AUTHOR=Jiang Xiaolin , Jiang Yong , Ye Feng 

TITLE=Detection and identification of Mucorales and Aspergillus in paraffin-embedded samples by real-time quantitative PCR

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=13

YEAR=2023

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2023.1082347

DOI=10.3389/fcimb.2023.1082347

ISSN=2235-2988

ABSTRACT=<sec><title>Background</title><p>In this study, we used real-time quantitative PCR (RQ-PCR) to rapidly detect <italic>Mucorales</italic> and <italic>Aspergillus</italic> in formalin-fixed, paraffin-embedded (FFPE) samples, targeting 18SrRNA gene and 28SrRNA gene. Identification of <italic>Mucorales</italic> and <italic>Aspergillus</italic> was analysed by combining <italic>Mucorales</italic> RQ-PCR (<italic>Mucorales</italic>18SrRNA and <italic>Mucorales</italic>28SrRNA) with <italic>Aspergillus</italic> RQ-PCR (<italic>Aspergillus</italic>18SrRNA and <italic>Aspergillus</italic>28SrRNA).</p></sec><sec><title>Objectives</title><p>The aims of this study were to compare the diagnostic performances of four RQ-PCR assays as single and combined diagnostic and identification tools.</p></sec><sec><title>Methods</title><p>We collected 12 control group samples and 81 experimental group samples diagnosed by histopathology, including mucormycosis (19 patients, 21 FFPE samples), aspergillosis (54 patients, 57 FFPE samples) and mucormycosis with aspergillosis (3 patients, 3 FFPE samples). All samples were detected by four RQ-PCR tests to compare and analyze diagnostic performance.</p></sec><sec><title>Results</title><p>The sensitivities of <italic>Mucorales</italic>18SrRNA and <italic>Mucorales</italic>28SrRNA were both 75%, with the tests having specificities of 97.10% and 94.20%. The sensitivities of <italic>Aspergillus</italic>18SrRNA and <italic>Aspergillus</italic>28SrRNA were 73.33% and 65%, with the tests having specificities of 87.88% and 81.82%. The values of the evaluation indexes of the combined detection of <italic>Mucorales</italic>28SrRNA and <italic>Aspergillus</italic>18SrRNA (M28A18) were the highest with a kappa coefficient value of 0.353, followed by M18A18. M28A18 had a sensitivity of 67.90% and a specificity of 100%.</p></sec><sec><title>Conclusions</title><p>We recommend using the combination of <italic>Mucorales</italic> RQ-PCR and <italic>Aspergillus</italic> RQ-PCR as a screening tool to detect samples suspected of mucormycosis and/or aspergillosis.</p></sec>