KMID : 0545120140240081044
|
|
Journal of Microbiology and Biotechnology 2014 Volume.24 No. 8 p.1044 ~ p.1050
|
|
Evaluation of Galactomannan Enzyme Immunoassay and Quantitative Real-Time PCR for the Diagnosis of Invasive Pulmonary Aspergillosis in a Rat Model
|
|
Jian-cong Lin
Yan-li Xing Wen-ming Xu Ming Li Pang Bo Yuan-yuan Niu Chang-ran Zhang
|
|
Abstract
|
|
|
Since there is no consensus about the most reliable assays to detect invasive aspergillosis from samples obtained by minimally invasive or noninvasive methods, we compared the efficacy of an enzyme-linked immunosorbent assay (ELISA) for galactomannan (GM) detection and quantitative real-time PCR assay (qRT-PCR) for the diagnosis of invasive pulmonary aspergillosis. Neutropenic, male Sprague-Dawley rats (specific pathogen free; 8 weeks old; weight, 200 ¡¾ 20 g) were immunosuppressed with cyclophosphamide and infected with Aspergillus fumigatus intratracheally. Tissue and whole blood samples were harvested on days 1, 3, 5, and 7 post-infection and examined with GM ELISA and qRT-PCR. The A. fumigatus DNA detection sequence was detected in the following number of samples from 12 immunosuppressed, infected rats examined on the scheduled days: day 1 (0/12), day 3 (0/12), day 5 (6/12), and day 7 (8/12) post-infection. The sensitivity and specificity of the qRT-PCR assay was 29.2% and 100%, respectively. Receiver operating characteristic curve (ROC) analysis indicated a Ct (cycle threshold) cut-off value of 15.35, and the area under the curve (AUC) was 0.627. The GM assay detected antigen in sera obtained on day 1 (5/12), day 3 (9/ 12), day 5 (12/12), and day 7 (12/12) post-infection, and thus had a sensitivity of 79.2% and a specificity of 100%. The ROC of the GM assay indicated that the optimal Ct cut-off value was 1.40 (AUC, 0.919). The GM assay was more sensitive than the qRT-PCR assay in diagnosing invasive pulmonary aspergillosis in rats.
|
|
KEYWORD
|
|
Invasive pulmonary aspergillosis, Aspergillus, galactomannan antigen, Quantitative real-time PCR, Receiver operating characteristic curve, rat model
|
|
FullTexts / Linksout information
|
|
|
|
Listed journal information
|
|
|
|