通过特异性和广谱实时聚合酶链反应从组织块中检测荚膜组织胞浆菌DNA：阐明组织胞浆菌病流行病学的工具

Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis.

作者信息

Wilmes Dunja, McCormick-Smith Ilka, Lempp Charlotte, Mayer Ursula, Schulze Arik Bernard, Theegarten Dirk, Hartmann Sylvia, Rickerts Volker

机构信息

Reference Laboratory for Cryptococcosis and Uncommon Invasive Fungal Infections, Division for Mycotic and Parasitic Agents and Mycobacteria, Robert Koch Institute, 13353 Berlin, Germany.

Vet Med Labor GmbH, Division of IDEXX Laboratories, 71636 Ludwigsburg, Germany.

出版信息

J Fungi (Basel). 2020 Nov 27;6(4):319. doi: 10.3390/jof6040319.

DOI:10.3390/jof6040319

PMID:33261008

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7711923/

Abstract

Lack of sensitive diagnostic tests impairs the understanding of the epidemiology of histoplasmosis, a disease whose burden is estimated to be largely underrated. Broad-range PCRs have been applied to identify fungal agents from pathology blocks, but sensitivity is variable. In this study, we compared the results of a specific qPCR (. qPCR) with the results of a broad-range qPCR (28S qPCR) on formalin-fixed, paraffin-embedded (FFPE) tissue specimens from patients with proven fungal infections ( = 67), histologically suggestive of histoplasmosis ( = 36) and other mycoses ( = 31). The clinical sensitivity for histoplasmosis of the . qPCR and the 28S qPCR was 94% and 48.5%, respectively. Samples suggestive for other fungal infections were negative with the . qPCR. The 28S qPCR did not amplify DNA of in FFPE in these samples, but could amplify DNA of ( = 1) and ( = 2) in three samples suggestive for histoplasmosis but negative in the qPCR. In conclusion, amplification of DNA from FFPE samples is more sensitive with the qPCR than with the 28S qPCR. However, the 28S qPCR identified DNA of other fungi in . qPCR-negative samples presenting like histoplasmosis, suggesting that the combination of both assays may improve the diagnosis.

摘要

缺乏敏感的诊断测试妨碍了对组织胞浆菌病流行病学的了解，据估计，该病的负担被大大低估。广谱聚合酶链反应（PCR）已被用于从病理切片中鉴定真菌病原体，但灵敏度各不相同。在本研究中，我们比较了特异性定量聚合酶链反应（. qPCR）与广谱定量聚合酶链反应（28S qPCR）对确诊真菌感染患者（n = 67）、组织学提示组织胞浆菌病患者（n = 36）和其他霉菌病患者（n = 31）的福尔马林固定石蜡包埋（FFPE）组织标本的检测结果。. qPCR和28S qPCR对组织胞浆菌病的临床灵敏度分别为94%和48.5%。提示其他真菌感染的样本在. qPCR检测中呈阴性。28S qPCR在这些样本的FFPE中未扩增出[具体真菌名称]的DNA，但在三个提示组织胞浆菌病但. qPCR检测为阴性的样本中扩增出了[具体真菌名称1]（n = 1）和[具体真菌名称2]（n = 2）的DNA。总之，从FFPE样本中扩增[具体真菌名称]DNA，. qPCR比28S qPCR更敏感。然而，28S qPCR在表现类似组织胞浆菌病的. qPCR阴性样本中鉴定出了其他真菌的DNA，这表明两种检测方法结合可能会改善诊断。