title: Molecular identification ofCandidaspecies isolated from cases of neonatal candidemia using polymerase chain reaction-restriction fragment length polymorphism in a tertiary care hospital.
abstract: BACKGROUND
  Candida spp. is an emerging cause of bloodstream infections worldwide. Delay in speciation of Candida isolates by conventional methods and resistance to antifungal drugs in various Candida species are responsible for the increase in morbidity and mortality due to candidemia. Hence, the rapid identification of Candida isolates is very important for the proper management of patients with candidemia.
  
  
  OBJECTIVE
  The aim was to re-evaluate the identification of various Candida spp. by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) and to evaluate the accuracy, speed, and cost of phenotypic methodology versus PCR-RFLP.
  
  
  METHODS
  Hospital-based cross-sectional study.
  
  
  METHODS
  Ninety consecutive clinical isolates of seven Candida species, isolated from blood of neonates and identified by routine phenotypic methods, were re-evaluated using universal primers internal transcribed spacer 1 (ITS1) and ITS4 for PCR amplification and Msp I restriction enzyme for RFLP.
  
  
  METHODS
  Kappa test for agreement.
  
  
  RESULTS
  The results of PCR-RFLP were 100% in agreement with those obtained using conventional phenotypic methods. Identification could be achieved within 3 work days by both the methods. Our routine methods proved to be cost effective than PCR-RFLP.
  
  
  CONCLUSIONS
  We can continue with our routine phenotypic methods and PCR-RFLP can be used for periodic quality control or when conventional methods fail to identify a species.
authors.affiliation: Department of Microbiology, Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Jammu and Kashmir, India.
authors.forename: Akeela
authors.initials: A
authors.lastname: Fatima
authors.affiliation: Department of Microbiology, Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Jammu and Kashmir, India.
authors.forename: Gulnaz
authors.initials: G
authors.lastname: Bashir
authors.affiliation: Department of Microbiology, Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Jammu and Kashmir, India.
authors.forename: Tehmeena
authors.initials: T
authors.lastname: Wani
authors.affiliation: Department of Microbiology, Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Jammu and Kashmir, India.
authors.forename: Abiroo
authors.initials: A
authors.lastname: Jan
authors.affiliation: Department of Microbiology, Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Jammu and Kashmir, India.
authors.forename: Amrish
authors.initials: A
authors.lastname: Kohli
authors.affiliation: Department of Clinical Biochemistry, Sher-I-Kashmir Institute of Medical Sciences, Srinagar, Jammu and Kashmir, India.
authors.forename: Mosin S
authors.initials: MS
authors.lastname: Khan
mesh_terms: D002175:Candida / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D058387:Candidemia / Q000175:diagnosis* / Q000382:microbiology*; D003365:Costs and Cost Analysis; D003430:Cross-Sectional Studies; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D006801:Humans; D007231:Infant, Newborn; D008828:Microbiological Techniques / Q000191:economics / Q000379:methods; D025202:Molecular Diagnostic Techniques / Q000191:economics / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D062606:Tertiary Care Centers; D013997:Time Factors
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer