title: A novel method for diagnosis of smear-negative tuberculosis patients by combining a random unbiased Phi29 amplification with a specific real-time PCR.
abstract: In this study, we develop a novel method for diagnosis of smear-negative tuberculosis patients by performing a random unbiased Phi29 amplification prior to the use of a specific real-time PCR. The limit of detection (LOD) of the conventional real-time PCR was 100 colony-forming units (CFU) of MTB genome/reaction, while the REPLI real-time PCR assay could detect 0.4 CFU/reaction. In comparison with the conventional real-time PCR, REPLI real-time PCR shows better sensitivity for the detection of smear-negative tuberculosis (P = 0.015).
authors.affiliation: National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Pang
authors.affiliation: Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, Beijing, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Lu
authors.affiliation: Shaanxi Tuberculosis Dispensary, Shaanxi, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Yang
authors.affiliation: National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Wang
authors.affiliation: McGill International TB Centre, Montreal, Quebec, Canada.
authors.forename: Chad
authors.initials: C
authors.lastname: Cohen
authors.affiliation: Beijing Key Laboratory for Pediatric Diseases of Otolaryngology, Head and Neck Surgery, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, Beijing, China. Electronic address: nixin@bch.com.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Ni
authors.affiliation: National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China. Electronic address: zhaoyanlin@tb123.org.
authors.forename: Yanlin
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D001431:Bacteriological Techniques; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D006801:Humans; D057230:Limit of Detection; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D011237:Predictive Value of Tests; D011446:Prospective Studies; D060888:Real-Time Polymerase Chain Reaction; D013183:Sputum / Q000382:microbiology; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000235:genetics / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial; D004259:DNA-Directed DNA Polymerase
keywords: Diagnostics; REPLI real-time PCR; Smear negative; Tuberculosis