Molecular Biology Reports

, Volume 40, Issue 2, pp 1883–1892 | Cite as

A dual molecular beacon approach for fast detection of Mycobacterium tuberculosis

Article
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Abstract

The main objectives of this study were to assess a dual molecular beacon approach for fast detection of Mycobacterium tuberculosis (MT). MT beacon (Tb-B) was designed to target the unique IS6110 (114 bp) and rpoB (215 bp) fragment of the MT (H37Ra) genome, and the two fragments were inserted into the PMD-19T vector after purification, by PCR and sequencing, to construct plasmids. Different dilutions of positive plasmid standards were used for dual molecular beacon RT-PCR of rpoB and IS6110, and standard curves were established.The results show that the dual molecular beacon of rpoB and IS6110 detecting MT was stable (CV is 1.91–2.68 %) with a high amplification efficiency (95.6 %). In addition, the strains of non MT did not generate fluorescence signals, while strains of MT did, indicating that the primers and molecular beacons were specific, and only MT complex was amplified. The linear range was wide (103–1011 copies/mL), and clinical specimens presenting different bacterial counts can be detected.

Keywords

Mycobacterium tuberculosis Molecular beacons Fluorescence quantitative PCR IS6110 rpoB 

Abbreviations

MT

Mycobacterium tuberculosis

MTC

Mycobacterium tuberculosis complex

SD

Standard deviation

CV

Coefficient of variation

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Notes

Acknowledgments

This work was supported by grants from the Science Research Foundation of Health Ministry - Fujian Health Education Jointly Project, No. WKJ 2005-2-007.

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Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Chuan-Xing Yu
    • 2
  • Zi-Yun Zhao
    • 1
  • Jian-Xin Lv
    • 3
  • Ling Zhu
    • 1
  1. 1.Immunology Department and Center of NeuroscienceFujian Medical UniversityFuzhouPeople’s Republic of China
  2. 2.Internal Medicine of Second People’s Hospital of Fujian ProvinceFuzhouPeople’s Republic of China
  3. 3.Zhejiang Provincial Key Laboratory of Medical GeneticsWenzhou Medical CollegeWenzhouPeople’s Republic of China

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