Influence of Broth Enrichment as well as Storage and Transport Time on the Sensitivity of MRSA Surveillance in the Tropics.

Direct growth on blood and screening agar for methicillin-resistant Staphylococcus aureus (MRSA) at a tropical surveillance site was compared with broth enrichment and subsequent growth on selective MRSA agar after international sample transport. In Madagascar, 1548 swabs from an MRSA surveillance study were assessed for growth on Columbia blood agar enriched with 5% sheep blood and MRSA screening agar at the surveillance site with subsequent cold storage of the samples and shipment to Germany. In Germany, 1541 shipped samples were analyzed by non-selective broth enrichment with subsequent culture on MRSA selective agar. A total of 28 MRSA isolates were detected. Of these, 20 strains were isolated from direct culture on blood and MRSA screening agars at the surveillance site, 24 MRSA strains were isolated using the broth enrichment method in Germany, and 16 MRSA strains were identified by both approaches. In spite of the observed die-off of individual strains due to long-term storage and transport, broth enrichment with subsequent screening on MRSA selective agar after international sample shipment led to comparable sensitivity of MRSA detection like streaking on blood and MRSA agar at the tropical surveillance site.

Prevalence of nasal colonisation by methicillin-sensitive and methicillin-resistant Staphylococcus aureus among healthcare workers and students in Madagascar.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) clones pose a significant threat to hospitalised patients because the bacteria can be transmitted by asymptomatic carriers within healthcare facilities. To date, nothing is known about the prevalence of S. aureus and MRSA among healthcare workers in Madagascar. The objective of our study was to examine the prevalence and clonal epidemiology of nasal S. aureus and MRSA among healthcare workers and non-medical University students in Antananarivo, Madagascar. METHODS: This cross sectional study screened nasal swabs taken from students and healthcare workers for S. aureus. Multiplex PCR was performed to identify S. aureus-specific (nuc), MRSA-specific mecA and mecC genes, Panton-Valentine leukocidin (PVL) (lukF-PV), and toxic shock syndrome toxin-1 (TSST-1) specific genes in methicillin-sensitive S. aureus (MSSA) and MRSA isolates. Staphylococcus protein A gene (spa) typing was performed for all confirmed MRSA isolates. The frequency distribution of nasal S. aureus and MRSA of healthcare workers and non-medical students was compared using Pearson's χ(2) test. RESULTS: Of 1548 nasal swabs tested, 171 (11 %) were positive for S. aureus; 20 (1.3 %) of these isolates were identified as MRSA. S. aureus was detected in 91 of 863 healthcare workers (10.4 %) and in 80 (11.8 %) of 685 students; however, 14 (1.5 %) healthcare workers carried MRSA compared with six (0.9 %) students. Nasal carriage of S. aureus and MRSA was more prevalent in women than in men, and 21 (11.7 %) S. aureus isolates were PVL-positive and 36 (21 %) were TSST-1 positive. The mecC gene was not detected in any isolates. Five different spa types were identified, with spa type t186 being the predominant MRSA clone (16/20). CONCLUSION: The results of the present study reveal a low frequency of S. aureus and MRSA nasal carriage in both students and healthcare workers from Antananarivo, Madagascar. The predominant MRSA clone (t186) was previously described in hospitalised patients in Madagascar.

Screening agars for MRSA: evaluation of a stepwise diagnostic approach with two different selective agars for the screening for methicillin-resistant Staphylococcus aureus (MRSA).

BACKGROUND: Colonization with methicillin-resistant Staphylococcus aureus (MRSA) poses a hygiene risk that does not spare field hospitals or military medical field camps during military deployments. Diagnostic options for unambiguously identifying MRSA isolates are usually scarce in military environments. In this study, we assessed the stepwise application of two different selective agars for the specific identification of MRSA in screening analyses. METHODS: Nasal swabs from 1541 volunteers were subjected to thioglycollate broth enrichment and subsequently screened on CHROMagar MRSA selective agar for the identification of MRSA. The MRSA identity of suspicious-looking colonies was confirmed afterwards or excluded by another selective agar, chromID MRSA. All isolates from the selective agars with MRSA-specific colony morphology were identified by biochemical methods and mass spectrometry. RESULTS: The initial CHROMagar MRSA screening identified suspicious colonies in 36 out of 1541 samples. A total of 25 of these 36 isolates showed MRSA-like growth on chromID agar. Out of these 25 isolates, 24 were confirmed as MRSA, while one isolate was identified as Staphylococcus kloosii. From the 11 strains that did not show suspicious growth on chromID agar, 3 were methicillin-sensitive Staphylococcus aureus (MSSA, with one instance of co-colonization with Corynebacterium spp.), 2 were confirmed as MRSA (with 1 instance of co-colonization with MSSA), 2 were lost during passaging and could not be re-cultured, one could not be identified by the applied approaches, and the remaining 3 strains were identified as Staphylococcus saprophyticus, Staphylococcus hominis (co-colonized with Macrococcus caseolyticus) and Staphylococcus cohnii, respectively. CONCLUSIONS: The application of the selective agar CHROMagar MRSA alone proved to be too non-specific to allow for a reliable diagnosis of the presence of MRSA. The combined use of two selective agars in a stepwise approach reduced this non-specificity with an acceptably low loss of sensitivity. Accordingly, such a stepwise screening approach might be an option for resource-restricted military medical field camps.

Identification of nasal colonization with ß-lactamase-producing Enterobacteriaceae in patients, health care workers and students in Madagascar.

This study assesses the nasal occurrence of ß-lactamase-producing Enterobacteriaceae both in patients in a hospital department of infectious diseases at admission and in healthy Madagascan students and health care workers. Nasal swabs from 681 students, 824 health care workers, and 169 patients were obtained in Antananarivo, Madagascar, and transferred to Germany. Screening for ß-lactamase (ESBL, ampC) producing Enterobacteriaceae was performed by cultural and molecular approaches, comprising Brilliance ESBL agar, E-testing, ABCD-testing, and commercial hyplex ESBL and SuperBug ID PCR. Regarding ESBL-positive strains and strains with resistance against at least three out of the four tested bactericidal antibiotic drugs, 0.3% (five out of 1541) of the students and health care workers group showed nasal colonization, whereas colonization was observed in 7.1% (12 out of 169) of the hospitalized patients at admission. No appreciably reduced detection rates after sample storage and intercontinental transport were observed. A considerable proportion of nasal colonization with cephalosporin-resistant Enterobacteriaceae was demonstrated in Madagascan hospital patients at admission, posing a risk of developing future endogenous infections. The nasal colonization of healthy individuals was negligible. Good storage and transport stability of Enterobacteriaceae will allow for future studies even in areas difficult to access.