title: Real-time fluorescence Loop-Mediated Isothermal Amplification (LAMP) for rapid and reliable diagnosis of pulmonary tuberculosis.
abstract: A reliable, simple and rapid diagnostic method that can be helpful in pulmonary tuberculosis diagnosis is urgently needed. Loop-mediated Isothermal Amplification (LAMP) allows DNA to be amplified rapidly at a constant temperature. In this study, real-time fluorescence LAMP was evaluated to rapidly detect Mycobacterium tuberculosis in sputum and was compared to the performance of real-time fluorescence quantitative PCR (Q-PCR). All the standard MTB strains were successfully detected and limit of detection (LOD) was 10(2)CFU/mL by real-time fluorescence LAMP within 20min. In light of MTB in sputum, the real-time fluorescence LAMP method yielded a sensitivity of 98.0% and a specificity of 78.3%, compared to Q-PCR assay, which yielded a sensitivity of 96.0% and a specificity of 82.6% for PTB diagnosis. There was an excellent overall agreement between LAMP and Q-PCR for PTB (κ=0.315) and non-PTB (κ=0.862). Therefore, the real-time fluorescence LAMP assay is a rapid, sensitive, and specific method to detect pulmonary tuberculosis.
authors.affiliation: Department of Laboratory Medicine, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China.
authors.forename: Donglin
authors.initials: D
authors.lastname: Cao
authors.affiliation: Department of Laboratory Medicine, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China.
authors.forename: Liangshan
authors.initials: L
authors.lastname: Hu
authors.affiliation: Department of Laboratory Medicine, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China.
authors.forename: Maorui
authors.initials: M
authors.lastname: Lin
authors.affiliation: Department of Laboratory Medicine, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China.
authors.forename: Mingyou
authors.initials: M
authors.lastname: Li
authors.affiliation: Department of Emergency, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China.
authors.forename: Zebing
authors.initials: Z
authors.lastname: Ye
authors.affiliation: Department of Osteology, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China.
authors.forename: Hongtao
authors.initials: H
authors.lastname: Sun
authors.affiliation: Department of Laboratory Medicine, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China.
authors.forename: Jiwei
authors.initials: J
authors.lastname: Huang
authors.affiliation: Department of Laboratory Medicine, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China.
authors.forename: Huawen
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Laboratory Medicine, Guangdong No.2 Provincial People's Hospital, Guangzhou 510317, PR China. Electronic address: caodlgz@sina.com.
authors.forename: Junzhang
authors.initials: J
authors.lastname: Tian
mesh_terms: D000328:Adult; D000368:Aged; D001431:Bacteriological Techniques / Q000379:methods*; D005260:Female; D005453:Fluorescence; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis; D055815:Young Adult
keywords: Loop-Mediated Isothermal Amplification (LAMP); Pulmonary tuberculosis; Rapid diagnosis