title: The application of laser microdissection in molecular detection and identification of aspergillus fumigatus from murine model of acute invasive pulmonary aspergillosis.
abstract: Invasive aspergillosis (IA) is a major concern in patients with severe immune deficiency. As antifungal susceptibility varies in different fungal pathogens, accurate and timely identification of species is becoming imperative for guidance of therapy and reducing high mortality rates in patients with IA. But, in fact, the diagnosis is challenging and new validated techniques are required for the detection and identification of clinically relevant isolates. The laser capture microdissection (LCM) system enables analysis of cytologically and/or phenotypically defined cell types from heterogeneous tissue and has been used in diagnosis and fungal species identification in pulmonary aspergillosis of white storks. To establish the experimental foundation for clinical application of the system, we microdissected and collected Blankophor-stained single hyphal strands from tissue cryosections of murine model of invasive pulmonary aspergillosis (IPA) with A. fumigatus by LCM, subsequently processed for DNA extraction, PCR sequencing, and species molecular identification. The sensitivity of LCM-PCR sequencing was 89 % (89/100), and the specificity was 100 %. Moreover, the positive predictive value and negative predictive value were 100 and 78.43 %, respectively. The result approved that the LCM-based methods had the potential for accurately diagnosis and rapidly identification fungal pathogens of IPA.
authors: Department of Mycology, Institute of Dermatology, Chinese Academy of Medical Science and Peking Union Medical College, No. 12, Jiang Wangmiao Street, Nanjing, 210042, People's Republic of China.
authors: Chong
authors: C
authors: Wang
authors: Ping
authors: P
authors: Zhan
authors: Le
authors: L
authors: Wang
authors: Rong
authors: R
authors: Zeng
authors: Yongnian
authors: Y
authors: Shen
authors: Guixia
authors: G
authors: Lv
authors: Dongmei
authors: D
authors: Li
authors: Shuwen
authors: S
authors: Deng
authors: Weida
authors: W
authors: Liu
mesh_terms: D000818:Animals; D001232:Aspergillus fumigatus / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004195:Disease Models, Animal; D055744:Invasive Pulmonary Aspergillosis / Q000175:diagnosis*; D061346:Laser Capture Microdissection / Q000379:methods*; D008807:Mice, Inbred BALB C; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D004271:DNA, Fungal