一种用于诊断烟曲霉感染的聚合酶链反应酶免疫测定法。
A polymerase chain reaction enzyme immunoassay for diagnosing infection caused by Aspergillus fumigatus.
作者信息
Golbang N, Burnie J P, Matthews R C
机构信息
Department of Medical Microbiology, Manchester University, Manchester Royal Infirmary, UK.
出版信息
J Clin Pathol. 1999 Jun;52(6):419-23. doi: 10.1136/jcp.52.6.419.
AIM
To develop a polymerase chain reaction enzyme immunoassay (PCR-EIA) to measure levels of circulating aspergillus DNA in invasive aspergillosis caused by Aspergillus fumigatus.
METHODS
The PCR reaction was based on primers from the 18s rRNA gene. Binding of the product to a streptavidin coated microtitration plate was mediated by a biotinylated capture probe. The product was digoxigenylated during PCR and this was the tag to which antibody was bound in the subsequent EIA.
RESULTS
The optical density (OD) endpoint was < 0.1 in 10 sera from neutropenic patients with no evidence of invasive aspergillosis, and in 10 sera from nonneutropenic patients with bacterial pneumonia (group 1). The OD from five of 12 patients with allergic bronchopulmonary aspergillosis (ABPA) (group 2), three with an aspergilloma (group 3), and five with possible invasive aspergillosis (group 4) was > or = 0.1. In 63 sera from 33 cases of proven invasive aspergillosis (group 5) an OD > or = 0.1 was achieved in 48 sera from 30 patients. The maximum OD was 0.510. The level fell in survivors and gradually rose in fatal cases.
CONCLUSIONS
This assay validated the concept of diagnosing invasive aspergillosis by measuring levels of circulating fungal DNA in serum.
目的
开发一种聚合酶链反应酶免疫测定法（PCR-EIA），以测量由烟曲霉引起的侵袭性曲霉病中循环曲霉DNA的水平。
方法
PCR反应基于18s rRNA基因的引物。产物与链霉亲和素包被的微量滴定板的结合由生物素化的捕获探针介导。产物在PCR过程中被地高辛标记，这是后续EIA中抗体结合的标签。
结果
10例无侵袭性曲霉病证据的中性粒细胞减少患者的血清以及10例细菌性肺炎的非中性粒细胞减少患者的血清（第1组）的光密度（OD）终点<0.1。12例变应性支气管肺曲霉病（ABPA）患者（第2组）中的5例、3例曲菌球患者（第3组）以及5例可能患有侵袭性曲霉病的患者（第4组）的OD≥0.1。在33例确诊的侵袭性曲霉病患者的63份血清（第5组）中，30例患者的48份血清的OD≥0.1。最大OD为0.510。幸存者的水平下降，而致命病例的水平逐渐上升。
结论
该测定法验证了通过测量血清中循环真菌DNA水平来诊断侵袭性曲霉病的概念。
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