Efficient differentiation of Corynebacterium striatum , Corynebacterium amycolatum and Corynebacterium xerosis clinical isolates by multiplex PCR using novel species-specific primers

Santos, Carolina S.A.B.; Ramos, Juliana Nunes; Vieira, Verônica Viana; Pinheiro, Carina S.; Meyer, Roberto; Alcantara-Neves, Neuza Maria; Ramos, Rommel Thiago Jucá; Silva, Artur M. S.; Hirata, Raphael; Felicori, Liza; Puig, Carlos Ruiz de Alegría; Navas, Jesús; Azevedo, Vasco; Mattos‐Guaraldi, Ana Luíza; Pacheco, Luis Gustavo Carvalho

doi:10.1016/j.mimet.2017.09.002

Cited by 9 publications

(7 citation statements)

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“…Several studies have shown that C. amycolatum infections are often misidentified by culturing and subsequent phenotypic analysis of the isolates, making it difficult to implement appropriate therapeutic interventions ( Funke et al, 1996 ; Zinkernagel et al, 1996 ; Wauters et al, 1998 ; Soltan Mohammadi et al, 2013 ). In this sense, it is essential to define better phenotypic and genetic markers that could improve the identification of pathogenic nonlipophilic members of the genus Corynebacterium , including C. amycolatum ( Santos et al, 2017 , 2018 ). C. amycolatum can be clearly distinguished from C. xerosis and C. imitans by means of MALDI-TOF mass spectrometry using the MALDI Biotyper system ( Alibi et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

“…which are difficult-to-differentiate from C. amycolatum in phenotypic tests, particularly C. xerosis ( Santos et al, 2018 ). Besides, through comparative genomics we were able to identify specific target genes that can render reliable identification of C. striatum , C. amycolatum and C. xerosis clinical isolates, by multiplex PCR ( Santos et al, 2017 ). More recently, different studies have been demonstrating the added value of whole-genome analyses to improve species circumscription in the genus Corynebacterium , including the study by Dover and collaborators ( Dover et al, 2021 ) which proposes a new phylogenomic-based classification of the genus Corynebacterium , based on previous studies ( Huson and Bryant, 2006 ), encompassing 19 phylogenetic groups; C. amycolatum belongs to the newly proposed group M, that also includes isolates of C. xerosis and C. freneyi ( Dover et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

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Pan-genomic analysis of Corynebacterium amycolatum gives insights into molecular mechanisms underpinning the transition to a pathogenic phenotype

et al. 2022

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Corynebacterium amycolatum is a nonlipophilic coryneform which is increasingly being recognized as a relevant human and animal pathogen showing multidrug resistance to commonly used antibiotics. However, little is known about the molecular mechanisms involved in transition from colonization to the MDR invasive phenotype in clinical isolates. In this study, we performed a comprehensive pan-genomic analysis of C. amycolatum, including 26 isolates from different countries. We obtained the novel genome sequences of 8 of them, which are multidrug resistant clinical isolates from Spain and Tunisia. They were analyzed together with other 18 complete or draft C. amycolatum genomes retrieved from GenBank. The species C. amycolatum presented an open pan-genome (α = 0.854905), with 3,280 gene families, being 1,690 (51.52%) in the core genome, 1,121 related to accessory genes (34.17%), and 469 related to unique genes (14.29%). Although some classic corynebacterial virulence factors are absent in the species C. amycolatum, we did identify genes associated with immune evasion, toxin, and antiphagocytosis among the predicted putative virulence factors. Additionally, we found genomic evidence for extensive acquisition of antimicrobial resistance genes through genomic islands.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Pan-genomic analysis of Corynebacterium amycolatum gives insights into molecular mechanisms underpinning the transition to a pathogenic phenotype

et al. 2022

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“…Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) has been applied to perform accurate species-level identification of Corynebacterium spp. clinical isolates, but this technology is not yet fully accessible to clinical microbiology laboratories in resource-limited settings [17]. Also, MALDI-TOF requires pure cultures as starting material, which precludes rapid diagnosis.…”

Section: Introductionmentioning

confidence: 99%

Direct detection of Corynebacterium striatum, Corynebacterium propinquum, and Corynebacterium simulans in sputum samples by high-resolution melt curve analysis

Qiu

Hou

et al. 2021

BMC Infect Dis

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Background Pulmonary infections caused by non-diphtheriae corynebacteria are increasing. However, rapid identification of Corynebacterium species poses a challenge due to the low genetic variation within the genus. Methods Three reference strains and 99 clinical isolates were used in this study. A qPCR followed by high-resolution melting (HRM) targeting ssrA was performed to simultaneously identify C. striatum, C. propinquum and C. simulans. To further evaluate this assay’s performance, 88 clinical sputum samples were tested by HRM and the detection results were compared with those of the traditional culture method and multiple cross-displacement amplification (MCDA) assay. Results The melting curve produced by a pair of universal primers generated species-specific HRM curve profiles and could distinguish the three target species from other related bacteria. The limit of detection of HRM assay for DNA from the three purified Corynebacterium species was 100 fg. Compared with the culture method, HRM detected 22 additional positive specimens, representing a 23.9% relative increase in detection rate. The HRM assay had 98.4% (95% confidence interval [CI], 90.5–99.9%) sensitivity and 100% (95% CI, 82.8–100%) specificity. Additionally, 95.5% concordance between HRM and MCDA (κ = 0.89 [95% CI, 0.79–0.99]) was noted. Conclusions The HRM assay was a simple, rapid, sensitive, and specific diagnostic tool for detecting C. striatum, C. propinquum, and C. simulans, with the potential to contribute to early diagnosis, epidemiological surveillance, and rapid response to outbreak.

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“…Within recent years there has been a considerable increase in the number of literature data reporting non-diphtherial corynebacteria (also known as diphtheroids) as the causative agents of opportunistic and nosocomial infections in humans (De Miguel-Martinez et al 1996;Yoon et al 2011;Bernard 2012b;Nhan et al 2012;Qin et al 2017;Santos et al 2017;Kang et al 2018). These bacteria are members of the microbiota of the skin and mucous membranes (Qin et al 2017;Santos et al 2017;Kang et al 2018). The potential association between coryneforms and clinical infections can be attributed to immunosuppression, severe underlying medical disorders or invasive procedures (Nhan et al 2012;Cacopardo et al 2013;Kimura et al 2017;Qin et al 2017).…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cytokine Levels in the In Vitro Response of T Cells to Planktonic and Biofilm Corynebacterium amycolatum

Olender

Bogut

Magryś

et al. 2019

Polish Journal of Microbiology

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Unravelling of the interplay between the immune system and non-diphtheria corynebacteria would contribute to understanding their increasing role as medically important microorganisms. We aimed at the analysis of pro- (TNF, IL-1β, IL-6, IL-8, and IL-12p70) and anti-inflammatory (IL-10) cytokines produced by Jurkat T cells in response to planktonic and biofilm Corynebacterium amycolatum. Two reference strains: C. amycolatum ATCC 700207 (R-CA), Staphylococcus aureus ATCC 25923 (R-SA), and ten clinical strains of C. amycolatum (C-CA) were used in the study. Jurkat T cells were stimulated in vitro by the planktonic-conditioned medium (PCM) and biofilm-conditioned medium (BCM) derived from the relevant cultures of the strains tested. The cytokine concentrations were determined in the cell culture supernatants using the flow cytometry. The levels of the cytokines analyzed were lower after stimulation with the BCM when compared to the PCM derived from the cultures of C-CA; statistical significance (p < 0.05) was observed for IL-1β, IL-12 p70, and IL-10. Similarly, planktonic R-CA and R-SA stimulated a higher cytokine production than their biofilm counterparts. The highest levels of pro-inflammatory IL-8, IL-1β, and IL-12p70 were observed after stimulation with planktonic R-SA whereas the strongest stimulation of anti-inflammatory IL-10 was noted for the BCM derived from the mixed culture of both reference species. Our results are indicative of weaker immunostimulatory properties of the biofilm C. amycolatum compared to its planktonic form. It may play a role in the persistence of biofilm-related infections. The extent of the cytokine response can be dependent on the inherent virulence of the infecting microorganism.

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Efficient differentiation of Corynebacterium striatum , Corynebacterium amycolatum and Corynebacterium xerosis clinical isolates by multiplex PCR using novel species-specific primers

Cited by 9 publications

References 15 publications

Pan-genomic analysis of Corynebacterium amycolatum gives insights into molecular mechanisms underpinning the transition to a pathogenic phenotype

Pan-genomic analysis of Corynebacterium amycolatum gives insights into molecular mechanisms underpinning the transition to a pathogenic phenotype

Direct detection of Corynebacterium striatum, Corynebacterium propinquum, and Corynebacterium simulans in sputum samples by high-resolution melt curve analysis

Cytokine Levels in the In Vitro Response of T Cells to Planktonic and Biofilm Corynebacterium amycolatum

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