Development of Sensitive and Rapid RNA Transcription-based Isothermal Amplification Method for Detection of Mycobacterium tuberculosis

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RAMEZANI REIHANEH; FOROUZANDEH MOGHADAM MAHDI; RASAEE MOHAMMAD JAVAD

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Journal: Avicenna Journal of Medical Biotechnology Year:2019 | Volume:11 | Issue:2 Page(s): 169-175

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Title

Development of Sensitive and Rapid RNA Transcription-based Isothermal Amplification Method for Detection of Mycobacterium tuberculosis

Author(s)

RAMEZANI REIHANEH | FOROUZANDEH MOGHADAM MAHDI | RASAEE MOHAMMAD JAVAD | Issue Writer Certificate

Keywords

Microbial viability

Mycobacterium tuberculosis

NASBA-ELISA

Abstract

Background: The accurate and early diagnosis of tuberculosis is important for its ef-fective management. During the last decade, several molecular methods for detec-tion of Tuberculosis (TB) have been developed. Since RNA especially mRNA has a generally much shorter half-life than DNA, its detection may be useful for the assess-ment of viability of bacteria. This research is a Nucleic Acid Sequence Based Amplifi-cation-Enzyme Linked Immunosorbent Assay (NASBA-ELISA) which was designed and developed for rapid detection of viable Mycobacterium tuberculosis (M. tubercu-losis). Methods: Oligonucleotide primers targeting tuf gene encoding viability marker EF-Tu mRNAs were selected and used for the amplification of mycobacterial RNA by the isothermal NASBA Digoxigenin (DIG) labeling process and incorporated with DIG-UTP, reverse transcriptase and T7 RNA polymerase. Results: Using the NASBA-ELISA system, as little as 17. 5 pg of RNA of M. tuberculosis was detected within 4 hr and no interference was encountered in the amplification and detection of viable M. tuberculosis in the presence of non-target RNA or DNA. Results obtained from the clinical specimens showed 97 and 75% of sensitivity and specificity, respectively. Conclusion: The NASBA-ELISA system offers several advantages in terms of sensitivity, rapidity and simplicity for detection of M. tuberculosis. Furthermore, due to its simplic-ity and high sensitivity feature, it could be used in limited access laboratories in a cost-effective manner.

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APA: Copy

RAMEZANI, REIHANEH, FOROUZANDEH MOGHADAM, MAHDI, & RASAEE, MOHAMMAD JAVAD. (2019). Development of Sensitive and Rapid RNA Transcription-based Isothermal Amplification Method for Detection of Mycobacterium tuberculosis. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), 11(2), 169-175. SID. https://sid.ir/paper/314028/en

Vancouver: Copy

RAMEZANI REIHANEH, FOROUZANDEH MOGHADAM MAHDI, RASAEE MOHAMMAD JAVAD. Development of Sensitive and Rapid RNA Transcription-based Isothermal Amplification Method for Detection of Mycobacterium tuberculosis. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)[Internet]. 2019;11(2):169-175. Available from: https://sid.ir/paper/314028/en

IEEE: Copy

REIHANEH RAMEZANI, MAHDI FOROUZANDEH MOGHADAM, and MOHAMMAD JAVAD RASAEE, “Development of Sensitive and Rapid RNA Transcription-based Isothermal Amplification Method for Detection of Mycobacterium tuberculosis,” AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), vol. 11, no. 2, pp. 169–175, 2019, [Online]. Available: https://sid.ir/paper/314028/en

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