Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Rapid Detection of Aspergillus fumigatus

作者：Tang, Qing; Tian, Shuguang; Yu, Nong; Zhang, Xi; Jia, Xiaodong; Zhai, Hongyan; Sun, Qun^*; Han, Li^*

来源：Journal of Clinical Microbiology, 2016, 54(4): 950-955.

DOI：10.1128/JCM.01751-15

• BRONCHOALVEOLAR LAVAGE FLUID
• INVASIVE PULMONARY ASPERGILLOSIS
• REAL-TIME PCR
• GALACTOMANNAN ENZYME-IMMUNOASSAY
• QUANTITATIVE PCR
• IMMUNOCOMPROMISED PATIENTS
• FUNGAL-INFECTIONS
• ASSAY
• DIAGNOSIS
• CONSENSUS

Aspergillus fumigatus is a conditional pathogen and the major cause of life-threatening invasive aspergillosis (IA) in immunocompromised patients. The early and rapid detection of A. fumigatus infection is still a major challenge. In this study, the new member of the fungal annexin family, annexin C4, was chosen as the target to design a loop-mediated isothermal amplification (LAMP) assay for the rapid, specific, and sensitive detection of A. fumigatus. The evaluation of the specificity of the LAMP assay that was developed showed that no false-positive results were observed for the 22 non-A. fumigatus strains, including 5 species of the Aspergillus genus. Its detection limit was approximately 10 copies per reaction in reference plasmids, with higher sensitivity than that of real-time quantitative PCR (qPCR) at 10(2) copies for the same target. Clinical samples from a total of 69 patients with probable IA (n = 14) and possible IA (n = 55) were subjected to the LAMP assay, and positive results were found for the 14 patients with probable IA (100%) and 34 patients with possible IA (61.82%). When detection using the LAMP assay was compared with that using qPCR in the 69 clinical samples, the LAMP assay demonstrated a sensitivity of 89.19% and the concordance rate for the two methods was 72.46%. Accordingly, we report that a valuable LAMP assay for the rapid, specific, and simple detection of A. fumigatus in clinical testing has been developed.

• 出版日期2016-4
• 单位中国人民解放军军事医学科学院; 四川大学