Osong Public Health and Research Perspectives

Original 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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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
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Caterina M Miraglia
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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

Review Article

Travel-Associated Chikungunya Cases in South Korea during 2009–2010: Go Woon Cha, Jung Eun Cho, Eun Ju Lee, Young Ran Ju, Myung Guk Han, Chan Park, Young Eui Jeong; Osong Public Health Res Perspect. 2013;4(3):170-175. Published online June 30, 2013; DOI: https://doi.org/10.1016/j.phrp.2013.04.008

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Abstract PDF

Objectives
Chikungunya (CHIK) has been classified as a communicable disease group IV in South Korea since late 2010. Based on this, we investigated the extent of imported cases of CHIK in dengue-suspected individuals returning from dengue-endemic regions.
Methods
A total of 486 dengue-suspected serum samples were screened for CHIK by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) analysis. Further RT-PCR-positive samples were used for the viral culture, and CHIK was subsequently confirmed by sequence analysis of the culture samples.
Results
Five out of 107 dengue-positive samples were found to be positive for CHIK and 15 out of 379 dengue-negative samples were found to be positive for CHIK by immunoglobulin M ELISA. Further, a CHIK virus was isolated from one of the two RT-PCR-positive sera by cell culture and confirmed by sequence analysis.
Conclusion
The present study documents the first evidence of travel-associated CHIK infection in South Korea. Considering the intense international traffic between countries, our finding emphasizes the urgent need for active patient and vector surveillance for timely response to reduce the introduction of CHIK in Korea.

Citations

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