title: Three isothermal amplification techniques for rapid identification of Cladophialophora carrionii, an agent of human chromoblastomycosis.
abstract: In this study, we developed rapid and sensitive assays for the detection of Cladophialophora carrionii, a common agent of human chromoblastomycosis. The isothermal techniques evaluated were rolling-circle amplification (RCA), multiplex ligation-dependent probe amplification (MLPA), and loop-mediated isothermal amplification (LAMP). The probes for RCA and MLPA were designed with target sequences in the rDNA internal transcribed spacer gene (ITS) region, and LAMP primers were designed using the elongation factor 1α gene (EF1); these probes and primers specifically amplified DNA of isolates of the species. The three techniques were sufficiently specific and sensitive for discriminating target DNA of C. carrionii from that of related Cladophialophora species and other agents of chromoblastomycosis. RCA, MLPA, and LAMP are advantageous in their reliability and ease of operation compared to standard PCR and conventional methods.
authors.affiliation: Shanghai Key Laboratory of Molecular Medical Mycology, Changzheng Hospital, Second Military Medical University, Shanghai, China Department of Dermatology, First Hospital of Xinjiang Medical University, Urumqi, Xinjiang, China CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands.
authors.forename: Shuwen
authors.initials: S
authors.lastname: Deng
authors.affiliation: CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands Shanghai Key Laboratory of Molecular Medical Mycology, Changzheng Hospital, Second Military Medical University, Shanghai, China Institute of Biodiversity and Ecosystem Dynamics, University of Amsterdam, Amsterdam, Netherlands Basic Pathology Department, Federal University of Paraná State, Curitiba, Paraná, Brazil King Abdulaziz University, Jeddah, Saudi Arabia Department of Dermatology, First Hospital of Peking University, Beijing, China.
authors.forename: G Sybren
authors.initials: GS
authors.lastname: de Hoog
authors.affiliation: Shanghai Key Laboratory of Molecular Medical Mycology, Changzheng Hospital, Second Military Medical University, Shanghai, China.
authors.forename: Weihua
authors.initials: W
authors.lastname: Pan
authors.affiliation: Shanghai Key Laboratory of Molecular Medical Mycology, Changzheng Hospital, Second Military Medical University, Shanghai, China.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
authors.affiliation: CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands.
authors.forename: A H G Gerrits
authors.initials: AH
authors.lastname: van den Ende
authors.affiliation: CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands.
authors.forename: Liyue
authors.initials: L
authors.lastname: Yang
authors.affiliation: Guangdong Provincial Institute of Public Health, Guangdong Provincial Centers for Disease Control and Prevention, Guangzhou, China.
authors.forename: Jiufeng
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Parasitology and Mycology, Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran CBS-KNAW Fungal Biodiversity Centre, Utrecht, Netherlands.
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Najafzadeh
authors.affiliation: Shanghai Key Laboratory of Molecular Medical Mycology, Changzheng Hospital, Second Military Medical University, Shanghai, China liaowanqing@sohu.com lry0660@gmail.com.
authors.forename: Wanqing
authors.initials: W
authors.lastname: Liao
authors.affiliation: Department of Dermatology, First Hospital of Peking University, Beijing, China liaowanqing@sohu.com lry0660@gmail.com.
authors.forename: Ruoyu
authors.initials: R
authors.lastname: Li
mesh_terms: D001203:Ascomycota / Q000235:genetics / Q000302:isolation & purification*; D002862:Chromoblastomycosis / Q000175:diagnosis* / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D020648:Peptide Elongation Factor 1; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013997:Time Factors
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D015345:Oligonucleotide Probes; D020648:Peptide Elongation Factor 1