title: New Delhi metallo-β-lactamase and extended spectrum β-lactamases co-producing isolates are high in community-acquired urinary infections in Assam as detected by a novel multiplex polymerase chain reaction assay.
abstract: BACKGROUND
  The ability of microorganisms to evade antibiotic pressure is challenging in healthcare as patients have little or no drug treatment options. Detection of the prevalence of antibacterial resistance pattern helps towards improved antibiotic policy and empirical treatment.
  
  
  OBJECTIVE
  We carried out antibiogram profiling and documented the prevalence and co-prevalence of New Delhi metallo-β-lactamase (NDM) and extended spectrum β-lactamases (ESBL) encoding genes in urinary Escherichia coli and Klebsiella pneumonia isolates.
  
  
  METHODS
  Antibiotic susceptibilities were tested for 241 isolates of E. coli and K. pneumoniae from urine samples collected from out- and hospitalised patients. Polymerase chain reaction (PCR) was carried out on isolates tested positive for phenotypic production of metallo-β-lactamase and ESBL. A multiplex PCR assay was designed to detect the genes.
  
  
  RESULTS
  Multiplex PCR assay designed had a limit of detection of 10 3 CFU/mL in vitro. NDM detected was significantly higher among K. pneumoniae compared to E. coli (69.2% vs. 18.2%; P = 0.001). Of 17, 14 NDM positive isolates also harboured ESBL genes. The co-production of CTX-M + TEM + NDM (3/9; 33.3% and 5/8; 62.5%) was most common in K. pneumoniae and E. coli, respectively while CTX-M + TEM + SHV + NDM was found in one isolate. Of the 156 phenotypically ESBL producing isolates, CTX-M, TEM and SHV was detected by PCR in 85, 53 and 24 isolates, respectively.
  
  
  CONCLUSIONS
  NDM and ESBL co-producing isolates were both community (64.7%) and hospital (35.29%) acquired among E. coli. Antibiotic resistance can be effectively evaluated by a cost and time effective molecular method, such as the multiplex PCR used in this study, which complement culture and sensitivity tests.
authors.forename: V V
authors.initials: VV
authors.lastname: Borah
authors.affiliation: Department of Bioengineering and Technology, Gauhati University, Guwahati, Assam, India.
authors.forename: K K
authors.initials: KK
authors.lastname: Saikia
authors.forename: P
authors.initials: P
authors.lastname: Chandra
authors.forename: N K
authors.initials: NK
authors.lastname: Hazarika
authors.forename: R
authors.initials: R
authors.lastname: Chakravarty
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D017714:Community-Acquired Infections / Q000453:epidemiology / Q000382:microbiology*; D004756:Enterobacteriaceae Infections / Q000453:epidemiology / Q000382:microbiology*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007194:India; D007223:Infant; D007711:Klebsiella pneumoniae / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D008297:Male; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction; D015995:Prevalence; D014552:Urinary Tract Infections / Q000453:epidemiology / Q000382:microbiology*; D055815:Young Adult; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D001618:beta-Lactamases