BackgroundDiseases from Staphylococcus aureus are a major problem in Indian hospitals and recent studies point to infiltration of community associated methicillin resistant S. aureus (CA-MRSA) into hospitals. Although CA-MRSA are genetically different from nosocomial MRSA, the distinction between the two groups is blurring as CA-MRSA are showing multidrug resistance and are endemic in many hospitals. Our survey of samples collected from Indian hospitals between 2004 and 2006 had shown mainly hospital associated methicillin resistant Staphylococcus aureus (HA-MRSA) carrying staphylococcal cassette chromosome mec (SCCmec) type III and IIIA. But S. aureus isolates collected from 2007 onwards from community and hospital settings in India have shown SCCmec type IV and V cassettes while several variations of type IV SCCmec cassettes from IVa to IVj have been found in other parts of the world. In the present study, we have collected nasal swabs from rural and urban healthy carriers and pus, blood etc from in patients from hospitals to study the distribution of SCCmec elements and sequence types (STs) in the community and hospital environment. We performed molecular characterization of all the isolates to determine their lineage and microarray of select isolates from each sequence type to analyze their toxins, virulence and immune-evasion factors.ResultsMolecular analyses of 68 S. aureus isolates from in and around Bengaluru and three other Indian cities have been carried out. The chosen isolates fall into fifteen STs with all major clonal complexes (CC) present along with some minor ones. The dominant MRSA clones are ST22 and ST772 among healthy carriers and patients. We are reporting three novel clones, two methicillin sensitive S. aureus (MSSA) isolates belonging to ST291 (related to ST398 which is live stock associated), and two MRSA clones, ST1208 (CC8), and ST672 as emerging clones in this study for the first time. Sixty nine percent of isolates carry Panton- Valentine Leucocidin genes (PVL) along with many other toxins. There is more diversity of STs among methicillin sensitive S. aureus than resistant ones. Microarray analysis of isolates belonging to different STs gives an insight into major toxins, virulence factors, adhesion and immune evasion factors present among the isolates in various parts of India.ConclusionsS. aureus isolates reported in this study belong to a highly diverse group of STs and CC and we are reporting several new STs which have not been reported earlier along with factors influencing virulence and host pathogen interactions.

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Methicillin-resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen in India, and up to 70% methicillin resistance has been reported from hospitals in various parts of India. Hospitals use phenotyping for the most part, and molecular genotyping is not done. Here we report on the genotyping of 82 single-patient isolates from two hospitals in Bangalore, South India, for the first time. Most of the strains possessed type III or IIIA staphylococcal cassette chromosome (SCCmec) cassettes, and we did not detect strains with type I, IA, or II cassettes. Most isolates also contained the type III cassette chromosome recombinase (ccr) AB region. Multilocus sequence typing (MLST) and staphylococcal protein A (spa) typing of a selected number of isolates have been carried out. Although most isolates that were chosen for MLST and spa typing had the same patterns, they were quite diverse in their pulsed-field gel electrophoresis (PFGE) patterns. PFGE, MLST, and spa typing of the Indian strains revealed that they are related to the previously described Hungarian and Brazilian clones.Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen causing pyogenic, disseminated, and toxinmediated infections (7,18,20). MRSA bacteremia is associated with significantly higher mortality than is known for methicillin-susceptible S. aureus bacteremia (6). Genotyping data from large international studies have shown that a few clones of MRSA are responsible for the spread of the disease in various parts of the world (4,8,18). Methicillin resistance among S. aureus isolates has reached phenomenal proportions in Indian hospitals, with some cities reporting that up to 70% of the strains are resistant to methicillin (2). About 40 to 50% of S. aureus strains isolated from the burn and trauma wards in hospitals in and around Bangalore, India, are resistant (13). For the present study, clinical isolates have been collected from two major hospitals in the city of Bangalore. Many of these MRSA strains are multidrug resistant, and they are characterized only phenotypically at present. The discriminatory power of most of the phenotypic methods is restricted and ambiguous (10, 21). Molecular typing methods have in the last few years paved the way for sophisticated techniques to track the source and transmission route of bacterial pathogens in hospital outbreaks and have also helped in establishing epidemiological investigations comparing strains across continents (1, 4, 23). Pulsed-field gel electrophoresis (PFGE) has been shown to be highly discriminatory in analyzing hospital outbreaks and tracking genetic changes which occur in a relatively short time, while multilocus sequence typing (MLST) is more suitable for studying long-term genetic variations (5,8,16,24). The aim of this study was to characterize the Indian isolates by PFGE, MLST, and spa typing techniques, which would aid in controlling hospital outbreaks, epidemiological studies, and comparison with international strains. MATERIALS AND METHODS Hospitals.St. Joh...

This study provides what we believe to be the first report of the presence of EMRSA-15 and its variants isolated from nasal swabs from 13 healthy and diseased individuals in India. The majority of the isolates belonged to staphylococcal cassette chromosome mec (SCCmec) type IV and spa type t852, whilst four isolates were non-typable and heterotypic for the presence of the mecA gene. All non-typable isolates were positive for the orfX gene by PCR and belonged to spa types t005 and t2986. They may have variant SCCmec cassettes indicating genetic changes occurring in the Indian EMRSA-15. All isolates were positive for Panton-Valentine leukocidin and toxic shock syndrome toxin, which is a cause for concern. In addition to soft-tissue infections, the EMRSA-15 isolates from patients were also responsible for meningitis and brain abscesses, which is quite rare.

PurposeThe purpose of this study was to perform molecular characterization of Staphylococcus aureus isolates causing a variety of eye infections from two major eye care hospitals in India.MethodsTwenty-four isolates from Aravind Eye Hospital, Madurai, India, and nine isolates from LV Prasad Eye Institute, Bhubaneswar, India, representing severe to nonsevere eye infections like microbial keratitis to lacrimal sac abscess, were characterized. Staphylococcal cassette chromosome mec typing, multilocus sequence typing, accessory gene regulator typing, staphylococcal protein A typing, and pulsed field gel electrophoresis were used, along with determination of the presence of Panton–Valentine leucocidin toxin and endotoxin gene cluster among each sequence type.ResultsThe majority of eye infections, both severe and nonsevere, were caused by sequence type (ST)772, positive for the Panton–Valentine leucocidin gene, and carrying methicillin-resistant staphylococcal cassette chromosome mec type V cassette (22/33, 67%). Some of the other sequence types that caused severe eye infections were ST1 (9%), 5 (3%), 72 (6%), 88 (3%), 121 (3%), and 672 (3%). This is the first report of the presence of ST1 and 88 in India.ConclusionAlthough the number of isolates included in this study was small, most of the eye infections were caused by community-associated S. aureus where patients had no history of hospitalization or treatment in the past year. In the case of six severe infections, patients were admitted for surgeries and there is probability of hospital infection. In addition, only methicillin-resistant S. aureus isolates carrying staphylococcal cassette chromosome mec type V were detected. Epidemic methicillin-resistant Staphylococcus aureus 15 (ST22) is a major ST found in health care as well as community settings in non-eye infections in India, but only one methicillin-sensitive S. aureus isolate belonging to ST22 was detected. Predominantly ST772, along with a few other STs, caused the 33 eye infections studied.