High Resolution Melting Analysis for Rapid Mutation Screening in Gyrase and Topoisomerase IV Genes in Quinolone-Resistant<i>Salmonella enterica</i>

Ngoi, Soo Tein; Thong, Kwai Lin

doi:10.1155/2014/718084

Cited by 9 publications

(6 citation statements)

References 28 publications

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“… 47 , 50 Moreover, most of the isolates had an additional Asp87Asn mutations in gyr A (88.9%) and Ser80Ile mutation in par C (77.8%), which is associated with high level of ciprofloxacin resistance. 51–53 Similar mutations were also reported by Al-Marzooq et al, 54 Ngoi et al, 55 Zeng et al, 56 and Guimarães et al 57 On the other hand, the identified mutations are different from those observed by Azargun et al 58 Resistance of Enterobacteriaceae to quinolones have been reported with alterations in genes other than the gyr A and par C. Lindbäck, Rahman, Jalal, Wretlind 59 and others have related mutation in gyr B and par E to resistance to quinolones. 60 No other mutations were observed in par C, however, other investigators have reported a Glu84Val substitution as the most common mutation in the par C gene.…”

Section: Discussionsupporting

confidence: 81%

Detection of gyrA and parC Mutations and Prevalence of Plasmid-Mediated Quinolone Resistance Genes in Klebsiella pneumoniae

Kareem¹,

Al-Kadmy²,

Kazaal³

et al. 2021

IDR

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Background and Aim Recently, the extensive use of quinolones led to increased resistance to these antimicrobial agents, with different rates according to the organism and the geographical region. The aim of this study was to detect the resistance rate of Klebsiella pneumoniae Iraqi isolates toward quinolone antimicrobial agents, to determine genetic mutations in gyrA and parC , to screen for efflux-pump activity, and to screen the presence of plasmid-mediated quinolone resistance (PMQR) genes. Methods Forty-three K. pneumoniae isolates were confirmed phenotypically and genotypically by Vitek 2 system and species specific primers by PCR using the targeting rpo gene followed by sequencing. Antibiotic susceptibility test was carried out using disc diffusion method. Quinolone resistant isolates were subjected to ciprofloxacin MIC testing, and cartwheel method to screen for efflux pump activity. The presence of the plasmid mediated quinolone resistance genes qepA, qnrB, qnrS , and aac(6)Ib was tested by PCR. Sequencing of gyr A and par C was performed. Results We observed a high rate of resistance to ceftriaxone, gentamicin ciprofloxacin, and levofloxacin. Low rate of resistance was detected against amikacin and azithromycin. Ciprofloxacin MIC results revealed that 96.1% of the isolates had MICs >256 µg/mL, 83.4% had MICs >512 µg/mL while 34.6% had MIC >1024 µg/mL. Testing of isolates against ciprofloxacin mixed with EtBr at various concentrations resulted in decreased resistant. Sequencing results showed that Ser83Leu was the most common mutation in gyr A that was observed in all quinolone resistant isolates, followed by Asp87Asn. Ser80Ile mutation in par C was observed in 77.7% of the tested isolates. The prevalence of PMQR genes was 92.5% aac (6)-Ib , 51.8% qnr B, 40.7% qep A, and 37% qnr S. Conclusion Quinolone resistance is common in K. pneumoniae isolates in Baghdad. The frequent mutation in gyr A and par C, and the presence of PMQR genes is alarming.

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Section: Discussionsupporting

confidence: 81%

Detection of gyrA and parC Mutations and Prevalence of Plasmid-Mediated Quinolone Resistance Genes in Klebsiella pneumoniae

Kareem¹,

Al-Kadmy²,

Kazaal³

et al. 2021

IDR

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“…Mutations within QRDR of parC and parE genes were rarely detected and reported among Salmonella isolates [25] [31] [32]. The finding of double mutations in parE gene in this study are remarkable, and may be associated with higher level of FQ resistance [30] [31]. In addition to it, parE gene mutations were identified in three S. Enteritidis isolates that were having gyrA gene muta-tions too.…”

Section: Discussionmentioning

confidence: 51%

Molecular Detection of Mutations within the Quinolone Resistance-Determining Regions in Non Typhoidal Salmonella Isolates from Malaysia

Mohd-Nor¹,

Abu-Bakar²,

Mohd-Isa³

et al. 2017

Health

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Introduction: The efficacy of chemotherapy in bacteraemia caused by nontyphoidal Salmonella (NTS) is compromised by antibiotic resistance. Objective: This study was undertaken to describe the mechanism of resistance among clinical NTS isolates. Materials & Methodology: Thirty of NTS were isolated from blood (n = 19), stool (n = 10) and bronchioalveolar lavage (BAL; n = 1) respectively. These isolates were tested for susceptibility testing by disc diffusion method against ampicillin, gentamicin, tetracycline, co-trimoxazole, nalidixic acid, ciprofloxacin and ceftriaxone. Epsilometer tests (E-test) for nalidixic acid and ciprofloxacin were performed for nalidixic acid resistant isolates by disc diffusion method. DNA sequencing was carried out on six of the nalidixic acid resistant Salmonella Enteritidis isolates to identify mutations within quinolones resistance determining regions (QRDR) of gyrA, gyrB, parC and parE genes. Results: Resistance rates of NTS isolates from blood, stool, and BAL were respectively 37%, 20% and 0% for ampicillin, 79%, 40% and 0% for tetracycline, 32%, 40% and 0% for co-trimoxazole, 37%, 10% and 100% for nalidixic acid. Eight isolates were resistant to nalidixic acid and had exhibited reduced susceptibility towards ciprofloxacin by E-test. Mutation

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“…Previous research reported that the underlying mechanism for ciprofloxacin resistances may be caused by specific mutations in genes encoding DNA gyrase and topoisomerase IV that decrease quinolone sensitivity by weakening the interactions between quinolones and bacterial enzymes [ 27 ]. With the availability of WGS data and ResFinder’ sister database, PointFinder, two mutations were detected in gyrA (S83F and D87G) in strain SS20, confirming their associations with ciprofloxacin resistance [ 28 ].…”

Section: Discussionmentioning

confidence: 99%

Prevalence of IncFIB Plasmids Found among Salmonella enterica Serovar Schwarzengrund Isolates from Animal Sources in Taiwan Using Whole-Genome Sequencing

Chen

et al. 2021

Pathogens

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Salmonella enterica serovar Schwarzengrund is one of the most frequently isolated Salmonella serotypes responsible for human and poultry infections in Taiwan, and it has raised public health concerns. To better facilitate the understanding of transmission patterns and the dynamics of epidemics, sharing molecular data on pathogen profiles is urgently needed. The objectives of the current study were to determine and establish baseline data of S. enterica serovar Schwarzengrund isolates from 23 epidemiologically unrelated sources from year 2000 to 2018 and examine their phenotypic and genotypic characteristics. Genomic DNA of the Salmonella isolates was extracted and subjected to whole-genome sequencing using an Illumina platform. Results showed that all selected isolates exhibited multidrug resistance, and six of those were resistant to ciprofloxacin phenotypically. Genotypically, these isolates carried genes resistant to aminoglycoside (100%), phenicol (91.3%), β-lactams (69.5%), folate pathway antagonist (100%), tetracycline (82.6%), and fluoroquinolone (4.3%). Moreover, these isolates harbor integrons with five different gene cassettes identified for the first time, which are associated with resistance to trimethoprim, streptomycin, tetracycline, sulfonamide, chloramphenicol, and gentamicin. Furthermore, prevalence of IncFIB plasmid was found among studied isolates, which may increase its ability to colonize the chicken cecum and cause extra-intestinal disease. Salmonella pathogenicity islands SPI-1 to SPI-5, SPI-13, and SPI-14, as well as C63PI locus, were also detected in all isolates. This study demonstrated that a considerable high antimicrobial resistance with high virulence levels of Salmonella were found from animal sources. Sharing data on these pathogen profiles can not only help increase the reproducibility and accessibility of genomic analysis but can also support surveillance and epidemiological investigations for salmonellosis in the region.

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High Resolution Melting Analysis for Rapid Mutation Screening in Gyrase and Topoisomerase IV Genes in Quinolone-ResistantSalmonella enterica

Cited by 9 publications

References 28 publications

Detection of gyrA and parC Mutations and Prevalence of Plasmid-Mediated Quinolone Resistance Genes in Klebsiella pneumoniae

Detection of gyrA and parC Mutations and Prevalence of Plasmid-Mediated Quinolone Resistance Genes in Klebsiella pneumoniae

Molecular Detection of Mutations within the Quinolone Resistance-Determining Regions in Non Typhoidal Salmonella Isolates from Malaysia

Prevalence of IncFIB Plasmids Found among Salmonella enterica Serovar Schwarzengrund Isolates from Animal Sources in Taiwan Using Whole-Genome Sequencing

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