Updated Multiplex PCR for Detection of All Six Plasmid-Mediated
            <i>qnr</i>
            Gene Families

Kraychete, Gabriela Bergiante; Botelho, Larissa Alvarenga Batista; Campana, Eloiza Helena; Picão, Renata C.; Bonelli, Raquel Regina

doi:10.1128/aac.01447-16

Cited by 32 publications

(13 citation statements)

References 8 publications

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“…The DNA extracts were used for molecular screening of qnr genes. PCR amplification for detecting qnrA , qnrB , qnrC , qnrD , qnrS , and qnrVC was conducted using primers and conditions as previously described [ 20 , 21 ] ( Table S1 ). Product amplification was performed in a Bio-Rad CFX96 Touch Real-Time PCR Detection System (Bio-Rad, Feldkirchen, Germany).…”

Section: Methodsmentioning

confidence: 99%

Phenotypic and Genotypic Properties of Fluoroquinolone-Resistant, qnr-Carrying Escherichia coli Isolated from the German Food Chain in 2017

et al. 2021

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Fluoroquinolones are the highest priority, critically important antimicrobial agents. Resistance development can occur via different mechanisms, with plasmid-mediated quinolone resistance (PMQR) being prevalent in the livestock and food area. Especially, qnr genes, commonly located on mobile genetic elements, are major drivers for the spread of resistance determinants against fluoroquinolones. We investigated the prevalence and characteristics of qnr-positive Escherichia (E.) coli obtained from different monitoring programs in Germany in 2017. Furthermore, we aimed to evaluate commonalities of qnr-carrying plasmids in E. coli. We found qnr to be broadly spread over different livestock and food matrices, and to be present in various sequence types. The qnr-positive isolates were predominantly detected within selectively isolated ESBL (extended spectrum beta-lactamase)-producing E. coli, leading to a frequent association with other resistance genes, especially cephalosporin determinants. Furthermore, we found that qnr correlates with the presence of genes involved in resistance development against quaternary ammonium compounds (qac). The detection of additional point mutations in many isolates within the chromosomal QRDR region led to even higher MIC values against fluoroquinolones for the investigated E. coli. All of these attributes should be carefully taken into account in the risk assessment of qnr-carrying E. coli from livestock and food.

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Section: Methodsmentioning

confidence: 99%

Phenotypic and Genotypic Properties of Fluoroquinolone-Resistant, qnr-Carrying Escherichia coli Isolated from the German Food Chain in 2017

et al. 2021

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“…Screening of E. coli isolates for ARG types was conducted using PCR for each isolate depending on the antibiotic resistance phenotype. The ARGs we examined were (1) carbapenemase genes bla NDM , bla IMP bla KPC , bla VIM , bla OXA , bla AIM , bla BIC , bla DIM , bla GIM , bla SIM , and bla SPM including bla NDM and bla CTX-M subtyping (Poirel et al, 2011), (2) β-lactamase genes bla SHV , bla TEM , and bla CTX-M (Casella et al, 2018), (3) plasmidmediated AmpC β-lactamase genes bla MOX , bla CMY , bla LAT , bla DHA , bla ACC , bla MIR , bla ACT , and bla FOX (Perez-Perez and Hanson, 2002), (4) colistin resistance genes mcr-1-8 (Rebelo et al, 2018;Yang et al, 2018;Carroll et al, 2019), and (5) plasmid-mediated quinolone resistance (PMQR) genes qnrA, qnrB and qnrS including whether the gyrA and parC genes in the quinolone resistance determining region (QRDR) were mutated (Komp et al, 2003;Kraychete et al, 2016;Onseedaeng and Ratthawongjirakul, 2016). PCR primers used for screening are shown in Supplementary Table S1.…”

Section: Arg Detectionmentioning

confidence: 99%

Increasing Prevalence of ESBL-Producing Multidrug Resistance Escherichia coli From Diseased Pets in Beijing, China From 2012 to 2017

et al. 2019

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We investigated antimicrobial resistance trends and characteristics of ESBL-producing Escherichia coli isolates from pets and whether this correlates with antibiotic usage in the clinic. Clinical samples containing E. coli from diseased cats and dogs were screened for antibiotic sensitivity and associated genotypic features. We identified 127 E. coli isolates from 1886 samples from dogs (n = 1565) and cats (n = 321) with the majority from urinary tract infections (n = 108, 85%). High rates of resistance were observed for β-lactams and fluoroquinolones and resistance to > 3 antibiotic classes (MDR) increased from 67% in 2012 to 75% in 2017 (P < 0.0001). This was especially true for strains resistant to 6-9 antibiotics that increased from 26.67 to 60.71%. Increased rates in β-lactam use for clinical treatment accompanied these increasing resistance rates. Accordingly, the most frequently encountered subtypes were bla CTX-M (n = 44, 34.65%), bla CTX-M-65 (n = 19) and bla CTX-M-15 (n = 18) and qnrB (n = 119, 93.70%). The bla CTX-M-isolates possessed 36 unique pulsed field electrophoretic types (PFGEs) and 28 different sequence types (STs) in ST405 (7, 15.9%), ST131 (3, 6.8%), ST73, ST101, ST372, and ST827 (2, 4.5% each) were the most prevalent. This data demonstrated a high level of diversity for the bla CTX-M-positive E. coli isolates. Additionally, bla NDM-5 was detected in three isolates (n = 3, 2.36%), comprised of two ST101 and one ST405 isolates, and mcr-1 was also observed in three colistin-resistant E. coli with three different STs (ST6316, ST405, and ST46). Our study demonstrates an increasing trend in MDR and ESBL-producing E. coli and this correlated with β-lactam antibiotic usage for treatment of these animals. This data indicates that there is significant risk for the spread of resistant bacteria from pets to humans and antibiotic use for pets should be more strictly regulated.

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“…The DNA extracted from soil samples was subjected to PCR amplification of quinolone-resistance genes (qnrA, qnrB and qnrS) in separate reactions. PCR was performed in 25-µl reactions containing 1× PCR buffer, 2 U of Taq DNA polymerase (Promega), 200 µM each deoxynucleotide triphosphate (dNTP), 5 pmol of each forward and reverse primer (listed in Kraychete et al, 2016), 1.5 mM MgCl 2 , and 2 µl template DNA (20-40 ng). PCR conditions were: 10 min at 95 • C; 25 cycles of amplification consisting of 45 s at 95 • C, 45 s at 58 • C and 15 s at 72 • C; and then 3 min at 72 • C (described in detail by Kraychete et al, 2016).…”

Section: Detection Of Quinolone-resistance (Qnr) Genesmentioning

confidence: 99%

“…PCR was performed in 25-µl reactions containing 1× PCR buffer, 2 U of Taq DNA polymerase (Promega), 200 µM each deoxynucleotide triphosphate (dNTP), 5 pmol of each forward and reverse primer (listed in Kraychete et al, 2016), 1.5 mM MgCl 2 , and 2 µl template DNA (20-40 ng). PCR conditions were: 10 min at 95 • C; 25 cycles of amplification consisting of 45 s at 95 • C, 45 s at 58 • C and 15 s at 72 • C; and then 3 min at 72 • C (described in detail by Kraychete et al, 2016). DNA fragments were analyzed by electrophoresis in a 1.4% agarose gel at 80 V for 1 h in 1× TBE buffer followed by staining with ethidium bromide.…”

Section: Detection Of Quinolone-resistance (Qnr) Genesmentioning

confidence: 99%

Response of the Bacterial Communities Associated With Maize Rhizosphere to Poultry Litter as an Organomineral Fertilizer

Vollú

Cotta

Jurelevicius

et al. 2018

Front. Environ. Sci.

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Maize is an important food source worldwide and is of considerable industrial importance. Low maize yields are mostly due to low soil fertility, so expensive mineral fertilizers are often used to offset the lack of nutrients. Poultry litter (PL) is one of the most valuable and phosphorous-rich animal wastes. However, PL usually contains veterinary antibiotic residues, particularly fluoroquinolones (FQs), which may alter soil microorganism diversity and resistance patterns. In this study, we aimed to understand the impact of applying mineral (triple superphosphate-STP) or organomineral (STP with PL and reactive Bayovar phosphate with PL) fertilizers (130 or 260 kg/ha of total P 2 O 5) on the structure and composition of the soil bacteriome and on phosphate-mineralizing bacteria associated with the maize rhizosphere. Maize plants were sampled at 60 and 90 days after sowing and a clear rhizosphere effect was observed in all samples. No specific groups of bacterial genera predominated (>3% relative abundance) according to the different fertilizer treatments and most of the genera were shared among samples. Multivariate analyses of 16S rRNA sequences revealed clear clustering based on sampling time and distinct separation from bulk soil samples. Abundances of phosphate-mineralizing bacteria varied depending on the sampling time. We observed a positive effect on phytase activity under the 260 kg STP with PL treatment. Although the FQ enrofloxacin and its main metabolite ciprofloxacin were detected in PL, their concentrations in fertilized soils were below quantification thresholds. Quinolone resistance genes were not detected in the maize rhizosphere or bulk soil. Together, these results suggest that the rhizosphere effect, plant age and applied amounts of fertilizer are more influential on bacterial communities than the type of fertilizer used. Thus, application of PL as an organomineral fertilizer does not appear to have extensive impacts on the bacterial diversity of maize rhizosphere, so it could be an excellent option for enhancing maize production.

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Updated Multiplex PCR for Detection of All Six Plasmid-Mediated qnr Gene Families

Cited by 32 publications

References 8 publications

Phenotypic and Genotypic Properties of Fluoroquinolone-Resistant, qnr-Carrying Escherichia coli Isolated from the German Food Chain in 2017

Phenotypic and Genotypic Properties of Fluoroquinolone-Resistant, qnr-Carrying Escherichia coli Isolated from the German Food Chain in 2017

Increasing Prevalence of ESBL-Producing Multidrug Resistance Escherichia coli From Diseased Pets in Beijing, China From 2012 to 2017

Response of the Bacterial Communities Associated With Maize Rhizosphere to Poultry Litter as an Organomineral Fertilizer

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