title: Rapid identification of Mycobacterium tuberculosis complex by a novel hybridization signal amplification method based on self-assembly of DNA-streptavidin nanoparticles.
abstract: Rapid detection of Mycobacterium tuberculosis complex (MTBC) is a critical step in controlling tuberculosis (TB). In this study, we used IS6110 as the specific identification target to develop a novel hybridization signal amplification method (HSAM) for the rapid and direct detection of MTBC from clinical sputum specimens. This system consists of magnetic bead-linked capture probes for target isolation, dextran-based nanoparticles for amplifying the reporter molecule (biotinylated-FITC), and detection probes (2B-DNA) for binding the nanoparticles. Both the capture and detection probes were specific to the IS6110 target sequence. Our results determined that as few as 10 copies of the IS6110 sequence or 10 M. tuberculosis bacteria could be detected, indicating that the HSAM assay is as sensitive as conventional PCR, and the assay was specific enough to distinguish MTBC from nontuberculosis mycobacteria (NTM). A total of 176 clinical sputum specimens were collected for HSAM evaluation, and the results were compared to those from traditional culture and biochemical identification techniques. This assay had a sensitivity of 88.3%, a specificity of 91.8%, a positive predictive value of 93.8% and a negative predictive value of 84.8% for the detection of MTBC. This technique is highly sensitive and specific, is easy to perform, and does not require any sophisticated detection equipment; thus, this approach has great potential in clinical TB detection and diagnostic applications.
authors.affiliation: Department of Pathogenobiology, Daqing Branch of Harbin Medical University , Daqing , China.
authors.forename: Haihe
authors.initials: H
authors.lastname: Wang
authors.forename: Chunyan
authors.initials: C
authors.lastname: Zhao
authors.forename: Fan
authors.initials: F
authors.lastname: Li
keywords: Detection; Diagnosis; Identification; Mycobacterium tuberculosis; Rapid methods; complex (MTBC)
references.citation: J Clin Microbiol. 1996 Dec;34(12):2985-9
references.pmid: 8940435
references.citation: Biochem Biophys Res Commun. 2006 Sep 8;347(4):1151-7
references.pmid: 16870140
references.citation: J Med Microbiol. 2005 Sep;54(Pt 9):873-878
references.pmid: 16091440
references.citation: Immunol Lett. 2006 Feb 28;103(1):27-32
references.pmid: 16325268
references.citation: J Clin Microbiol. 2000 May;38(5):1772-6
references.pmid: 10790097
references.citation: Clin Infect Dis. 2003 Jan 1;36(1):16-23
references.pmid: 12491196
references.citation: J Clin Microbiol. 1993 Jan;31(1):61-5
references.pmid: 8417034
references.citation: J Clin Microbiol. 1991 Jul;29(7):1372-5
references.pmid: 1909344
references.citation: J Clin Microbiol. 2000 Sep;38(9):3150-5
references.pmid: 10970348
references.citation: J Virol Methods. 2006 Sep;136(1-2):160-5
references.pmid: 16781785
references.citation: Diagn Microbiol Infect Dis. 2004 Apr;48(4):271-5
references.pmid: 15062920
references.citation: Braz J Microbiol. 2008 Oct;39(4):613-8
references.pmid: 24031276
references.citation: Lancet Infect Dis. 2006 Oct;6(10):664-74
references.pmid: 17008175
references.citation: J Clin Microbiol. 2002 Jul;40(7):2339-45
references.pmid: 12089245
references.citation: J Clin Microbiol. 2003 Jun;41(6):2605-15
references.pmid: 12791887
references.citation: Nucleic Acids Res. 1999 Dec 1;27(23):4553-61
references.pmid: 10556310
references.citation: Tuber Lung Dis. 1996 Aug;77(4):353-7
references.pmid: 8796252
references.citation: Clin Lab Sci. 2001 Spring;14(2):85-8
references.pmid: 15625980
references.citation: Nucleic Acids Res. 2003 Aug 15;31(16):e90
references.pmid: 12907742
references.citation: J Clin Microbiol. 2005 Dec;43(12):6086-90
references.pmid: 16333102
references.citation: Biochem Biophys Res Commun. 2007 Jun 1;357(2):391-6
references.pmid: 17428442
references.citation: J Microbiol Methods. 2006 Sep;66(3):440-8
references.pmid: 16516314
references.citation: Braz J Microbiol. 2008 Oct;39(4):673-5
references.pmid: 24031287
references.citation: Diagn Microbiol Infect Dis. 2003 Jul;46(3):211-5
references.pmid: 12867097