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Sun-Whan Park 1 Article
One-Step Reverse Transcription-Polymerase Chain Reaction for Ebola and Marburg Viruses
Sun-Whan Park, Ye-Ji Lee, Won-Ja Lee, Youngmee Jee, WooYoung Choi
Osong Public Health Res Perspect. 2016;7(3):205-209.   Published online June 30, 2016
DOI: https://doi.org/10.1016/j.phrp.2016.04.004
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  • 5 Citations
AbstractAbstract PDF
Objectives
Ebola and Marburg viruses (EBOVs and MARVs, respectively) are causative agents of severe hemorrhagic fever with high mortality rates in humans and nonhuman primates. In 2014, there was a major Ebola outbreak in various countries in West Africa, including Guinea, Liberia, Republic of Sierra Leone, and Nigeria. EBOV and MARV are clinically difficult to diagnose and distinguish from other African epidemic diseases. Therefore, in this study, we aimed to develop a method for rapid identification of the virus to prevent the spread of infection.
Methods
We established a conventional one-step reverse transcription-polymerase chain reaction (RT-PCR) assay for these pathogens based on the Superscript Reverse Transcriptase-Platinum Taq polymerase enzyme mixture. All assays were thoroughly optimized using in vitro-transcribed RNA.
Results
We designed seven primer sets of nucleocapsid protein (NP) genes based on sequences from seven filoviruses, including five EBOVs and two MARVs. To evaluate the sensitivity of the RT-PCR assay for each filovirus, 10-fold serial dilutions of synthetic viral RNA transcripts of EBOV or MARV NP genes were used to assess detection limits of viral RNA copies. The potential for these primers to cross react with other filoviruses was also examined. The results showed that the primers were specific for individual genotype detection in the examined filoviruses.
Conclusion
The assay established in this study may facilitate rapid, reliable laboratory diagnosis in suspected cases of Ebola and Marburg hemorrhagic fevers.

Citations

Citations to this article as recorded by  
  • Marburg Virus Disease – A Mini-Review
    Sandip Chakraborty, Deepak Chandran, Ranjan K. Mohapatra, Mahmoud Alagawany, Mohd Iqbal Yatoo, Md. Aminul Islam, Anil K. Sharma, Kuldeep Dhama
    Journal of Experimental Biology and Agricultural S.2022; 10(4): 689.     CrossRef
  • Marburgviruses: An Update
    Caterina M Miraglia
    Laboratory Medicine.2019; 50(1): 16.     CrossRef
  • Ebola virus: A global public health menace: A narrative review
    Shamimul Hasan, SyedAnsar Ahmad, Rahnuma Masood, Shazina Saeed
    Journal of Family Medicine and Primary Care.2019; 8(7): 2189.     CrossRef
  • Fast and Parallel Detection of Four Ebola Virus Species on a Microfluidic-Chip-Based Portable Reverse Transcription Loop-Mediated Isothermal Amplification System
    Xue Lin, Xiangyu Jin, Bin Xu, Ruliang Wang, Rongxin Fu, Ya Su, Kai Jiang, Han Yang, Ying Lu, Yong Guo, Guoliang Huang
    Micromachines.2019; 10(11): 777.     CrossRef
  • The current landscape of nucleic acid tests for filovirus detection
    David J. Clark, John Tyson, Andrew D. Sails, Sanjeev Krishna, Henry M. Staines
    Journal of Clinical Virology.2018; 103: 27.     CrossRef

PHRP : Osong Public Health and Research Perspectives