Investigating the toxin profiles and clinically relevant antibiotic resistance genes among Staphylococcus aureus isolates using multiplex-PCR assay in Tehran, Iran

Gene Reports ◽  
2020 ◽  
Vol 19 ◽  
pp. 100660
Author(s):  
Hamidreza Houri ◽  
Maryam Samadpanah ◽  
Zahra Tayebi ◽  
Reza Norouzzadeh ◽  
Ebadallah Shiri Malekabad ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Multiplex Pcr ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

scholarly journals Molecular detection of antibiotic resistance genes in multidrug-resistant Staphylococcus aureus isolates from dog dental plaque

2019 ◽  
Vol 22 (4) ◽  
pp. 419-427
Author(s):  
S. Nouri Gharajalar ◽  
M. Onsori
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Multiplex Pcr ◽  
Resistance Genes ◽  
Dental Plaque ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

Multidrug resistant Staphylococcus aureus strains are a major health care problem both in humans and animals. In this work we described three multiplex PCR assays for detection of clinically relevant antibiotic resistance genes in S. aureus isolated from dog dental plaques. Thirty dental plaque samples were collected; then cultural, biochemical and molecular tests performed for isolation and identification of S. aureus from samples. The antibiotic susceptibility of the isolates were checked by Kirby Bauer disc diffusion method and the prevalence of antibiotic resistance genes determined using multiplex PCR assay. As a result S. aureus was isolated from 18 dog plaque samples. Fifteen of these isolates were resistant to penicillin. The mecA gene was more prevalent than blaZ among penicillin-resistant bacteria. Ten of the isolates were resistant to tetracycline. The percentage of tetM was higher than tetK among them. Also, 10 of the isolates were resistant to cefazolin among them bla TEM detected in higher rate than blaSHV and blaOXA-1. Hence multiplex PCR assay is a suitable method for detection of antibiotic resistance patterns of S. aureus isolates.

Multiplex PCR detection of the antibiotic resistance genes in Staphylococcus aureus strains isolated from auricular infections

2008 ◽  
Vol 53 (4) ◽  
pp. 357-362 ◽  
Author(s):  
T. Zmantar ◽  
K. Chaieb ◽  
F. Ben Abdallah ◽  
A. Ben Kahla-Nakbi ◽  
A. Ben Hassen ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Multiplex Pcr ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Pcr Detection

scholarly journals Erythromycin and Clindamycin Resistance in Group B Streptococcal Clinical Isolates

2006 ◽  
Vol 50 (5) ◽  
pp. 1875-1877 ◽  
Author(s):  
Scott E. Gygax ◽  
Jessica A. Schuyler ◽  
Lauren E. Kimmel ◽  
Jason P. Trama ◽  
Eli Mordechai ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Susceptibility Testing ◽  
Antibiotic Resistance Genes ◽  
Clinical Isolates ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Disk Diffusion Assay ◽  
Group B ◽  
Diffusion Assay

ABSTRACT Erythromycin (EM) and clindamycin (CM) susceptibility testing was performed on 222 clinical isolates of group B Streptococcus. A multiplex PCR assay was used to detect the ermB, ermTR, and mefA/E antibiotic resistance genes. These results were compared to the phenotypes as determined by the standard EM/CM double disk diffusion assay.

scholarly journals Correlation between the Resistance Genotype Determined by Multiplex PCR Assays and the Antibiotic Susceptibility Patterns of Staphylococcus aureus andStaphylococcus epidermidis

2000 ◽  
Vol 44 (2) ◽  
pp. 231-238 ◽  
Author(s):  
Francis Martineau ◽  
François J. Picard ◽  
Nicolas Lansac ◽  
Christian Ménard ◽  
Paul H. Roy ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Multiplex Pcr ◽  
Resistance Gene ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Disk Diffusion ◽  
Erythromycin Resistance ◽  
Oxacillin Resistance ◽  
Pcr Assays

ABSTRACT Clinical isolates of Staphylococcus aureus (a total of 206) and S. epidermidis (a total of 188) from various countries were tested with multiplex PCR assays to detect clinically relevant antibiotic resistance genes associated with staphylococci. The targeted genes are implicated in resistance to oxacillin (mecA), gentamicin [aac(6′)-aph(2")], and erythromycin (ermA, ermB, ermC, andmsrA). We found a nearly perfect correlation between genotypic and phenotypic analysis for most of these 394 strains, showing the following correlations: 98% for oxacillin resistance, 100% for gentamicin resistance, and 98.5% for erythromycin resistance. The discrepant results were (i) eight strains found to be positive by PCR for mecA or ermC but susceptible to the corresponding antibiotic based on disk diffusion and (ii) six strains of S. aureus found to be negative by PCR for mecA or for the four erythromycin resistance genes targeted but resistant to the corresponding antibiotic. In order to demonstrate in vitro that the eight susceptible strains harboring the resistance gene may become resistant, we subcultured the susceptible strains on media with increasing gradients of the antibiotic. We were able to select cells demonstrating a resistant phenotype for all of these eight strains carrying the resistance gene based on disk diffusion and MIC determinations. The four oxacillin-resistant strains negative for mecA were PCR positive for blaZand had the phenotype of β-lactamase hyperproducers, which could explain their borderline oxacillin resistance phenotype. The erythromycin resistance for the two strains found to be negative by PCR is probably associated with a novel mechanism. This study reiterates the usefulness of DNA-based assays for the detection of antibiotic resistance genes associated with staphylococcal infections.

scholarly journals Novel Multiplex PCR Assay for Detection of Chlorhexidine-Quaternary Ammonium, Mupirocin, and Methicillin Resistance Genes, with Simultaneous Discrimination of Staphylococcus aureus from Coagulase-Negative Staphylococci

2017 ◽  
Vol 55 (6) ◽  
pp. 1857-1864 ◽  
Author(s):  
Jo-Ann McClure ◽  
Johanna Zaal DeLongchamp ◽  
John M. Conly ◽  
Kunyan Zhang
Keyword(s):  
Staphylococcus Aureus ◽  
Multiplex Pcr ◽  
Resistance Genes ◽  
Quaternary Ammonium ◽  
Methicillin Resistance ◽  
Rrna Gene ◽  
Pcr Assay ◽  
Coagulase Negative Staphylococci ◽  
Content Type ◽  
Multiplex Pcr Assay

ABSTRACTMethicillin-resistantStaphylococcus aureus(MRSA) is a clinically significant pathogen that is resistant to a wide variety of antibiotics and responsible for a large number of nosocomial infections worldwide. The Agency for Healthcare Research and Quality and the Centers for Disease Control and Prevention recently recommended the adoption of universal mupirocin-chlorhexidine decolonization of all admitted intensive care unit patients rather than MRSA screening with targeted treatments, which raises a serious concern about the selection of resistance to mupirocin and chlorhexidine in strains of staphylococci. Thus, a simple, rapid, and reliable approach is paramount in monitoring the prevalence of resistance to these agents. We developed a simple multiplex PCR assay capable of screeningStaphylococcusisolates for the presence of antiseptic resistance genes for chlorhexidine and quaternary ammonium compounds, as well as mupirocin and methicillin resistance genes, while simultaneously discriminatingS. aureusfrom coagulase-negative staphylococci (CoNS). The assay incorporates 7 PCR targets, including theStaphylococcus16S rRNA gene (specifically detectingStaphylococcusspp.),nuc(distinguishingS. aureusfrom CoNS),mecA(distinguishing MRSA from methicillin-susceptibleS. aureus),mupAandmupB(identifying high-level mupirocin resistance), andqacandsmr(identifying chlorhexidine and quaternary ammonium resistance). Our assay demonstrated 100% sensitivity, specificity, and accuracy in a total of 23 variant antiseptic- and/or antibiotic-resistant control strains. Further validation of our assay using 378 randomly selected and previously well-characterized local clinical isolates confirmed its feasibility and practicality. This may prove to be a useful tool for multidrug-resistantStaphylococcusmonitoring in clinical laboratories, particularly in the wake of increased chlorhexidine and mupirocin treatments.

scholarly journals Molecular characteristics and virulence gene profiles of Staphylococcus aureus isolates in Hainan, China

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Xuehan Li ◽  
Tao Huang ◽  
Kai Xu ◽  
Chenglin Li ◽  
Yirong Li
Keyword(s):  
Staphylococcus Aureus ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Virulence Genes ◽  
Mainland China ◽  
Geographical Location ◽  
Antibiotic Resistance Genes ◽  
Virulence Gene ◽  
Molecular Characteristics ◽  
Staphylococcal Protein A

Abstract Background There have been no reports regarding the molecular characteristics, virulence features, and antibiotic resistance profiles of Staphylococcus aureus (S. aureus) from Hainan, the southernmost province of China. Methods Two hundred twenty-seven S. aureus isolates, consisting of 76 methicillin-resistant S. aureus (MRSA) and 151 methicillin-susceptible S. aureus (MSSA), were collected in 2013–2014 and 2018–2019 in Hainan, and investigated for their molecular characteristics, virulence genes, antibiotic resistance profiles and main antibiotic resistance genes. Results Forty sequence types (STs) including three new STs (ST5489, ST5492 and ST5493), and 79 Staphylococcal protein A (spa) types were identified based on multilocus sequence typing (MLST) and spa typing, respectively. ST398 (14.1%, 32/227) was found to be the most prevalent, and the prevalence of ST398-MSSA increased significantly from 2013 to 2014 (5.5%, 5/91) to 2018–2019 (18.4%, 25/136). Seventy-six MRSA isolates were subject to staphylococcus chromosomal cassette mec (SCCmec) typing. SCCmec-IVa was the predominant SCCmec type, and specifically, ST45-SCCmec IVa, an infrequent type in mainland China, was predominant in S. aureus from Hainan. The antibiotic resistance profiles and antibiotic resistance genes of S. aureus show distinctive features in Hainan. The resistant rates of the MRSA isolates to a variety of antibiotics were significantly higher than those of the MSSA isolates. The predominant erythromycin and tetracycline resistance genes were ermC (90.1%, 100/111) and tetK (91.8%, 78/85), respectively. Eleven virulence genes, including the Panton-Valentine leukocidin (pvl) and eta, were determined, and the frequency of eta and pvl were found to be 57.3 and 47.6%. Such high prevalence has never been seen in mainland China before. Conclusion S. aureus isolates in Hainan have unique molecular characteristics, virulence gene and antibiotic resistance profiles, and main antibiotic resistance genes which may be associated with the special geographical location of Hainan and local trends in antibiotic use.

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