title: Development of two molecular approaches for differentiation of clinically relevant yeast species closely related to Candida guilliermondii and Candida famata.
abstract: The emerging pathogens Candida palmioleophila, Candida fermentati, and Debaryomyces nepalensis are often misidentified as Candida guilliermondii or Candida famata in the clinical laboratory. Due to the significant differences in antifungal susceptibilities and epidemiologies among these closely related species, a lot of studies have focused on the identification of these emerging yeast species in clinical specimens. Nevertheless, limited tools are currently available for their discrimination. Here, two new molecular approaches were established to distinguish these closely related species. The first approach differentiates these species by use of restriction fragment length polymorphism analysis of partial internal transcribed spacer 2 (ITS2) and large subunit ribosomal DNA with the enzymes BsaHI and XbaI in a double digestion. The second method involves a multiplex PCR based on the intron size differences of RPL18, a gene coding for a protein component of the large (60S) ribosomal subunit, and species-specific amplification. These two methods worked well in differentiation of these closely related yeast species and have the potential to serve as effective molecular tools suitable for laboratory diagnoses and epidemiological studies.
authors.affiliation: Medical Mycology Laboratory, Department of Dermatology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Feng
authors.affiliation: Department of Laboratory Medicine, Shenzhen People's Hospital, Second Clinical Medical College of Jinan University, Shenzhen, People's Republic of China.
authors.forename: Jingsong
authors.initials: J
authors.lastname: Wu
authors.affiliation: Medical Mycology Laboratory, Department of Dermatology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Ling
authors.affiliation: Medical Mycology Laboratory, Department of Dermatology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China.
authors.forename: Xianwei
authors.initials: X
authors.lastname: Yang
authors.affiliation: Shanghai Key Laboratory of Molecular Medical Mycology, Shanghai Institute of Medical Mycology, Changzheng Hospital, Second Military Medical University, Shanghai, People's Republic of China.
authors.forename: Wanqing
authors.initials: W
authors.lastname: Liao
authors.affiliation: Shanghai Key Laboratory of Molecular Medical Mycology, Shanghai Institute of Medical Mycology, Changzheng Hospital, Second Military Medical University, Shanghai, People's Republic of China panweihua@medmail.com.cn dermatology.yao@sohu.com.
authors.forename: Weihua
authors.initials: W
authors.lastname: Pan
authors.affiliation: Medical Mycology Laboratory, Department of Dermatology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People's Republic of China panweihua@medmail.com.cn dermatology.yao@sohu.com.
authors.forename: Zhirong
authors.initials: Z
authors.lastname: Yao
mesh_terms: D004271:DNA, Fungal / Q000235:genetics*; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology*; D012150:Polymorphism, Restriction Fragment Length; D012335:RNA, Ribosomal / Q000235:genetics; D004718:Saccharomycetales / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004271:DNA, Fungal; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D005656:Fungal Proteins; D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S