title: Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Rapid Detection of Aspergillus fumigatus.
abstract: Aspergillus fumigatusis a conditional pathogen and the major cause of life-threatening invasive aspergillosis (IA) in immunocompromised patients. The early and rapid detection ofA. fumigatusinfection 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 ofA. 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. fumigatusstrains, including 5 species of theAspergillusgenus. 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 ofA. fumigatusin clinical testing has been developed.
authors.affiliation: Key Laboratory of Bio-resources and Eco-environment of the Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, Sichuan, People's Republic of China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Tang
authors.affiliation: Department for Hospital Infection Control and Research, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China.
authors.forename: Shuguang
authors.initials: S
authors.lastname: Tian
authors.affiliation: Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China.
authors.forename: Nong
authors.initials: N
authors.lastname: Yu
authors.affiliation: Department for Hospital Infection Control and Research, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department for Hospital Infection Control and Research, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Jia
authors.affiliation: Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Zhai
authors.affiliation: Key Laboratory of Bio-resources and Eco-environment of the Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, Sichuan, People's Republic of China qunsun@scu.edu.cn hanlicdc@163.com.
authors.forename: Qun
authors.initials: Q
authors.lastname: Sun
authors.affiliation: Department for Hospital Infection Control and Research, Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China qunsun@scu.edu.cn hanlicdc@163.com.
authors.forename: Li
authors.initials: L
authors.lastname: Han
mesh_terms: D017302:Annexins / Q000235:genetics; D001228:Aspergillosis / Q000175:diagnosis*; D001232:Aspergillus fumigatus / Q000302:isolation & purification*; D005656:Fungal Proteins / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D017302:Annexins; D005656:Fungal Proteins