, Volume 178, Issue 1–2, pp 53–61 | Cite as

The Application of Laser Microdissection in Molecular Detection and Identification of Aspergillus fumigatus from Murine Model of Acute Invasive Pulmonary Aspergillosis

  • Chong Wang
  • Ping Zhan
  • Le Wang
  • Rong Zeng
  • Yongnian Shen
  • Guixia Lv
  • Dongmei Li
  • Shuwen Deng
  • Weida Liu


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.


Invasive aspergillosis Laser capture microdissection PCR Murine model 



This investigation was supported by Youth Innovative Project (2012x13) from Peking Union Medical College, by Major National Science and Technology Projects (2013ZX10004612-005) from Ministry of Science and Technology of P.R.C., by Jiangsu Provincial Special Program of Medical Science (BL2012003) from Science and Technology Department of Jiangsu Province, and by key project of the ministry of health, clinical disciplines (2010–2012) from the ministry of health of P.R.C.

Conflict of interest

The authors declare that they have no conflict of interests.


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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Chong Wang
    • 1
    • 2
  • Ping Zhan
    • 1
    • 2
  • Le Wang
    • 1
    • 2
  • Rong Zeng
    • 1
    • 2
  • Yongnian Shen
    • 1
    • 2
  • Guixia Lv
    • 1
    • 2
  • Dongmei Li
    • 3
  • Shuwen Deng
    • 4
  • Weida Liu
    • 2
    • 1
  1. 1.Department of Mycology, Institute of DermatologyChinese Academy of Medical Science and Peking Union Medical CollegeNanjingPeople’s Republic of China
  2. 2.Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIsNanjingPeople’s Republic of China
  3. 3.Department of Microbiology and ImmunologyGeorgetown University Medical CenterWashingtonUSA
  4. 4.Shanghai Institute of Medical Mycology, Changzheng HospitalSecond Military Medical UniversityShanghaiPeople’s Republic of China

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