Comparative assessment of inoculum effects on the antimicrobial activity of amoxycillin-clavulanate and piperacillin-tazobactam with extended-spectrum β-lactamase-producing and extended-spectrum β-lactamase-non-producing Escherichia coli isolates.

A significant inoculum-size effect has been observed with piperacillin-tazobactam, and has been associated with β-lactamase production in extended-spectrum β-lactamase (ESBL) producers. This association has not been previously studied in the case of amoxycillin-clavulanate. Piperacillin-tazobactam and amoxycillin-clavulanate were compared, using high inocula of susceptible strains either harbouring ESBLs or not. Two non-ESBL-producing and 15 amoxycillin-clavulanate-susceptible and piperacillin-tazobactam-susceptible ESBL-producing Escherichia coli isolates, and their respective transconjugants, were tested in dilution susceptibility tests using standard and 100-fold higher inocula. Three ESBL-producing strains and E. coli ATCC 25922 were selected for time-kill studies using standard and high initial inocula. At high inocula, MICs of piperacillin increased >eight-fold for non-ESBL-producing strains, and MICs of piperacillin-tazobactam (8 : 1 ratio or with tazobactam fixed at 4 mg/L) increased>eight-fold for all ESBL-producing strains. However, amoxycillin MICs were not affected by a high inoculum with non-ESBL-producing strains, whereas the MICs of amoxycillin-clavulanate (2 : 1 and 4 : 1) increased ≤four-fold for ESBL producers, using the broth and agar dilution methods. In kinetic studies at a high inoculum, amoxycillin and amoxycillin-clavulanate were bactericidal against E. coli ATCC 25922, whereas piperacillin and piperacillin-tazobactam yielded decreases of <1 log₁₀ CFU/mL. Similarly, at a high inoculum, only amoxycillin-clavulanate was able to maintain bactericidal rates of killing over 24 h against the ESBL-positive E. coli isolates. The stability of amoxycillin-clavulanate and the contrasting results obtained with piperacillin-tazobactam against high inocula of ESBL-non-producing and ESBL-producing E. coli strains appear to be related to aspects other than the amount of β-lactamase production.     

Comparison of Rosco Neo-Sensitabs with Oxoid paper disks in EUCAST disk diffusion antimicrobial susceptibility testing on Mueller–Hinton agar

This study compared Neo-Sensitabs with Oxoid paper disks using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) disk diffusion antimicrobial susceptibility test on Mueller–Hinton agar. The EUCAST-recommended quality control strains (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212) (Part I) and clinical isolates (Part II) were investigated. In Part I of the study, 27 combinations of antimicrobial agents were tested on four quality control strains repeatedly up to 60 times and zone diameters of tablets and disks were compared. In Part II of the study, 351 clinical isolates were included to cover a broad range of species, as well as resistance mechanisms. In Part I, four major deviations (>1 mm outside quality control ranges) were observed with Neo-Sensitabs. In one case with P. aeruginosa ATCC 27853 (meropenem), there was a corresponding major deviation (2 mm) with the Oxoid disk. The three remaining major deviations with Neo-Sensitabs were observed with meropenem (2 mm) in E. coli ATCC 25922 and with ciprofloxacin (2 mm) and gentamicin (3 mm) in P. aeruginosa ATCC 27853. For Oxoid disks, there were only minor deviations (=1 mm outside quality control ranges) in these three cases. In Part II, there were six discrepancies, susceptible versus resistant, in 3,533 comparisons between the two methods with the clinical isolates. The Rosco Neo-Sensitabs appear to be a possible alternative to Oxoid paper disks for EUCAST disk diffusion antimicrobial susceptibility testing on Mueller–Hinton agar.     

Automated versus Manual Sample Inoculations in Routine Clinical Microbiology: a Performance Evaluation of the Fully Automated InoqulA Instrument

The process of plate streaking has been automated to improve the culture readings, isolation quality, and workflow of microbiology laboratories. However, instruments have not been well evaluated under routine conditions. We aimed to evaluate the performance of the fully automated InoqulA instrument (BD Kiestra B.V., The Netherlands) in the automated seeding of liquid specimens and samples collected using swabs with transport medium. We compared manual and automated methods according to the (i) within-run reproducibility using Escherichia coli-calibrated suspensions, (ii) intersample contamination using a series of alternating sterile broths and broths with >10⁵ CFU/ml of either E. coli or Proteus mirabilis, (iii) isolation quality with standardized mixed bacterial suspensions of diverse complexity and a 4-category standardized scale (very poor, poor, fair to good, or excellent), and (iv) agreement of the results obtained from 244 clinical specimens. By involving 15 technicians in the latter part of the comparative study, we estimated the variability in the culture quality at the level of the laboratory team. The instrument produced satisfactory reproducibility with no sample cross-contamination, and it performed better than the manual method, with more colony types recovered and isolated (up to 11% and 17%, respectively). Finally, we showed that the instrument did not shorten the seeding time over short periods of work compared to that for the manual method. Altogether, the instrument improved the quality and standardization of the isolation, thereby contributing to a better overall workflow, shortened the time to results, and provided more accurate results for polymicrobial specimens.     

Performance of Copan WASP for Routine Urine Microbiology

This study compared a manual workup of urine clinical samples with fully automated WASPLab processing. As a first step, two different inocula (1 and 10 μl) and different streaking patterns were compared using WASP and InoqulA BT instrumentation. Significantly more single colonies were produced with the10-μl inoculum than with the 1-μl inoculum, and automated streaking yielded significantly more single colonies than manual streaking on whole plates (P < 0.001). In a second step, 379 clinical urine samples were evaluated using WASP and the manual workup. Average numbers of detected morphologies, recovered species, and CFUs per milliliter of all 379 urine samples showed excellent agreement between WASPLab and the manual workup. The percentage of urine samples clinically categorized as positive or negative did not differ between the automated and manual workflow, but within the positive samples, automated processing by WASPLab resulted in the detection of more potential pathogens. In summary, the present study demonstrates that (i) the streaking pattern, i.e., primarily the number of zigzags/length of streaking lines, is critical for optimizing the number of single colonies yielded from primary cultures of urine samples; (ii) automated streaking by the WASP instrument is superior to manual streaking regarding the number of single colonies yielded (for 32.2% of the samples); and (iii) automated streaking leads to higher numbers of detected morphologies (for 47.5% of the samples), species (for 17.4% of the samples), and pathogens (for 3.4% of the samples). The results of this study point to an improved quality of microbiological analyses and laboratory reports when using automated sample processing by WASP and WASPLab.     

Ability of National Committee for Clinical Laboratory Standards-recommended quality control strains from the American Type Culture Collection to detect errors in disk diffusion susceptibility tests.

The National Committee for Clinical Laboratory Standards (NCCLS) recommends, as a quality control for the disk diffusion susceptibility test, the use of three strains from the American Type Culture Collection: Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, and Escherichia coli ATCC 25922. This study assesses the capacity of these strains to detect errors in the overall method. ATCC strains were tested by comparing testing by the standard NCCLS-recommended procedure (ST) with testing under the following conditions: incubation at 25 degrees C, Mueller-Hinton agar depths of 2 mm (AD2) and 8 mm (AD8), agar pHs of 6.5 and 8, inocula with McFarland standards of 0.25 (0.25M) and 4.0 McFarland (4.0M), direct inoculation without preincubation of inoculum (DI), and a 2-h delay between inoculation and disk application (2HR). The frequency of zone measurements outside the NCCLS-recommended control zone limits were as follows: ST, 0%; AD2, 18%; AD8, 9.6%; pH 6.5, 7.9%; pH 8, 5.3%; 0.25M, 3.5%; 4.0M, 24%; DI, 3.4%; 2HR, 1.8%; 25 degrees C (only E. coli and P. aeruginosa were evaluable), 28%. These results suggest that the quality control strains are only partially effective in detecting single extreme laboratory errors and that careful laboratory supervision is necessary even in the setting of properly monitored quality control strains.     

Comparison of automated processing of flocked swabs with manual processing of fiber swabs for detection of nasal carriage of Staphylococcus aureus

The sensitivity of automated culture of Staphylococcus aureus from flocked swabs versus that of manual culture of fiber swabs was prospectively compared using nasal swabs from 867 patients. Automated culture from flocked swabs significantly increased the detection rate, by 13.1% for direct culture and 10.2% for enrichment culture.     

Evaluation of Anatomically Designed Flocked Rectal Swabs for Molecular Detection of Enteric Pathogens in Children Admitted to Hospital with Severe Gastroenteritis in Botswana

Two-hundred eighty matched bulk stool and anatomically designed flocked rectal swab samples were collected from children admitted to the hospital with acute diarrhea in Botswana. Their parents were asked about the acceptability of the swab collection method compared with bulk stool sampling. All samples underwent identical testing with a validated 15-target (9 bacterial, 3 viral, and 3 parasite) commercial multiplex PCR assay. The flocked swabs had a 12% higher yield for bacterial pathogen targets (241 versus 212; P = 0.003) compared with that of stool samples, as well as similar yields for viral targets (110 versus 113; P = 0.701) and parasite targets (59 versus 65; P = 0.345). One hundred sixty-four of the flocked swab-stool pairs were also tested with separate laboratory-developed bacterial and viral multiplex assays, and the flocked rectal swabs had a performance that was similar to that seen with commercial assay testing. Almost all parents/guardians found the swabs acceptable. Flocked rectal swabs significantly facilitate the molecular diagnosis of diarrheal disease in children.     

Determination of Disk Diffusion and MIC Quality Control Guidelines for GSK2140944, a Novel Bacterial Type II Topoisomerase Inhibitor Antimicrobial Agent

GSK2140944 is a novel bacterial type II topoisomerase inhibitor in development for the treatment of conventional and biothreat pathogens, including Gram-positive pathogens and methicillin-resistant Staphylococcus aureus. This quality control study was performed to establish ranges for selected control strains: S. aureus ATCC 29213 and ATCC 25923, Escherichia coli ATCC 25922, Haemophilus influenzae ATCC 49247, and Streptococcus pneumoniae ATCC 49619. The control ranges will be crucial for the accurate evaluation of GSK2140944 potency as it progresses through clinical trial development.     

Antimicrobial Activity of Carpolobia Lutea Extracts and Fractions

Carpolobia lutea (G. Don) (Polygalaceae) is a tropical medicinal plant putative in traditional medicines against gonorrhea, gingivitis, infertility, antiulcer and malaria. The present study evaluated the antimicrobial, antifungal and antihelicobacter effects of extracts C. lutea leaf, stem and root. The extracts were examined using the disc-diffusion and Microplates of 96 wells containing Muller-Hinton methods against some bacterial strains: Eschericia coli (ATCC 25922), E. coli (ATCC10418), Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Staphyllococus aureus (ATCC 6571), Enterococcus faecalis (ATCC 29212) and Bacillus subtilis (NCTC 8853) and four clinical isolates: one fungi (Candida albican) and three bacteria (Salmonella, Sheigella and staphylococcus aureus). The Gram-positive bacteria: Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 29212), Bacillus subtilis (ATCC 19659) and the Gram-negative bacteria: Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Cândida albicans (ATCC 18804) and Helicobacter pylori (ATCC 43504). Some of these extracts were found to be active against some tested strains but activity against H. pylori was >1000mg/ml and good fungistatic activity against C. albican. The MIC against C. albican is in the order n-HF > CHF > ETF= EAF.The order of potency of fraction was the ethanol root > n-HF leaf > ethanol fraction stem > chloroform fraction leaf = ethyl acetate fraction leaf. Polyphenols were demonstrated in ethanol fraction, ethyl acetate fraction, crude ethyl acetate extract and ethanol extract, respectively. These polyphenols isolated may partly explain and support the use of C. lutea for the treatment of infectious diseases in traditional Ibibio medicine of Nigeria.     

Postantibiotic and bactericidal effect of imipenem againstPseudomonas aeruginosa

The postantibiotic effect of imipenem onPseudomonas aeruginosa was studied at different inocula using one ATCC strain and four clinical isolates. The postantibiotic effect was measured using two different methods: viable counts and bioluminescence assay of intracellular bacterial ATP. The postantibiotic effect could be demonstrated with both methods (viable counts 1–2 h, ATP assay 3–5 h) for all strains at an inoculum of 10⁶ CFU/ml. When the inoculum was raised to 10⁸ CFU/ml, no postantibiotic effect could be observed with either method using routine growth conditions. This disappearance of the postantibiotic effect coincided with a loss of bactericidal effect of imipenem when high inocula were used. Improved oxygenation of the cultures restored the bactericidal and postantibiotic effects of imipenem at high inocula.     

Results of Use of WHO Global Salm-Surv External Quality Assurance System for Antimicrobial Susceptibility Testing of Salmonella Isolates from 2000 to 2007

An international External Quality Assurance System (EQAS) for antimicrobial susceptibility testing of Salmonella was initiated in 2000 by the World Health Organization (WHO) Global Salm-Surv in order to enhance the capacities of national reference laboratories to obtain reliable data for surveillance purposes worldwide. Seven EQAS iterations have been conducted from 2000 to 2007. In each iteration, participating laboratories submitted susceptibility results from 10 to 15 antimicrobial agents for eight Salmonella isolates and an Escherichia coli reference strain (ATCC 25922). A total of 287 laboratories in 102 countries participated in at least one EQAS iteration. A large number of laboratories reported results for the E. coli ATCC 25922 reference strain which were outside the quality control ranges. Critical deviations for susceptibility testing of the Salmonella isolates varied from 4% in 2000 to 3% in 2007. Consistent difficulties were observed in susceptibility testing of amoxicillin-clavulanic acid, cefotaxime, ceftazidime, streptomycin, sulfonamides, and tetracycline. Regional variations in performance were observed, with laboratories in central Asia, Africa, and the Middle East not performing as well as those in other regions. Results from the WHO Global Salm-Surv EQAS show that most laboratories worldwide are capable of correctly performing antimicrobial susceptibility testing of Salmonella isolates, but they also indicate that further improvement for some laboratories is needed. In particular, further training and dissemination of information on quality control, appropriate interpretive criteria (breakpoints), and harmonization of the methodology worldwide through WHO Global Salm-Surv and other programs will contribute to the generation of comparable and reliable antimicrobial susceptibility data (D. M. A. Lo Fo Wong, R. S. Hendriksen, D. J. Mevius, K. T. Veldman, and F. M. Aarestrup, Vet. Microbiol. 115:128-139, 2006).     

In vitro activity of FCE 22101, imipenem,and ceftazidime against over 6,000 bacterial isolates and MIC quality control limits of FCE 22101

Six geographically separate laboratories within the USA tested 6,198 bacterial isolates against FCE 22101 (a penem), imipenem (a carbapenem) and ceftazidime (a third-generation cephalosporin). Ninety-three percent of 2,749Enterobacteriaceae were inhibited by FCE 22101, while 95 % were susceptible to ceftazidime and 99 % were susceptible to imipenem. FCE 22101 had little activity againstPseudomonas spp. but was active against most gram-positive pathogens, including enterococci. FCE 22101 MICs for standard quality control strains were defined as 0.5–2.0µg/ml forEscherichia coli ATCC 25922, 2–8µg/ml forEnterococcus faecalis ATCC 29212 and 0.06–0.25µg/ml forStaphylococcus aureus ATCC 29213.     

The influence of killed Propionibacterium granulosum on experimental infection with Escherichia coli

Mice infected subcutaneously with 5 × 10³ viable cells of Escherichia coli ATCC 25922 incorporated in liquified agar developed a systemic infection. Increasing bacterial numbers could be recovered from the liver at several days following infection. Ultimately, the animals died 6 days after infection.Treatment of mice with 1 mg killed Propionibacterium granulosum KP-45 lead to an increased spleen weight at day 7 after intraperitoneal injection. Hence, the animals were highly susceptible to the lethal action of endotoxin. They were, however, markedly protected against infection with E. coli, since definitely lower bacterial counts were found in the liver of pretreated mice in comparison to controls.     

Antibacterial and Anti-Biofilm Activity of Flavonoids and Triterpenes Isolated from the Extracts of Ficus Sansibarica Warb. Subsp. Sansibarica (Moraceae) Extracts

Background

Ficus species are used in African traditional medicine in the treatment of a wide variety of ailments and diseases such as convulsive disorder, wound healing, gonorrhea, tuberculosis, diabetes, diarrhoeal infections, dysentery, malaria and HIV. The aim of this study was to isolate the phytochemical constituents in the plant and test them for their antibacterial activity.

Materials and methods

The fruits, leaves and stem bark were extracted with organic solvents and the compounds in the extracts separated and purified by column chromatography before being identified by NMR spectroscopy and by comparison of the NMR data against values reported in the literature. The antibacterial activity of the pure compounds and extracts were tested using the disk diffusion method.

Results

Three triterpenes and three flavonoids: lupeol acetate (1); cycloart-23-ene-3,25-diol (2); β-sitosterol (3); 5,7,4′-trihydroxyflavan-3-ol (4); epicatechin (5); and isovitexin (6) were isolated in this study. Antimicrobial activity was observed at 8 mg mL⁻¹ for Staphylococcus aureus ATCC 29213 with four of the six isolated compounds, with no activity being observed at 1 – 4 mg mL⁻¹ against Escherichia coli ATCC 25922, E. coli ATCC 35218 and S. aureus ATCC 43300. Epicatechin (5) was found to decrease adhesion of E. coli ATCC 25922 and S. aureus ATCC 29213. Decreased adhesion of S. aureus ATCC 29213 was also observed with 5,7,4′-trihydroxyflavan-3-ol (4) and isovitexin (6).

Conclusions

The results of this study provide baseline information on F. sansibarica''s potential validity in the treatment of infections associated with Gram-positive microorganisms.     

Swabbing for respiratory viral infections in older patients: a comparison of rayon and nylon flocked swabs

The purpose of this study was to compare the sampling efficacy of rayon swabs and nylon flocked swabs, and of oropharyngeal and nasopharyngeal specimens for the detection of respiratory viruses in elderly patients. Samples were obtained from patients 60 years of age or above who were newly admitted to Sorlandet Hospital Arendal, Norway. The patients were interviewed for current symptoms of a respiratory tract infection. Using rayon swabs and nylon flocked swabs, comparable sets of mucosal samples were harvested from the nasopharynx and the oropharynx. The samples were analysed using real-time polymerase chain reaction (PCR) methods. A total of 223 patients (mean age 74.9 years, standard deviation [SD] 9.0 years) were swabbed and a virus was recovered from 11% of the symptomatic patients. Regardless of the sampling site, a calculated 4.8 times higher viral load (95% confidence interval [CI] 1.3–17, p = 0.017) was obtained using the nylon flocked swabs as compared to the rayon swabs. Also, regardless of the type of swab, a calculated 19 times higher viral load was found in the samples from the nasopharynx as compared to the oropharynx (95% CI 5.4–67.4, p < 0.001). When swabbing for respiratory viruses in elderly patients, nasopharyngeal rather than oropharyngeal samples should be obtained. Nylon flocked swabs appear to be more efficient than rayon swabs.     

Detecting Clostridium difficile Spores from Inanimate Surfaces of the Hospital Environment: Which Method Is Best?

The recovery of Clostridium difficile spores from hospital surfaces was assessed using rayon swabs, flocked swabs, and contact plates. The contact plate method was less laborious, achieved higher recovery percentages, and detected spores at lower inocula than swabs. Rayon swabs were the least efficient method. However, further studies are required in health care settings.     

Better Detection of Staphylococcus aureus Nasal Carriage by Use of Nylon Flocked Swabs

Flocked swabs (Copan) were compared to rayon swabs (Copan) for the nasal detection of Staphylococcus aureus in 90 healthy volunteers sampled sequentially during a 5-week period. The use of flocked swabs improved the number of nasal carriers (P = 0.026), the number of positive specimens (P = 0.01), and the quantity of bacteria in positive samples (P = 0.004).Staphylococcus aureus nasal carriage is a major risk factor for S. aureus infection, and the anterior nares are the primary reservoir of S. aureus in humans (19). Therefore, it is desirable to optimize S. aureus detection and to establish the carriage status, particularly in the aim of prophylactically decolonizing patients at risk of infection with this bacterium (4).The detection of S. aureus carriage is usually carried out by nasal sampling. In comparison to the use of rayon swabs, the use of flocked swabs has been demonstrated to improve the uptake of epithelial cells and viruses (1, 7), to release more microorganisms in vitro (18), and to enhance the molecular detection of Chlamydia trachomatis and Neisseria gonorrhoeae (6). Despite these apparent advantages, a recent study that compared flocked and rayon swabs after a 24-h broth culture enrichment did not show any difference in the rate detection of S. aureus nasal carriers (8). To further assess the utility of flocked swabs, we conducted a study similar to that mentioned just above, with the exception that no enrichment step was performed.Ninety healthy health care workers from the University Hospital of Saint-Etienne, France, were included in the study from March to April 2010. Each volunteer was sampled 7 times with flocked and rayon swabs during a 5-week period (2 volunteers missed the seventh sampling). The study received the approval of our regional ethical research committee (Comité de Protection des Personnes Sud-Est I).A total of 628 nylon swabs (regular flocked swabs, reference 552C; Copan, Brescia, Italy) and 628 rayon swabs (standard swabs with Amies agar gel, reference 108C; Copan) were used by a unique trained fellow by following a predefined protocol (5). Swabs were wetted in normal saline, introduced into the anterior nostril, and rotated 5 times. During each sampling episode, one nostril was randomly sampled with a flocked swab, and the other was sampled with a rayon swab. Each swab was immediately placed into 1 ml of phosphate-buffered saline before being Vortex mixed for 10 s, and 50 μl of this solution was plated onto a chromogenic medium (BBL CHROMagar Staph aureus; Becton Dickinson). After 24 h and 48 h of growth at 37°C, plates were read, and pink colonies were plated onto blood agar. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS; Bruker Daltonics, Bremen, Germany) was used for bacterial identification (9). After 48 h of growth, the number of pink colonies present on the chromogenic medium was determined, according to standard counting procedure, and expressed in CFU/ml. SPSS software (version 16.0, Chicago, IL) was used for statistical analyses. Nonparametric tests were used for mean comparisons. P values below the 5% level were considered statistically significant.A total of 35 of the 90 volunteers (38.9%) were positive for S. aureus in at least one of the nasal samples during the 5-week period. The nasal carriage rate of S. aureus was homogenous throughout the study, with an average of 28.1% ± 1.7% for the 7 sampling episodes. The main results are summarized in Table ​Table1.1. Among the 35 S. aureus nasal carriers, 25 carriers were detected by both of the swabs, 9 by the flocked swab only, and 1 by the rayon swab only (P of 0.026 by the McNemar chi-square test). Assuming that any positive culture result was indicative of positive S. aureus carriage, the sensitivity for the detection of S. aureus nasal carriers was 97.1% (95% confidence interval [CI], 93.6 to 100) with nylon flocked swabs and 74.3% (95% CI, 63.8 to 84.8) with standard rayon swabs. Among the 1,256 swabs taken during 628 samplings episodes, 300 swabs yielded positive cultures of S. aureus, including 160 flocked swabs (53.3%) and 140 rayon swabs (46.7%). The difference was statistically significant (P of 0.01 by the McNemar chi-square test). When considering only the positive samples, the mean loads for S. aureus were 3.41 × 10⁶ CFU/ml with flocked swabs and 4.53 × 10⁵ CFU/ml with rayon swabs (Fig. ​(Fig.11).Open in a separate windowFIG. 1.Bacterial loads of Staphylococcus aureus, shown as the number of log₁₀ CFU/ml, according to the type of swab. The Wilcoxon test was used for statistical comparison.

TABLE 1.

Screening of Staphylococcus aureus nasal carriage, according to the type of swab

Comparison of the Copan WASPLab incorporating the BioRad expert system against the SIRscan 2000 automatic for routine antimicrobial disc diffusion susceptibility testing

ObjectivesThis study investigated the agreement at the categorical level between the Copan WASPLab incorporating the BioRad expert system against the SIRscan 2000 automatic for antimicrobial disc diffusion susceptibility testing.MethodsThe 338 clinical strains (67 Pseudomonas aeruginosa, 19 methicillin-resistant Staphylococcus aureus, 75 methicillin-sensitive S. aureus and 177 Enterobacterales isolates) analysed in this study were non-duplicate isolates obtained from consecutive clinical samples referred to the clinical bacteriology laboratory at Geneva University Hospitals between June and August 2019. For the WASPLab the inoculum suspension was prepared in strict accordance with the manufacturer's instruction (Copan WASP srl, Brescia, Italy) by adding 2 mL of the 0.5 McFarland primary suspension used for the SIRscan analysis into a sterile tube filled with 4 mL of sterile saline (1:3 dilution). The inoculum (2 × 30 μL loop/spreader) was spread over the entire surface of Mueller–Hinton agar plates according to the AST streaking pattern defined by Copan. The antibiotic discs were dispensed by the WASP and inoculated media were loaded on conveyors for transfer to the automatic incubators. The plates were incubated for 16 h, and several digital images were acquired. Inhibition zone diameters were automatically read by the WASPLab and were adjusted manually whenever necessary. For the SIRscan 2000 automatic, the antimicrobial disc diffusion susceptibility testing was performed according to the EUCAST guidelines. The gradient strip method was used to resolve discrepancies.ResultsThe overall categorical agreement between the compared methods reached 99.1% (797/804; 95% CI 98.2%–99.6%), 99.5% (1029/1034; 95% CI 98.9%–99.8%), and 98.8% (2798/2832; 95% CI 98.3%–99.1%) for P. aeruginosa, S. aureus and the Enterobacterales, respectively.ConclusionsWASPLab incorporating the BioRad expert system provides a fully automated solution for antimicrobial disc diffusion susceptibility testing with equal or better accuracy than other available phenotypic methods.     

Comparison of Rates of Positivity for Bordetella pertussis by Real-Time PCR between Specimens Collected with Rayon Swabs on Aluminum Wire Shaft in Amies Gel with Charcoal and Specimens Collected with Flocked Swabs in Universal Viral Transport Medium during an Epidemic

A comparison of real-time PCR positivity rates for Bordetella pertussis between specimens collected with rayon swabs on an aluminum wire shaft in Amies gel with charcoal and those collected with flocked swabs in universal viral transport medium during an epidemic revealed that their performances were comparable.