半乳甘露聚糖酶联免疫吸附测定（ELISA）和聚合酶链反应（PCR）在变应性支气管肺曲霉病（ABPA）和慢性肺曲霉病（CPA）患者痰液中的比较表现

Comparative performance of Aspergillus galactomannan ELISA and PCR in sputum from patients with ABPA and CPA.

作者信息

Fayemiwo Samuel, Moore Caroline B, Foden Philip, Denning David W, Richardson Malcolm D

机构信息

Department of Medical Microbiology and Parasitology, College of Medicine, University of Ibadan, Ibadan, Nigeria; Mycology Reference Centre, University Hospital of South Manchester, Manchester Academic Health Sciences Centre, Manchester, UK; Institute of inflammation and Repair, Faculty of Medicine and Health Sciences, University of Manchester, UK.

Mycology Reference Centre, University Hospital of South Manchester, Manchester Academic Health Sciences Centre, Manchester, UK; Institute of inflammation and Repair, Faculty of Medicine and Health Sciences, University of Manchester, UK.

出版信息

J Microbiol Methods. 2017 Sep;140:32-39. doi: 10.1016/j.mimet.2017.06.016. Epub 2017 Jun 21.

DOI:10.1016/j.mimet.2017.06.016

PMID:28645480

Abstract

OBJECTIVES

Galactomannan (GM) and Aspergillus DNA detection are useful tools for the diagnosis of invasive pulmonary aspergillosis (IPA), primarily in blood and bronchoscopy samples. This study aimed to evaluate the utility of both markers for detection of Aspergillus in sputum from patients with allergic bronchopulmonary aspergillosis (ABPA) and chronic pulmonary aspergillosis (CPA).

METHODS

ABPA or CPA demographic patient data were retrieved. This retrospective observational audit included 159 patients with at least one sputum pair. 223 sputum sample pairs were analysed, as well as six control samples for GM only. Real time PCR was performed following sputum DNA extraction using the MycAssay™ Aspergillus kit and cycle thresholds were subtracted from 38 to give positive values (transformed Ct, TCt).

RESULTS

The mean age of the patients was 61.81years (SD: ±11.06; range 29-100). One hundred and twenty-six (79.2%) had CPA. Cultures were positive for fungi in 13.1% of the samples, and A. fumigatus was the commonest (11.9%) fungus isolated. Receiver operating characteristic (ROC curve) analysis of sputum GM comparing TCt of >0.0, and >2.0 to derive GMI cut-off values showed a cut-off of 6.5. About 50% of sputa with strongly positive PCR values had GM values>6.5. Two of six (33%) control samples had GM indices>6.5.

CONCLUSION

It is not clear that GM determinations in sputum are useful for diagnosis of either CPA or ABPA, or following therapy.

摘要

目的

半乳甘露聚糖（GM）和曲霉DNA检测是诊断侵袭性肺曲霉病（IPA）的有用工具，主要用于血液和支气管镜检查样本。本研究旨在评估这两种标志物在过敏性支气管肺曲霉病（ABPA）和慢性肺曲霉病（CPA）患者痰液中检测曲霉的效用。

方法

检索ABPA或CPA患者的人口统计学数据。这项回顾性观察性审计纳入了159例至少有一对痰液样本的患者。分析了223对痰液样本以及仅用于GM检测的6个对照样本。使用MycAssay™曲霉试剂盒提取痰液DNA后进行实时PCR，并将循环阈值从38中减去以得到正值（转化Ct，TCt）。

结果

患者的平均年龄为61.81岁（标准差：±11.06；范围29 - 100岁）。126例（79.2%）患有CPA。13.1%的样本培养出真菌阳性，烟曲霉是最常见的分离真菌（11.9%）。对痰液GM进行受试者操作特征（ROC曲线）分析，比较TCt>0.0和>2.0以得出GM指数（GMI）临界值，结果显示临界值为6.5。PCR值强阳性的痰液中约50%的GM值>6.5。6个对照样本中有2个（33%）的GM指数>6.5。