title: PCR-based identification and strain typing of Pichia anomala using the ribosomal intergenic spacer region IGS1.
abstract: Frequent outbreaks of Pichia anomala fungaemia in paediatric patients have warranted the development of a rapid identification system for this organism. This study describes a specific PCR-based method targeting the rRNA gene intergenic spacer region 1 (IGS1) for rapid identification of Pichia anomala isolates and characterization at the strain level. These methods of species identification and strain typing were used on 106 isolates of Pichia anomala (77 from a previously described outbreak and 29 isolated post-outbreak from the same hospital). Using conventional morphological and biochemical methods, 11 strains isolated during the outbreak were misidentified as P. anomala. blast analysis of sequences of internal transcribed spacer (ITS) regions of rRNA genes confirmed that they were Pichia guilliermondii (eight isolates) and Debaryomyces hansenii (three isolates). Strain typing of Pichia anomala isolates confirmed the previous finding of a point-source outbreak. The results suggest that IGS sequences and their polymorphisms could be exploited for similar typing methods in other organisms.
authors.affiliation: Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh 160012, India.
authors.forename: Sonia
authors.initials: S
authors.lastname: Bhardwaj
authors.affiliation: Institute of Microbial Technology, Sector 39-A, Chandigarh 160036, India.
authors.forename: Rajeshwari
authors.initials: R
authors.lastname: Sutar
authors.affiliation: Institute of Microbial Technology, Sector 39-A, Chandigarh 160036, India.
authors.forename: Anand K
authors.initials: AK
authors.lastname: Bachhawat
authors.affiliation: Department of Paediatrics, Postgraduate Institute of Medical Education and Research, Chandigarh 160012, India.
authors.forename: Sunit
authors.initials: S
authors.lastname: Singhi
authors.affiliation: Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh 160012, India.
authors.forename: Arunaloke
authors.initials: A
authors.lastname: Chakrabarti
mesh_terms: D004271:DNA, Fungal / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D004196:Disease Outbreaks; D004587:Electrophoresis, Agar Gel; D016469:Fungemia / Q000175:diagnosis / Q000382:microbiology; D020459:Genes, rRNA; D006801:Humans; D009181:Mycoses / Q000175:diagnosis / Q000382:microbiology*; D010843:Pichia / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer