LAURO VIEIRA PERDIGAO NETO

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LIM/49 - Laboratório de Protozoologia, Hospital das Clínicas, Faculdade de Medicina

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Autor e Coautores FMUSP-HC Normalizados: ROBERTA CRISTINA RUEDAS MARTINS ×

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  • article 21 Citação(ões) na Scopus
    Multidrug-resistant Stenotrophomonas maltophilia: Description of new MLST profiles and resistance and virulence genes using whole-genome sequencing
    (2018) RIZEK, Camila Fonseca; JONAS, Daniel; PAEZ, Jorge Isaac Garcia; ROSA, Juliana Ferraz; PERDIGAO NETO, Lauro Vieira; MARTINS, Roberta Ruedas; MORENO, Luisa Z.; ROSSI JUNIOR, Alfio; LEVIN, Anna S.; COSTA, Silvia Figueiredo
    Objectives: Stenotrophomonas maltophilia is an opportunistic pathogen that has high intrinsic and acquired antimicrobial resistance, with great genetic diversity. The aim of this study was to characterise four S. maltophilia clinical isolates displaying different susceptibility profiles using whole-genome sequencing. Methods: The whole genomes of four clinical isolates of S. maltophilia from three patients were sequenced using Ion Torrent (TM) PGM technology. The isolates presented different susceptibilities to trimethoprim/sulfamethoxazole (SXT) and levofloxacin. Results: Three new multilocus sequence typing (MLST) profiles were identified (ST144, ST172 and ST173), differing in virulence and resistance genes. The ST172 isolate had more genes related to toxins than related to motility or adhesion and had different types of efflux pumps than the other isolates. The SXT-resistant strains belonged to ST172 or ST144 and did not harbour the sul1, sul2 or dfrA resistance genes. Strains I and II, from the same patient and belonging to the same ST but differing in resistance to SXT, had all of the resistance genes searched for in common, except for the SmeABC efflux pump complex genes that were only found in the SXT-resistant strain. All strains, including the strain susceptible to levofloxacin, harboured the qnrB gene, which may question the importance of this gene in determining levofloxacin resistance in S. maltophilia. Conclusion: Here we describe three new MLST profiles. Resistance to SXT in these strains appears to be associated with efflux pumps.
  • article 33 Citação(ões) na Scopus
    Ceftazidime-Avibactam as Salvage Therapy for Infections Caused by Enterobacteriales Coresistant to Carbapenems and Polymyxins
    (2019) GUIMARAES, Thais; NOUER, Simone A.; MARTINS, Roberta C. R.; V, Lauro Perdigao Neto; MARTINS, Willames M. B. S.; BARBOSA, Ana Clara Narciso; FERREIRA, Adriana L. P.; COSTA, Silvia F.; GALES, Ana C.
    In this article, we report a case series of patients with infections caused by Enterobacteriales coresistant to carbapenems and polymyxins who were treated with ceftazidime/avibactam (CAZ-AVI) salvage therapy on a compassionate-use protocol. We enrolled 29 adult patients in 3 centers that had an infection due to a resistant microorganism and for whom the treatments available were considered ineffective, treated them with CAZ-AVI, and assessed clinical and microbiological cure at the end of treatment and all-cause mortality at 14 days and 30 days. The antimicrobial susceptibility profile was determined using broth microdilution, and total genomic DNA was sequenced. Twelve (41.4%) patients had bacteremia, and 48.3% (14/29) of the infections were treated with combination therapy. All strains were producers of KPC-2 and were susceptible to CAZ-AVI (MIC90, 1 mu g/ml). Clinical success was high (24/29 [82.7%; 95% confidence interval, 64.2 to 94.2%]), even for the bacteremic cases (75%). The 14-day and 30-day mortality rates were 9/29 (31%) and 15/29 (51.7%), respectively. The 14-day mortality rate for pneumonia was the same as that for bloodstream infections (33.3%) and although not significant, we found that patients with renal impairment that received adjusted doses of CAZ-AVI had high mortality (4/9 (44%); P = 0.22). We concluded that CAZ-AVI is an option for the treatment of severe infections due to difficult-to-treat drug-resistant Enterobacteriales.
  • article 6 Citação(ões) na Scopus
    Prevalence of Clostridioides difficile associated diarrhea in hospitalized patients in five Brazilian centers: A multicenter, prospective study
    (2020) GIRAO, Evelyne Santana; TAVARES, Bruno de Melo; SANTOS, Sania Alves dos; GAMARRA, Gessica Lorena; RIZEK, Camila; MARTINS, Roberta Cristina; NETO, Lauro Vieira Perdigao; DIOGO, Constancia; ORSI, Tatiana D'Annibale; ESPINOZA, Evelyn Patricia Sanchez; MORALES, Hugo Manuel Paz; NOGUEIRA, Keite da Silva; MAESTRI, Adriane Ceshin; BOSZCZOWSKI, Icaro; PIASTRELLI, Filipe; COSTA, Cecilia Leite; COSTA, Daniely Viana; MACIEL, Geovania; ROMAO, Janete; GUIMARAES, Thais; BRITO, Gerly Anne de Castro; COSTA, Silvia Figueiredo
    Epidemiological data on CD infection (CDI) in Latin American are scarce. CDI prevalence and strains characterization were prospectively evaluated in 5 Brazilian hospitals from different regions. Prevalence rates of CDI were 15%, ranging from 0 to 37%. ST42 was the most common Sequence Type and hyper virulent strains were not identified.
  • article 10 Citação(ões) na Scopus
    Comparison of methods for the detection of in vitro synergy in multidrug-resistant gram-negative bacteria
    (2020) GAUDERETO, Juliana Januario; PERDIGAO NETO, Lauro Vieira; LEITE, Gleice Cristina; SANCHEZ, Evelyn; MARTINS, Roberta Cristina Ruedas; PRADO, Gladys Villas Boas; ROSSI, Flavia; GUIMARAES, Thais; LEVIN, Anna Sara; COSTA, Silvia Figueiredo
    Background The use of combined antibiotic therapy has become an option for infections caused by multidrug-resistant (MDR) bacteria. The time-kill (TK) assay is considered the gold standard method for the evaluation of in vitro synergy, but it is a time-consuming and expensive method. The purpose of this study was to evaluate two methods for testing in vitro antimicrobial combinations: the disk diffusion method through disk approximation (DA) and the agar gradient diffusion method via the MIC:MIC ratio. The TK assay was included as the gold standard. MDR Gram-negative clinical isolates (n = 62; 28 Pseudomonas aeruginosa, 20 Acinetobacter baumannii, and 14 Serratia marcescens) were submitted to TK, DA, and MIC:MIC ratio synergy methods. Results Overall, the agreement between the DA and TK assays ranged from 20 to 93%. The isolates of A. baumannii showed variable results of synergism according to TK, and the calculated agreement was statistically significant in this species against fosfomycin with meropenem including colistin-resistant isolates. The MIC:MIC ratiometric agreed from 35 to 71% with TK assays. The kappa test showed good agreement for the combination of colistin with amikacin (K = 0.58; P = 0.04) among the colistin-resistant A. baumannii isolates. Conclusions The DA and MIC:MIC ratiometric methods are easier to perform and might be a more viable tool for clinical microbiology laboratories.
  • article 12 Citação(ões) na Scopus
    Alternative drugs against multiresistant Gram-negative bacteria
    (2020) PERDIGAO NETO, Lauro Vieira; OLIVEIRA, Maura Salaroli; ORSI, Tatiana D'Annibale; PRADO, Gladys Villas Boas do; MARTINS, Roberta Cristina Ruedas; LEITE, Gleice Cristina; MARCHI, Ana Paula; LIRA, Esther Sant'Ana de; CORTES, Marina Farrel; ESPINOZA, Evelyn Patricia Sanchez; CARRILHO, Claudia Maria Dantas de Maio; BOSZCZOWSKI, Icaro; GUIMARAES, Thais; COSTA, Silvia Figueiredo; LEVIN, Anna S.
    Objectives: Enterobacterales and other non-fermenting Gram-negative bacteria have become a threat worldwide owing to the frequency of multidrug resistance in these pathogens. On the other hand, efficacious therapeutic options are quickly diminishing. The aims of this study were to describe the susceptibility of 50 multiresistant Gram-negative bacteria, mostly pan-resistant, against old and less-used antimicrobial drugs and to investigate the presence of antimicrobial resistance genes. Methods: A total of 50 genetically distinct isolates were included in this study, including 14 Acinetobacter baumannii (belonging to ST79, ST317, ST835 and ST836), 1 Pseudomonas aeruginosa (ST245), 8 Serratia marcescens and 27 Klebsiella pneumoniae (belonging to STII, ST340, ST258, ST16, ST23, ST25, ST101, ST234, ST437 and ST442). The isolates were submitted to antimicrobial susceptibility testing and whole-genome sequencing to evaluate lineages and resistance genes. Results: Our results showed that some strains harboured carbapenemase genes, e.g. bla(K)(PC-)(2) (28/50; 56%) and bla(OXA-23) (11/50; 22%), and other resistance genes encoding aminoglycoside-modifying enzymes (49/50; 98%). Susceptibility rates to tigecycline (96%) in all species (except P. aeruginosa), to minocycline (100%) and doxycycline (93%) in A. baumannii, to ceftazidime/avibactam in S. marcescens (100%) and K. pneumoniae (96%), and to fosfomycin in S. marcescens (88%) were high. Chloramphenicol and quinolones (6% susceptibility each) did not perform well, making their use in an empirical scenario unlikely. Conclusions: This study involving genetically distinct bacteria showed promising results for tigecycline for all Gram-negative bacteria (except P. aeruginosa), and there was good activity of minocycline against A. baumannii, ceftazidime/avibactam against Enterobacterales, and fosfomycin against S. marcescens. (C) 2020 The Author(s).
  • article 14 Citação(ões) na Scopus
    Colistin-resistant Klebsiella pneumoniae co-harboring KPC and MCR-1 in a Hematopoietic Stem Cell Transplantation Unit
    (2019) HIGASHINO, Hermes Ryoiti; MARCHI, Ana Paula; MARTINS, Roberta Cristina Ruedas; BATISTA, Marjorie Vieira; PERDIGAO NETO, Lauro Vieira; LIMA, Victor Augusto Camarinha de Castro; ROSSI, Flavia; GUIMARAES, Thais; LEVIN, Anna Sara; ROCHA, Vanderson; COSTA, Silvia Figueiredo
  • article 6 Citação(ões) na Scopus
    Genetic and virulence characterization of colistin-resistant and colistin-sensitive A. baumannii clinical isolates
    (2019) LEITE, Gleice Cristina; STABLER, Richard A.; NEVES, Patricia; PERDIGAO NETO, Lauro V.; MARTINS, Roberta C. Ruedas; RIZEK, Camila; ROSSI, Flavia; LEVIN, Anna S.; COSTA, Silvia Figueiredo
    Treatment of infections caused by A. baumannii is becoming a challenge due to the ability to develop multidrug-resistance, virulence, and high mortality. We described the colistin resistance and virulence genes present in sixA. baumannii clinical isolates using WGS, expression by qPCR, and virulence in the Galleria mellonella model. The colistin-resistant isolates were assigned as ST233 and the colistin-susceptible isolates as ST236 and ST407. The colistin-resistant isolates contained mutations within PmrA/PmrB, and the pmrA showed up-regulation in all of them. Only one colistin-resistant isolate indicating virulence in G. mellonella. This particular isolate belonged to a different clone, and it was the only isolate that presented non-synonymous mutations in pmrB. Colistinresistance in A. baumannii isolates seems to be caused by up-regulation of pmrA gene. Only one isolate appeared to be virulent in the G. mellonella model. This finding indicating low virulence in isolates belonging to emerging clones circulating in our hospital.
  • article 17 Citação(ões) na Scopus
    Vancomycin-resistant enterococci isolates colonizing and infecting haematology patients: clonality, and virulence and resistance profile
    (2018) MARCHI, A. P.; PERDIGAO NETO, L. V.; MARTINS, R. C. R.; RIZEK, C. F.; CAMARGO, C. H.; MORENO, L. Z.; MORENO, A. M.; BATISTA, M. V.; BASQUEIRA, M. S.; ROSSI, F.; AMIGO, U.; GUIMARAES, T.; LEVIN, A. S.; COSTA, S. F.
    Background: Vancomycin-resistant enterococci (VRE) are an important agent of colonization and infection in haematology patients. However, the role of virulence on VRE colonization and infection is controversial. Aim: To characterize the lineage, virulence and resistance profile of VRE infection and colonization isolates; as well as their impact on outcome of haematology patients using a regression logistic model. Methods: Eighty-six isolates (80 Enterococcus faecium and six E. faecalis) from 76 patients were evaluated. Polymerase chain reaction for resistance and virulence genes, and pulsed-field gel electrophoresis and whole genome sequencing of the major clusters, were performed. Bivariate and multivariate analyses were carried out to evaluate the role of virulence genes on outcome. Findings: All isolates harboured the vanA gene. Regarding the virulence genes, 96.5% of isolates were positive for esp, 69.8% for gelE and asa1 genes. VRE infection isolates were more virulent than colonization isolates and harboured more often the gelE gene (P = 0.008). Infections caused by VRE carrying asal gene resulted more frequently in death (P = 0.004), but only the predominant clone remained as protector in the multivariate model. The E. faecium strains were assigned to seven STs (ST78, ST412, ST478, ST792, ST896, ST987, ST963) that belonged to CC17. The E. faecalis sequenced belonged to ST9 (CC9). Conclusion: E. faecium was predominant, and infection isolates were more virulent than colonization isolates and harboured more often the gene gelE. Infections caused by VRE carrying the asal gene appeared to be associated with a fatal outcome.
  • article 14 Citação(ões) na Scopus
    Bloodstream Infections caused by Klebsiella pneumoniae and Serratia marcescens isolates co-harboring NDM-1 and KPC-2
    (2021) BES, Taniela; NAGANO, Debora; MARTINS, Roberta; MARCHI, Ana Paula; PERDIGAO-NETO, Lauro; HIGASHINO, Hermes; PRADO, Gladys; GUIMARAES, Thais; LEVIN, Anna S.; COSTA, Silvia
    Carbapenem-resistant Enterobacteriaceae are a worldwide health problem and isolates carrying both bla(KPC-2) and bla(NDM-1) are unusual. Here we describe the microbiological and clinical characteristics of five cases of bloodstream infections (BSI) caused by carbapenem-resistant Klebsiella pneumoniae and Serratia marcescens having both bla(KPC-2) and bla(NDM-1). Of the five blood samples, three are from hematopoietic stem cell transplantation patients, one from a renal transplant patient, and one from a surgical patient. All patients lived in low-income neighbourhoods and had no travel history. Despite antibiotic treatment, four out of five patients died. The phenotypic susceptibility assays showed that meropenem with the addition of either EDTA, phenylboronic acid (PBA), or both, increased the zone of inhibition in comparison to meropenem alone. Molecular tests showed the presence of bla(KPC-2) and bla(NDM-1) genes. K. pneumoniae isolates were assigned to ST258 or ST340 by whole genome sequencing. This case-series showed a high mortality among patients with BSI caused by Enterobacteriae harbouring both carbapenemases. The detection of carbapenemase-producing isolates carrying both bla(KPC-2) and bla(NDM-1) remains a challenge when using only phenotypic assays. Microbiology laboratories must be alert for K. pneumoniae isolates producing both KPC-2 and NDM-1.
  • article 0 Citação(ões) na Scopus
    Conjugative transfer of plasmid p_8N_qac(MN687830.1) carrying qacA gene from Staphylococcus aureus to Escherichia coli C600: potential mechanism for spreading chlorhexidine resistance
    (2021) BES, Taniela Marli; NAGANO, Debora Satie; MARCHI, Ana Paula; CAMILO, Gaspar; PERDIGAO-NETO, Lauro Vieira; MARTINS, Roberta Ruedas; LEVIN, Anna Sara; COSTA, Silvia Figueiredo
    The methicillin resistant Staphylococcus aureus (MRSA) is recognized by its ability to acquire and transferring resistance genes through interspecies conjugative plasmids. However, transference of plasmids from Gram-positive cocci to Gram-negative bacilli is not well characterized. In this report, we describe the transfer of a conjugative plasmid carrying qacA from MRSA to Escherichia coli C600. We performed a conjugation experiment using a chlorhexidine resistant MRSA isolate (ST-105/SCCmec type III) carrying the gene qacA and qacC as the donor and a chlorhexidine susceptible E. coli C600 isolate as the receptor. Transconjugants were selected using MacConkey agar plates containing chlorhexidine in concentrations ranging from 0.25 to 16 g.L-1. To genotypically confirm the transfer of the resistance gene, the transconjugants were screened by Polymerase Chain Reaction (PCR) and submitted to Sanger's sequencing. MRSA isolates successfully transferred the chlorhexidine resistance gene (qacA) to the recipient E. coli strain C600. The E. coli transconjugant exhibited an important reduction of chlorhexidine susceptibility, with MICs increasing from <= 0.25 to >= 16 g.L-1 after conjugation. The qacA gene was detected by PCR as well as in the Sanger's sequencing analysis of DNA from transconjugant plasmids. To the best of our knowledge, this is the first report of the plasmid p_8N_qac(MN687830.1) carrying qacA and its transfer by conjugation from a MRSA to an E. coli. These findings increase concerns on the emergence of resistance dissemination across the genus and emphasizes the importance of continuous antiseptic stewardship.