title: Direct and simultaneous identification of Mycobacterium tuberculosis complex (MTBC) and Mycobacterium tuberculosis (MTB) by rapid multiplex nested PCR-ICT assay.
abstract: The Mycobacterium tuberculosis (MTB) shows different virulence and host infection range from other members of the M. tuberculosis complex (MTBC). Differential identification of MTB from MTBC is thus important in certain occasions. The currently commercially available molecular assays which use either IS6110 or 16S rDNA fragment as identification targets are mainly designed for identifying MTBC but not for MTB. Comparative genomic DNA analysis has provided valuable information on regions of difference (RD) present in MTB but not in other members of the MTBC. RD9 region is further suggested to be a potential target for differential identification of MTB from MTBC. In this study, using IS6110 and Rv3618 (belong to RD9) as the specific identification targets for MTBC and MTB, respectively, we developed and tested a multiplex nested PCR-ICT (immuno-chromatography test) assay for simultaneously and directly detecting not only MTBC but also MTB from 1500 clinical sputum specimens. The results were compared with traditional culture and biochemical identification results together with patients' clinical assessments. This assay showed a 95.5% sensitivity, 97.9% specificity, 2.1% false positive rate and 4.5% false negative rate towards detection of MTBC, and a 93.0% sensitivity, 99.8% specificity, 0.2% false positive rate and 7.0% false negative rate for detection of MTB. This detection system shows great potential in clinical application.
authors.affiliation: Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University College of Medicine, No. 1 Chan-Der Street, Taipei 100, Taiwan, ROC.
authors.forename: Po-Chi
authors.initials: PC
authors.lastname: Soo
authors.forename: Yu-Tze
authors.initials: YT
authors.lastname: Horng
authors.forename: Po-Ren
authors.initials: PR
authors.lastname: Hsueh
authors.forename: Bin-Jon
authors.initials: BJ
authors.lastname: Shen
authors.forename: Jann-Yuan
authors.initials: JY
authors.lastname: Wang
authors.forename: Hui-Hsin
authors.initials: HH
authors.lastname: Tu
authors.forename: Jun-Rong
authors.initials: JR
authors.lastname: Wei
authors.forename: Shang-Chen
authors.initials: SC
authors.lastname: Hsieh
authors.forename: Chien-Chung
authors.initials: CC
authors.lastname: Huang
authors.forename: Hsin-Chih
authors.initials: HC
authors.lastname: Lai
mesh_terms: D002845:Chromatography / Q000295:instrumentation*; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005188:False Negative Reactions; D005189:False Positive Reactions; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012016:Reference Values; D012680:Sensitivity and Specificity; D014376:Tuberculosis / Q000382:microbiology*
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial