.7, 99.5, and 93.9%, respectively, for TCT. The perfect specificity of the Staphychrom II test was confirmed by testing 193 collection strains selected because of their potential testing pitfalls. The Staphychrom II test was positive for 90% of the 215 S. aureus strains tested after only 1 h of incubation. The Staphychrom II test was as sensitive as the reference TCT and was 100% specific.     

Determination of the range of antibacterial activity by use of viable counts

The activity of three aminoglycosides and six beta-lactam antibiotics on strains of Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, and enterococci was studied. The minimal inhibitory concentrations (MICs), the minimal bactericidal concentrations (MBCs), and the minimal antibiotic concentrations (MACs) were determined after 5 h of incubation in broth cultures by colony-forming-unit counts. The MICs were also determined by agar dilution after 24 h of incubation. The MICs on agar after 24 h of incubation were higher than those in broth after 5 h of incubation. The differences ranged from 1.1- to 14.2-fold, but in most cases were only three- to fivefold (P less than 0.05 to less than 0.001). The MBCs at 5 and 24 h were comparable in 71% of tests. For current practice, the MBC of enterococci can be determined after 5 h of incubation with antibiotics. The aminoglycosides showed MBCs which were closer to the MICs than were those of the beta-lactam antibiotics, which required a higher multiple of the MIC to show a bactericidal effect. The MBCs of oxacillin and cefamandole for S. aureus after 5 h of incubation were greater than 128 times the respective MICs. The MACs ranged from 1/1.5 to 1/7 of the 5-h MICs. The three endpoints, MIC, MBC, and MAC, indicate the antibacterial range of an antibiotic in terms of inhibition of growth and bacterial survival. The data suggest that the antibacterial range of an antibiotic is similar for most strains of a given species and is, to some extent, a characteristic of similar antibiotics.     

Collaborative clinical evaluation of the Autobac IDX system for identification of gram-negative bacilli

The Autobac IDX system (General Diagnostics, Warner-Lambert Co., Morris Plains, N.J.) for rapid, semiautomated identification of gram-negative bacilli was compared with the identification methods in routine use in four laboratories. The study included 1,515 organisms representing 30 species of enteric and nonenteric bacteria. Discrepancies between the results of the IDX system and routine methods were resolved by classical biochemical testing at a reference center. Overall, 98% of the organisms were correctly identified by the routine methods, and 93% were correctly identified by the IDX systems. After adjustment for frequency of clinical occurrence of the organisms tested, the IDX system performed with 95% accuracy. Results with the IDX system were available in 3 to 6 h. Results with the comparative methods were available in 4 to 48 h. A wide variety of organisms, including oxidase positive, oxidase negative, fermentative, and nonfermentative, were identified by a single system by using Autobac. Three or more systems were required to identify the 30 species by the comparative methods. Overall, the results indicate the Autobac IDX system is useful for the rapid identification of enteric and nonfermentative gram-negative bacilli.     

In vitro antimicrobial susceptibility testing of Borrelia burgdorferi: a microdilution MIC method and time-kill studies.

The susceptibility of Borrelia burgdorferi, the causative agent of Lyme borreliosis, to various antimicrobial agents varies widely among published studies. These differences are probably due in part to variations in susceptibility testing techniques and growth endpoint determinations. We developed a microdilution method for determining the MICs of antibiotics against B. burgdorferi. The method incorporated BSK II medium, a final inoculum of 10(6) cells per ml, and a 72-h incubation period and was found to be simple and highly reproducible. A variety of antibiotics and strains of B. burgdorferi and one strain of Borrelia hermsii were examined by this method. MICs of penicillin, ceftriaxone, and erythromycin for the B31 strain of B. burgdorferi were 0.06, 0.03, and 0.03 microgram/ml, respectively. We compared the MICs obtained by the microdilution method with those obtained by a macrodilution method using similar criteria for endpoint determinations and found the values obtained by both methods to be in close agreement. To further investigate the bactericidal activities of penicillin, ceftriaxone, and erythromycin against strain B31, we used subsurface plating to determine MBCs and we also performed time-kill studies. The MBCs of penicillin, ceftriaxone, and erythromycin were 0.125, 0.03, and 0.06 micrograms/ml, respectively. Time-kill curves demonstrated a greater than or equal to 3-log10-unit killing after 72 h with penicillin, ceftriaxone, and erythromycin; ceftriaxone provided the greatest reduction in CFU. The described methods offer a more standardized and objective approach to susceptibility testing of B. burgdorferi.     

Rapidly increasing prevalence of penicillin-resistant Streptococcus pneumoniae in middle Tennessee: a 10-year clinical and molecular analysis

The clinical and molecular epidemiology of penicillin-resistant Streptococcus pneumoniae and the diagnostic accuracy of a six-primer PCR assay in identifying penicillin resistance were analyzed by using clinical isolates recovered over a 10-year period in middle Tennessee. The prevalence of non-penicillin-susceptible S. pneumoniae isolates (MIC, > or =0.1 microg/ml) increased from 10% in 1990 to 70% in 1999 (P < 0.001). Among S. pneumoniae isolates for which the penicillin MIC was > or =2 microg/ml (highly penicillin-resistant S. pneumoniae [PRSP]), 23 and 5% were resistant to at least three and at least five other antimicrobial classes, respectively. Pulsed-field gel electrophoresis identified 13 unique strain types, with type B accounting for 33% of PRSP isolates. The sensitivity, specificity, and negative and positive predictive values of the PCR assay in detecting PRSP were 99, 100, 99, and 100%, respectively. Penicillin resistance is rapidly increasing among S. pneumoniae isolates in Tennessee. The simultaneous detection of S. pneumoniae and high-level penicillin resistance can be accurately performed with the six-primer PCR assay.     

广州地区老年人肺炎链球菌临床分离株的青霉素耐药研究与分子分型

目的调查广州地区老年患者肺炎链球菌分离株对青霉素的敏感性，并分析其亲缘关系。方法K—B纸片法对33株分离自老年住院病人的肺炎链球菌进行青霉素药敏试验；应用PCR技术检测青霉素结合蛋白基因pbp1a，pbp2x，pbp2b；用盒式PCR（BOX—PCR）分析菌株间亲缘关系。用多位点测序分型技术（multilocus sequence typing，MLST）检测青霉素耐药菌株的分子分型。结果青霉素的耐药率为3．03％（1／33）：用PCR方法鉴定PSSP的准确率为68．75％；BOX-PCR可将这33株肺炎链球菌分为21型。MIST分型显示，青霉素耐药菌株属ST271型。结论广州地区老年患者肺炎链球菌对青霉素耐药率较低．用PCR方法检测PSSP有一定的可行性。BOX—PCR显示了较高的分辨率，能快速可靠地检测菌株间的亲缘关系。广州地区流行的耐药克隆与Taiwan^19F-14株同源。     

A novel luminometer for rapid antimicrobial susceptibility tests on gram-positive cocci by ATP bioluminescence

The susceptibility of 130 clinical isolates of gram-positive cocci to a wide range of antimicrobial agents was assessed by ATP bioluminescence in a 4-h test. ATP assays were performed on a novel luminometer, the Amerlite Analyser, which measures luminescence from microtitration trays. For most organisms tested, there was good correlation (greater than 90%) with conventional MIC values estimated on 18-h cultures. However, a problem was found with detection of penicillin resistance in Staphylococcus aureus by the ATP method, 13% of strains showing major disagreement. Methicillin resistance of S. aureus was shown reliably for most strains (94%) by ATP assay, provided they were incubated at 30 degrees C. The Amerlite Analyser offers the potential for the development of a semi-automated antimicrobial susceptibility test, with a significant reduction in reagent costs when compared with previously described bioluminescence protocols.     

In vivo bactericidal activity of an oxacillin-netilmicin combination against Staphylococcus aureus. Influence of the rhythm of netilmicin administration

The bactericidal activity of two regimens of netilmicin (8 mg/kg/day) given intravenously once a day (od) or thrice daily (tid) both alone and in combination with oxacillin (200 mg/kg/day) was compared using a model of fibrin clots infected with a strain of Staphylococcus aureus (10(7) CFU/g) sensitive to methicillin and netilmicin (clinical isolate) and implanted subcutaneously in rabbit. This study shows that: 1) Netilmicin given alone as both single and divided doses results in early bacterial killing but does not exert a bactericidal effect after 24 hours because of a significant late increase of the number of bacterial. 2) The netilmicin-oxacillin combinations are more bactericidal at 1 h, 2 h and 24 h than oxacillin alone (P less than 0.001). 3) The oxacillin-netilmicin combination appears to be better for bacterial killing when netilmicin is given thrice daily (P less than 0.001). It is hard to draw a clinical inference from such an experimental study but it seems that 8-hour divided doses intervals should be better for administration of netilmicin than single daily dose during the acute period of staphylococcal infections.     

Problem of the in vitro sensitivity to cefamandole of methicillin-resistant Staphylococcus aureus

In work involving 127 strains of methicillin resistant Staphylococcus aureus, belonging to two resistance categories, homogeneous or RO and heterogeneous or RH and using five cephalosporins, cefalotin, cefazolin, cefamandole, cefotaxime and latamoxef, the authors compared the results of their in vitro sensitivity measurements with two methods: agar disk diffusion and growth inhibition in liquid medium (Autobac). They also compared the MIC values with a significant fraction (19 strains). In order to detect heterogeneous resistance experiments were carried out in duplicate, with one series grown in hypertonic medium and incubated for 48 heures at 37 degrees C, or grown on normal medium with lower incubation at 30 degrees C. In both defined techniques and in vitro, it was observed that the 127 methicillin resistant Staphylococcus aureus strains except one were sensitive to cefamandole and that 26 were also sensitive to cefalotin. Considerable caution is necessary in the in vivo transposition of these findings. May be present media are not able to allow growth of resistant strains to cefamandole.     

Comparison of genes involved in penicillin resistance in staphylococci of bovine origin

Ten penicillin-resistant and -susceptible staphylococci, isolated from bovine mastitis milk, were studied for the presence of genes that are, or may be, involved in resistance against penicillin. The repressor (blaI), antirepressor (blaR1), and structural (blaZ) genes of the beta-lactamase-operon were found to be closely linked in all penicillin-resistant strains. The beta-lactamase gene cluster was more commonly located on chromosomal rather than plasmid DNA in the strains studied. The transposase (p480) gene, which has been identified in the Staphylococcus aureus beta-lactamase transposon Tn552, was found in only one single penicillin-resistant S. aureus strain. The other penicillin-resistant S. aureus isolates contained IS1181 in close location with the beta-lactamase gene cluster. In only one S. haemolyticus isolate was the beta-lactamase gene cluster found in close association with IS257. Penicillin-resistant S. aureus strains, which were additionally resistant to tetracycline, contained IS257 in close association with the tetracycline resistance gene (tetK). Sequence analysis of blaI, blaR1, and blaZ in two penicillin-resistant S. aureus strains revealed 94-96% sequence homology with bla in staphylococci of human origin. The results indicate a predominance of class I bla transposons rather than Tn3 family class II transposons in the isolates used in this study.     

Evaluation of the E test for susceptibility testing of anaerobic bacteria.

The susceptibilities of 105 clinical isolates of anaerobic bacteria were determined by a new method, the E test (AB Biodisk, Solna, Sweden), and were compared with the MICs for these organisms obtained by the reference agar dilution method by using supplemented brucella and Wilkins-Chalgren agars. The E test is a plastic strip with a predefined antibiotic gradient immobilized on one side and a MIC interpretive scale printed on the other side. Strips with cefoxitin, cefotaxime, imipenem, penicillin, metronidazole, and clindamycin were used in this study. A suspension of the test strain equal to the visual turbidity of a no. 0.5 McFarland standard was prepared and swabbed onto a 150-mm-diameter plate. The strips were applied in a radial fashion, and the plates were incubated under anaerobic conditions. After growth had occurred, an ellipse of inhibition was seen around each strip. At the point of intersection of the ellipse with the strip, the MIC was read from the interpretive scale. For most antibiotic-organism combinations, the ellipse was clear and the endpoint was sharp. The E-test MICs were interpreted after overnight and 48-h incubation for 58 of the strains. After overnight incubation, 87% of the E-test MICs were within 1 dilution of the agar dilution MICs, and 98% were within 2 dilutions. After 48 h of incubation, agreement was 86 and 97% respectively. E-test MICs obtained for the Bacteroides fragilis group after overnight incubation were more comparable than those obtained after 48 h of incubation to agar dilution MICs determined at 48 h for all drugs except clindamycin. On brucella agar, there was a 2% categorical discrepancy rate between the E-test MICs and agar dilution MICs, which occurred mostly with cefoxitin. The E test is easy to perform and read, is suitable for all anaerobes, can be used to test single patient isolates as needed, and offers the laboratory a reliable method for susceptibility testing of anaerobic bacteria.     

Antibiotic resistance of Staphylococcus aureus at north Lebanon: place of the methicillin resistance and comparison of detection methods

The aim of this study was to evaluate the susceptibility of 100 Staphylococcus aureus strains isolated from the laboratory of Microbiology of the Islami Hospital of Tripoli (Lebanon) to 19 antibiotics, and to determine the prevalence of methicillin resistant strains. 30% of strains studied were methicillin resistant, 96% were resistant to the penicillin G. Clavulanic acid restaurated the amoxicillin activity to 29%. The resistance level was 34% for amikacin, 3% for gentamycin and tobramycin, 10% for chloramphenicol, 44.33% for tetracyclin, 7% for erythromycin, 4.04% for clindamycin, 20% for trimethoprim-sulfametoxasol and 0% for vancomycin and teicoplanin. The methicillin-resistant Staphylococcus aureus possess more important resistant level in comparison with the methicillin sensitive strains. We compared the ability of latex agglutination test (Slidex(R) SARM, bioMérieux, France) to detect the production of penicillin-binding protein 2' (PBP 2') in 100 clinical isolates of S. aureus with two reference methods: the oxacillin disk diffusion test and the MIC determination by the E-test (AB BIODISK, Sweden). The two reference methods give the same results for the detection of methicillin resistant S. aureus. The Slidex test was positive for all 30 isolates determined to be methicillin resistant by the reference methods (sensitivity 100%). The latex test was negative for 42 of 70 isolates determined to be methicillin susceptible by the reference methods, and the latex test was positive for 28 isolates determined to be susceptible (specificity 60%).