Prevalence and genotypic relatedness of methicillin resistant Staphylococcus aureus in a tertiary care hospital

Fomda, BA; Thokar, Manzoor Ahmad; Bashir, Gulnaz; Khan, Ajaz Kariem; Kour, Avneet; Zahoor, Danish; Ray, Pallab

doi:10.4103/0022-3859.143964

Cited by 5 publications

(4 citation statements)

References 10 publications

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“…Out of these, 350 blood cultures were excluded from the study as they did not meet the inclusion criteria. (1), S. hominis (5), S. hemolyticus (6), S. capitis (1).…”

Section: Resultsmentioning

confidence: 99%

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Direct Detection of Methicillin Resistant Staphylococci: Comparison of Phenotypic Methods with Multiplex PCR and Direct Susceptibility Testing

Bashir

Nissar

Wani

et al. 2020

JAMB

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Bloodstream infections are augean diseases characterized by a high morbidity and mortality, related with the lag in administration of the first adequate anti-infectious agent. Clinical and epidemiological data guide the clinicians towards empirical anti-infectious treatments whose effectualness remains questionable especially in the present day milieu of multidrug-resistant organisms. Early microbiological evidence of the causative agent is hierophantic of antimicrobial stewardship. We evaluated three rapid methods for the direct identification of S. aureus from 720 positive blood cultures in comparison to Vitek 2 automated microbial identification system. Early distinction S. aureus from CoNS was attempted with Direct Tube Coagulase. Multiplex PCR assay was used to separate MRSA from MSSA by the presence of mecA gene. Direct antibiotic susceptibility (DST) from positive blood culture bottles was performed in an endeavour to reduce time and compared to the routine Vitek 2 compact antimicrobial susceptibility. For Direct tube coagulase at 4 hrs of incubation sensitivity was 79.6% while specificity was 100% however, on overnight incubation sensitivity increased to 97.9% with 100% specificity. The positive predictive value and negative predictive values were 100% and 96.9% respectively when compared to Vitek 2 identification of staphylococcal species. By DST Categorical agreement of 95% was seen for coagulase negative staphylococci microorganism-antibiotic combinations. Categorical agreement of 89.7% was seen for S. aureus microorganism-antibiotic combinations. Direct multiplex PCR testing did not misidentify any S. aureus isolate compared to Vitek 2. In 12 methicillin resistant CoNS isolates mecA gene was not detected by PCR. In total 13(3.5%) strains of staphylococci identified by DST as methicillin resistant were not identified by PCR analysis. Each of the tests has positive qualities and all may have a place in a Gram positive cocci algorithm for testing blood cultures depending on the laboratory setting, workload volume and staffing. However, rapid detection methods are a pressing priority.

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“…Out of these, 350 blood cultures were excluded from the study as they did not meet the inclusion criteria. (1), S. hominis (5), S. hemolyticus (6), S. capitis (1).…”

Section: Resultsmentioning

confidence: 99%

“…In north India, the prevalence of MRSA was 46 % where MRSA isolates were found to be more resistant to other antibiotics than Methicillin sensitive Staphylococcus aureus (MSSA). There was a prevalence of 39.48% MRSA reported from Sheri-Kashmir Institute of Medical Sciences in 2014 [6].…”

Section: Introductionmentioning

confidence: 99%

Direct Detection of Methicillin Resistant Staphylococci: Comparison of Phenotypic Methods with Multiplex PCR and Direct Susceptibility Testing

Bashir

Nissar

Wani

et al. 2020

JAMB

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“…The methicillin resistance determinant gene mecA and the heat-stable nuclease gene nucA were amplified from S. aureus isolates as 533 bp and 270 bp DNA fragments, respectively, using primers and amplification protocols described previously [ 56 ]. Antimicrobial resistance genes encoding aminoglycoside resistance ( aac(6′)-aph(2″), ant(4′)-Ia, aph(3′)-IIIa, aac(6′)-Ie-aph(2′)-Ia, str , and sat4 ) and genes conferring resistance to beta-lactams ( mecA and blaZ ), macrolides ( mphC, ermC, and ermB ), tetracycline ( tetK and tetM ), and chloramphenicol ( catpC221 and catpC223 ) were amplified by using PCR primers and conditions described in previous publications [ 57 , 58 , 59 , 60 , 61 ].…”

Section: Methodsmentioning

confidence: 99%

Genotypic Characterization of Clinical Isolates of Staphylococcus aureus from Pakistan

et al. 2021

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In this study, we compared pulsed-field gel electrophoretic (PFGE), multilocus sequence typing (MLST), Staphylococcal cassette chromosome mec (SCCmec), spa typing, and virulence gene profiles of 19 Panton–Valentine leucocidin (PVL)-positive, multidrug-, and methicillin-resistant clinical Staphylococcus aureus (MRSA) isolates obtained from a hospital intensive care unit in Pakistan. The isolates exhibited 10 pulsotypes, contained eight adhesin genes (bbp, clfA, clfB, cna, fnbA, fnbB, map-eap, and spa), 10 toxin genes (hla, hlb, hld, hlg, pvl, sed, see, seg, seh, and tst), and two other virulence genes (cfb, v8) that were commonly present in all isolates. The spa-typing indicated seven known spa types (t030, t064, t138, t314, t987, t1509, and t5414) and three novel spa types. MLST analysis indicated eight ST types (ST8, ST15, ST30, ST239, ST291, ST503, ST772, and ST1413). All isolates belonged to the agr group 1. Most of the isolates possessed SCCmec type III, but some isolates had it in combination with types SCCmec IV and V. The presence of multidrug-resistant MRSA isolates in Pakistan indicates poor hygienic conditions, overuse of antibiotics, and a lack of rational antibiotic therapy that have led to the evolution and development of hypervirulent MRSA clones. The study warrants development of a robust epidemiological screening program and adoption of effective measures to stop their spread in hospitals and the community.

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“…Methicillin resistance in Staphylococci is conferred by mecA gene that produces altered PBP2a. Detection of mecA gene remains the gold standard for identification of methicillin resistance; however it does not confirm the species S. aureus [18]. There is no consensus on the genomic target that could be used to confirm the S. aureus.…”

Section: Introductionmentioning

confidence: 99%

A Simple Sample Processing Protocol and Multiplex PCR for Direct Detection of MRSA from Uncultured Clinical Samples—A Pilot Study

Chikkala¹,

Ch²,

Divyakolu³

et al. 2019

AID

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Phenotypic tests have limited discrimatory power to identify closely related members of genus Staphylococcus and particularly for identification of S. aureus. 157 isolates of S. aureus obtained from different clinical specimens were included in our study. To present a demonstration of our method's sensitivity and ability to correctly detect S. aureus from uncultured clinical specimen, 30 known S. aureus positive but leftover uncultured clinical specimens from clinical microbiology laboratory were processed by our protocol and analyzed. All the 30 clinical specimens were confirmed as S. aureus among which 26 specimen were identified as MRSA and the remaining 4 as MSSA. These 30 clinical specimens used in the study showed 100% correlation with coagulase test and Cefoxitin disc diffusion method. Though commercial molecular diagnostic kits are available for detecting MRSA from swabs, this is probably the first time that multiplex PCR is being demonstrated directly on a variety of uncultured clinical specimens.

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Prevalence and genotypic relatedness of methicillin resistant Staphylococcus aureus in a tertiary care hospital

Cited by 5 publications

References 10 publications

Direct Detection of Methicillin Resistant Staphylococci: Comparison of Phenotypic Methods with Multiplex PCR and Direct Susceptibility Testing

Direct Detection of Methicillin Resistant Staphylococci: Comparison of Phenotypic Methods with Multiplex PCR and Direct Susceptibility Testing

Genotypic Characterization of Clinical Isolates of Staphylococcus aureus from Pakistan

A Simple Sample Processing Protocol and Multiplex PCR for Direct Detection of MRSA from Uncultured Clinical Samples—A Pilot Study

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