Detection of heterogeneous vancomycin-intermediate Staphylococcus aureus: A preliminary report from south India.

Background & objectives: Although there are reports of heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) across the globe, there is a lack of reliable data on hVISA in India. The present study was undertaken to determine the rate of hVISA among the methicillin-resistant Staphylococcus aureus (MRSA) isolates, and to compare the brain heart infusion agar with vancomycin 4 µg/ml (BHIV4) method with population analysis profile-area under the curve (PAP-AUC) method for the detection of hVISA and to study the distribution of mobile genetic element that carries methicillin-resistance gene SCCmec (Staphylococcal cassette chromosome mec) types among these isolates. Methods: BHIV4 and PAP-AUC methods were employed to detect hVISA among 500 clinical isolates of MRSA. SCCmec typing of these isolates was performed by multiplex polymerase chain reaction. The clinical presentation, treatment with vancomycin and outcome was documented for patients with hVISA. Results: The rate of hVISA was 12.4 per cent by PAP-AUC method. Sensitivity, specificity, PPV, NPV and kappa agreement of BHIV4 with PAP-AUC was 58.06, 93.15, 54.55, 94.01 per cent and 0.498, respectively. The isolation of hVISA was significantly (P<0.01) higher in patients admitted to intensive care units and wards than in patients attending the outpatient departments. Only 38 per cent of the patients received vancomycin as therapy. Majority of the hVISA isolates carried SCCmec type V or IV. Interpretation & conclusions: The rate of hVISA isolation in our study was 12.4 per cent. The sensitivity of the BHIV4 screening test was low, and was in moderate agreement with PAP-AUC test. SCCmec type V was the predominant type seen in half of the isolates. More studies need to be done in different parts of the country on a large number of isolates to confirm our findings.

Existence of multiple SCCmec elements in clinical isolates of methicillin-resistant Staphylococcus aureus.

PURPOSE: Methicillin-resistant Staphylococcus aureus (MRSA) can be classified into hospital-acquired MRSA (HA-MRSA) and community-associated MRSA (CA-MRSA), based on the associated epidemiological risk factors and their SCCmec types. We therefore studied the diversity and distribution of SCCmec elements among MRSA isolates in our region and also evaluated SCCmec typing as a tool for the classification of MRSA. METHODOLOGY: Two hundred isolates of MRSA obtained from various clinical specimens were included. The clinical and demographic details of the patients and the epidemiological risk factors for MRSA acquisition were documented. Multiplex PCR was optimized for all the major SCCmec types (I to V). Subtyping of SCCmec type IV (IVb, IVc, IVd, IVh) was carried out by simplex PCR. RESULTS: Based on epidemiological criteria, CA-MRSA constituted 57  % (114/200) of the the test isolates and HA-MRSA made up 43  % (86/200). The predominant SCCmec type found in our study was type III (62%), followed by type V (52.5%) and type I (47.5%), while type II was carried by a single isolate. Of the 200 isolates, 118 carried multiple SCCmec types and 3 were non-typable. CONCLUSION: The existence of multiple SCCmec types in individual MRSA isolates resulted in our inability to categorize many of these isolates as either CA-MRSA or HA-MRSA as defined by the SCCmec type criterion. LIMITATION: The major limitation of the study was that the SCC mec element of MRSA isolates exhibiting multiple types was not sequenced and hence this finding could not be confirmed.

Is there a need to revise the antibiotic concentration in Clinical and Laboratory Standards Institute-Recommended Oxacillin Screen Agar?

INTRODUCTION: In routine diagnostic microbiology laboratories, Clinical and Laboratory Standards Institute (CLSI) recommends the use of cefoxitin disc, in addition to oxacillin screen agar (OSA) of 6 µg/ml for the detection of methicillin-resistant Staphylococcus aureus (MRSA), whereas minimum inhibitory concentration values of oxacillin for S. aureus are ≤2 µg/ml (susceptible) and ≥4 µg/ml (resistant). Hence, the study was carried out to evaluate the ability of screen agar with lower concentrations of oxacillin to identify the isolates of MRSA and to compare this with cefoxitin disc diffusion (CDD). MATERIALS AND METHODS: Six hundred and seventy-six isolates of S. aureus were screened for methicillin resistance by OSA with 2 µg/ml and 4 µg/ml and 6 µg/ml of oxacillin concentration as well as CDD. Polymerase chain reaction for mecA gene was carried out for all isolates which grew on OSA 2, 4 and 6 µg/ml regardless of their cefoxitin susceptibility. Latex agglutination test for penicillin-binding protein 2a was performed for the isolates which grew on OSA 2 and or 4 µg/ml but not on OSA 6 µg/ml. RESULTS: Eight per cent of MRSA isolates was missed by using OSA 6 µg/ml, when compared with other methods. Sensitivities of OSA 2 µg/ml, OSA 6 µg/ml and CDD were found to be 100%, 92.5% and 97.5%, respectively, and specificities for the same were found to be 100%, 100% and 98%, respectively. As per FDA criteria, categorical agreement for OSA 2 µg/ml was found to be 100% in comparison with the reference broth microdilution method. No major and very major discrepancies were documented. CONCLUSION: Similar findings on a larger and more heterogeneous collection of isolates may indicate the need to revise the concentration of OSA to 2 µg/ml for the detection of MRSA.

Prevalence and genetic mechanisms of antimicrobial resistance in Staphylococcus species: A multicentre report of the indian council of medical research antimicrobial resistance surveillance network.

PURPOSE: Routine surveillance of antimicrobial resistance (AMR) is an essential component of measures aimed to tackle the growing threat of resistant microbes in public health. This study presents a 1-year multicentre report on AMR in Staphylococcus species as part of Indian Council of Medical Research-AMR surveillance network. MATERIALS AND METHODS: Staphylococcus species was routinely collected in the nodal and regional centres of the network and antimicrobial susceptibility testing was performed against a panel of antimicrobials. Minimum inhibitory concentration (MIC) values of vancomycin (VAN), daptomycin, tigecycline and linezolid (LNZ) against selected methicillin-resistant Staphylococcus aureus(MRSA) isolates were determined by E-test and MIC creep, if any, was determined. Resistant genotypes were determined by polymerase chain reaction for those isolates showing phenotypic resistance. RESULTS: The prevalence of MRSA was found to be range from moderate (21%) to high (45%) among the centres with an overall prevalence of 37.3%. High prevalence of resistance was observed with commonly used antimicrobials such as ciprofloxacin and erythromycin in all the centres. Resistance to LNZ was not encountered except for a single case. Full-blown resistance to VAN in S. aureus was not observed; however, a few VAN-intermediate S. aureus isolates were documented. The most common species of coagulase negative staphylococci (CoNS) identified was Staphylococcus haemolyticus and Staphylococcus epidermidis. Resistance among CoNS was relatively higher than S. aureus. Most phenotypically resistant organisms possessed the corresponding resistance genes. CONCLUSION: There were localised differences in the prevalence of resistance between the centres. The efficacy of the anti-MRSA antimicrobials was very high; however, almost all these antimicrobials showed evidence of creeping MIC.

Can Panton Valentine Leukocidin Gene And Clindamycin Susceptibility Serve As Predictors of Community Origin of MRSA From Skin and Soft Tissue Infections?

INTRODUCTION: Community associated Methicillin-resistant Staphylococcus aureus (CA-MRSA) strains have begun to replace Hospital Associated MRSA (HA-MRSA) strains in hospital settings all over the world. With the epidemiological distinctions between these strains beginning to become ill-defined, the categorisation of a strain as CA-MRSA or HA-MRSA is dependent on molecular methods to detect the presence of SCCmec (Staphylococcal Cassette Chromosome mec) elements. However other markers like the presence of Panton Valentine Leukocidin toxin (pvl) genes or Clindamycin susceptibility may also be associated with community origin of MRSA. AIM: To determine the prevalence of CA-MRSA among MRSA strains isolated from skin and soft tissue infections and to evaluate the usefulness of Panton Valentine Leukocidin and Clindamycin susceptibility as markers of community origin of MRSA. MATERIALS AND METHODS: One hundred isolates of MRSA from skin and soft tissue were studied for the presence of SCCmec IV and V genes and Panton valentine leukocidin gene by Polymerase chain reaction. Inducible clindamycin resistance was screened for using the D-test. STATISTICAL ANALYSIS USED: Fischer's exact test. A p-value <0.05 was considered significant. RESULTS: Eighteen out of 100 MRSA strains were found to be CA-MRSA based on presence of SCCmecV. The proportion of Panton Valentine Leukocidin gene carriage among CA- MRSA as compared to HA-MRSA was found to be statistically significant (p<0.0001). Among the CA-MRSA strains, 94.4% were found to be susceptible to Clindamycin as against only 13.4% of the HA-MRSA strains (p<0.0001). The odds of an MRSA strain being CA-MRSA if it was both Clindamycin susceptible and PVL gene positive was calculated to be 68.25 (p<0.0001). CONCLUSION: Both Clindamycin susceptibility and pvl gene carriage were found to be independent predictors of community origin of MRSA, but taken together the association was highly significant.