title: Phenotypic and molecular detection of the blaKPCgene in clinical isolates from inpatients at hospitals in São Luis, MA, Brazil.
abstract: Bacteria that produce Klebsiella pneumoniae carbapenemases (KPCs) are resistant to broad-spectrum β-lactam antibiotics. The objective of this study was to phenotypically and genotypically characterize the antibiotic susceptibility to carbapenems of 297 isolates recovered from clinical samples obtained from inpatients at 16 hospitals in São Luis (Maranhão, Brazil).
  
  
  
  The study was conducted using phenotypic tests and molecular methods, including polymerase chain reaction (PCR), sequencing and enterobacterial repetitive intergenic consensus (ERIC)-PCR. The nonparametric chi-square test of independence was used to evaluate the associations between the bacterial blaKPCgene and the modified Hodge test, and the chi-square adherence test was used to assess the frequency of carbapenemases and their association with the blaKPCgene.
  
  
  
  The most frequently isolated species were Acinetobacter baumannii (n = 128; 43.0%), K. pneumoniae (n = 75; 25.2%), and Pseudomonas aeruginosa (n = 42; 14.1%). Susceptibility assays showed that polymixin B was active against 89.3% of the bacterial isolates. The Acinetobacter spp. and K. pneumoniae strains were susceptible to amikacin and tigecycline, and Pseudomonas spp. were sensitive to gentamicin and amikacin. Among the 297 isolates, 100 (33.7%) were positive for the blaKPCgene, including non-fermentative bacteria (A. baumannii) and Enterobacteriaceae species. Among the isolates positive for the blaKPCgene, K. pneumoniae isolates had the highest positivity rate of 60.0%. The blaKPCgene variants detected included KPC-2, which was found in all isolates belonging to species of the Enterobacteriaceae family. KPC-2 and KPC-3 were observed in A. baumannii isolates. Importantly, the blaKPCgene was also detected in three Raoultella isolates and one isolate of the Pantoea genus. ERIC-PCR patterns showed a high level of genetic diversity among the bacterial isolates; it was capable of distinguishing 34 clones among 100 strains that were positive for blaKPCand were circulating in 11 of the surveyed hospitals.
  
  
  
  The high frequency of the blaKPCgene and the high degree of clonal diversity among microorganisms isolated from patients from different hospitals in São Luis suggest the need to improve the quality of health care to reduce the incidence of infections and the emergence of carbapenem resistance in these bacteria as well as other Gram-negative pathogens.
authors.affiliation: Programa de Pós-Graduação em Biologia Parasitária, Universidade CEUMA, Rua Josué Montello, No. 1, Renascença II, São Luís, Maranhão, CEP 65075-120, Brazil.
authors.forename: Patricia Cristina Saldanha
authors.initials: PC
authors.lastname: Ribeiro
authors.affiliation: Programa de Pós-Graduação em Biologia Parasitária, Universidade CEUMA, Rua Josué Montello, No. 1, Renascença II, São Luís, Maranhão, CEP 65075-120, Brazil.
authors.forename: Andrea Souza
authors.initials: AS
authors.lastname: Monteiro
authors.affiliation: Hospital Universitário da Universidade Federal do Maranhão, Rua Barão de Itapari, 227, Centro, São Luís, Maranhão, Brazil.
authors.forename: Sirlei Garcia
authors.initials: SG
authors.lastname: Marques
authors.affiliation: Programa de Pós-Graduação em Biologia Parasitária, Universidade CEUMA, Rua Josué Montello, No. 1, Renascença II, São Luís, Maranhão, CEP 65075-120, Brazil.
authors.forename: Sílvio Gomes
authors.initials: SG
authors.lastname: Monteiro
authors.affiliation: Programa de Pós-Graduação em Biologia Parasitária, Universidade CEUMA, Rua Josué Montello, No. 1, Renascença II, São Luís, Maranhão, CEP 65075-120, Brazil.
authors.forename: Valério
authors.initials: V
authors.lastname: Monteiro-Neto
authors.affiliation: Programa de Pós-Graduação em Biologia Parasitária, Universidade CEUMA, Rua Josué Montello, No. 1, Renascença II, São Luís, Maranhão, CEP 65075-120, Brazil.
authors.forename: Martina Márcia Melo
authors.initials: MM
authors.lastname: Coqueiro
authors.affiliation: Programa de Pós-Graduação em Saúde do Adulto e da Criança-UFMA, Universidade Federal do Maranhão, Av. dos Portugueses, 1966 - Bacanga, São Luis, Maranhão, Brazil.
authors.forename: Ana Cláudia Garcia
authors.initials: AC
authors.lastname: Marques
authors.affiliation: Departamento de Farmácia, Universidade Federal do Maranhão, Av. dos Portugueses, 1966 - Bacanga, São Luis, Maranhão, Brazil.
authors.forename: Rosimary
authors.initials: R
authors.lastname: de Jesus Gomes Turri
authors.affiliation: Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos, 6627, Pampulha, Belo Horizonte, MG, Brazil.
authors.forename: Simone Gonçalves
authors.initials: SG
authors.lastname: Santos
authors.affiliation: Programa de Pós-Graduação em Biologia Parasitária, Universidade CEUMA, Rua Josué Montello, No. 1, Renascença II, São Luís, Maranhão, CEP 65075-120, Brazil. mrqbomfim@yahoo.com.br.
authors.forename: Maria Rosa Quaresma
authors.initials: MR
authors.lastname: Bomfim
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001938:Brazil; D015780:Carbapenems / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000187:drug effects / Q000235:genetics; D004755:Enterobacteriaceae / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D006761:Hospitals; D006801:Humans; D007297:Inpatients; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D008826:Microbial Sensitivity Tests; D017720:Molecular Epidemiology / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism; D047090:beta-Lactams / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D015780:Carbapenems; D047090:beta-Lactams; D001618:beta-Lactamases; C063912:carbapenemase
keywords: Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR); Klebsiella pneumoniae carbapenemases (KPCs); Multidrug-resistance (MDR); bla KPC gene variants