ORIGINAL ARTICLE   

Minerva Pneumologica 2020 December;59(4):65-9

DOI: 10.23736/S0026-4954.20.01885-4

Copyright © 2020 EDIZIONI MINERVA MEDICA

language: English

Real-time PCR-based method for detection of Aspergillus Spp in bronchoalveolar respiratory samples

Somayeh SHARIFYNIA ¹, Payam TABARSI ¹, Naghmeh BAHRAMI ², Hamidreza JAMAATI ³, Mihan POURABDOLLAH ³, Elham ASKARI ³, Mahya DAUSTANY ⁴, Maral EMAMI ³, Sona ZARE ⁵, Yasaman HASANI ⁶, Abdolreza MOHAMADNIA ^{3, 7} ✉

¹ Clinical Tuberculosis and Epidemiology Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran; ² Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran; ³ Chronic Respiratory Diseases Research Center, National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran; ⁴ Department of Biotechnology, Faculty of Sciences, Islamic Azad University, Tehran, Iran; ⁵ Skin and Stem Cell Research Center, Tehran University of Medical Sciences, Tehran, Iran; ⁶ Private Practitioner, Tehran, Iran; ⁷ Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

BACKGROUND: Some of the Aspergillus species are known to cause infectious diseases in humans; also, invasive aspergillosis is life-threatening for individuals with immunodeficiency. Thus, precise and rapid detection of Aspergillus and identification of its species are crucial. To detect Aspergillus and identify Aspergillus species in clinical samples using real-time polymerase chain reaction (PCR).
METHODS: In this study, a total of 100 BAL samples were collected from patients who referred to Masih Daneshvari Hospital and sent to laboratory to be cultured. PCR and real-time PCR were performed for samples using specific primers. Real-time PCR served as a gold standard. For final approval, some of the samples sent for sequencing.
RESULTS: Ninety-five of the samples reported positive for fungi in the culture, 16 of these were positive for Aspergillus. In PCR, 94 samples were found positive for fungi, and 12 samples found to be Aspergillus. Finally, real-time PCR reported 97 positive cases for fungi and 18 for Aspergillus. Of these 18 cases, 6 Aspergillus flavus, 4 Aspergillus fumigatus and 3 Aspergillus terreus were isolated.
CONCLUSIONS: Real-time PCR was shown to be more accurate and rapid technique in comparison with culture test and conventional PCR in identification of Aspergillus and its species with high accuracy.

KEY WORDS: Aspergillus; Immunocompromised host; Real-time polymerase chain reaction