应用实时定量 PCR 技术对石蜡包埋样本中 和 的检测与鉴定。
Detection and identification of and in paraffin-embedded samples by real-time quantitative PCR.
机构信息
Department of Pathology, West China Hospital of Sichuan University, Chengdu, China.
Department of Pathology, Guangyuan Central Hospital, Guangyuan, China.
出版信息
Front Cell Infect Microbiol. 2023 Mar 2;13:1082347. doi: 10.3389/fcimb.2023.1082347. eCollection 2023.
BACKGROUND
In this study, we used real-time quantitative PCR (RQ-PCR) to rapidly detect and in formalin-fixed, paraffin-embedded (FFPE) samples, targeting 18SrRNA gene and 28SrRNA gene. Identification of and was analysed by combining RQ-PCR (18SrRNA and 28SrRNA) with RQ-PCR (18SrRNA and 28SrRNA).
OBJECTIVES
The aims of this study were to compare the diagnostic performances of four RQ-PCR assays as single and combined diagnostic and identification tools.
METHODS
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.
RESULTS
The sensitivities of 18SrRNA and 28SrRNA were both 75%, with the tests having specificities of 97.10% and 94.20%. The sensitivities of 18SrRNA and 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 28SrRNA and 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%.
CONCLUSIONS
We recommend using the combination of RQ-PCR and RQ-PCR as a screening tool to detect samples suspected of mucormycosis and/or aspergillosis.
背景
本研究采用实时定量 PCR（RQ-PCR）技术，以 18SrRNA 基因和 28SrRNA 基因为靶标，快速检测福尔马林固定、石蜡包埋（FFPE）样本中的 和 。通过将 18SrRNA 和 28SrRNA 的 RQ-PCR 与 18SrRNA 和 28SrRNA 的 RQ-PCR 相结合，分析 和 的鉴定结果。
目的
本研究旨在比较四种 RQ-PCR 检测方法作为单一和联合诊断及鉴定工具的诊断性能。
方法
我们收集了 12 例对照组样本和 81 例经组织病理学诊断的实验组样本，包括毛霉病（19 例患者，21 例 FFPE 样本）、曲霉病（54 例患者，57 例 FFPE 样本）和毛霉病合并曲霉病（3 例患者，3 例 FFPE 样本）。对所有样本进行了四种 RQ-PCR 检测，以比较和分析诊断性能。
结果
18SrRNA 和 28SrRNA 的灵敏度均为 75%，特异性分别为 97.10%和 94.20%。18SrRNA 和 28SrRNA 的灵敏度分别为 73.33%和 65%，特异性分别为 87.88%和 81.82%。28SrRNA 和 18SrRNA 联合检测（M28A18）的评价指标值最高，kappa 系数值为 0.353，其次是 M18A18。M28A18 的灵敏度为 67.90%，特异性为 100%。
结论
我们建议使用 18SrRNA 和 28SrRNA 的 RQ-PCR 联合检测作为一种筛选工具，用于检测疑似毛霉病和/或曲霉病的样本。
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