title: An efficient alternative marker for specific identification of Mycobacterium tuberculosis.
abstract: Rapid and accurate identification of mycobacteria to the species level is important to provide epidemiological information and to guide the appropriate treatment, especially identification of the Mycobacterium tuberculosis (MTB) which is the leading pathogen causing tuberculosis. The genetic marker named as Mycobacterium tuberculosis specific sequence 90 (mtss90) was screened by a bioinformatics software and verified by a series of experiments. To test its specificity, 266 strains of microorganisms and human cells were used for the mtss90 conventional PCR method. Moreover, the efficiency of mtss90 was evaluated by comparing 16S rDNA (Mycobacterium genus-specific), IS6110 (specific identification of MTB complex), mtp40 (MTB-specific) and PNB/TCH method (traditional bacteriology testing) in Mycobacterium strains. All MTB isolates were mtss90 positive. No amplification was observed from any other tested strains with M. microti as an exception. Compared with the traditional PNB/TCH method, the coincidence rate was 99.1 % (233/235). All of the mtss90 positive strains were IS6110 and 16S rDNA positive, indicating a 100 % coincidence rate (216/216) between mtss90 and these two genetic markers. Additionally, mtss90 had a better specificity than mtp40 in the identification of MTB. Lastly, a real-time PCR diagnostic assay was developed for the rapid identification of MTB. In conclusion, mtss90 may be an efficient alternative marker for species-specific identification of MTB and could be used for the diagnosis of tuberculosis combined with other genetic markers.
authors.affiliation: Department of Laboratory Medicine, College of Laboratory Medicine, Chongqing Medical University, 1 Yixueyuan Road, Yuzhong District, Chongqing, 400016, People's Republic of China.
authors.forename: Jianing
authors.initials: J
authors.lastname: Zhao
authors.forename: Yiwei
authors.initials: Y
authors.lastname: Wang
authors.forename: Dairong
authors.initials: D
authors.lastname: Li
authors.forename: Jiawen
authors.initials: J
authors.lastname: Liu
authors.forename: Xuemei
authors.initials: X
authors.lastname: Zhang
authors.forename: Yujuan
authors.initials: Y
authors.lastname: He
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Ju
authors.initials: J
authors.lastname: Cao
authors.forename: Yibing
authors.initials: Y
authors.lastname: Yin
authors.forename: Wenchun
authors.initials: W
authors.lastname: Xu
mesh_terms: D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods*; D015415:Biomarkers / Q000032:analysis*; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology; D019295:Computational Biology; D004269:DNA, Bacterial / Q000032:analysis*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D009181:Mycoses / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014376:Tuberculosis / Q000382:microbiology / Q000662:veterinary
chemical_list: D015415:Biomarkers; D004269:DNA, Bacterial