COMPARATIVE STUDY OF LOOP MEDIATED ISOTHERMAL AMPLIFICATION ASSAY AND REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION TECHNIQUE IN THE DIAGNOSIS OF EBOLA VIRUS IN HUMAN BLOOD SAMPLES IN LAGOS, NIGERIA

Authors

  • L. J. OJOMO
  • F. ONIKEPE
  • J. J. AJAKAIYE
  • J. P. EROMON
  • B. BANKOLE
  • M. OGBOLE

DOI:

https://doi.org/10.33003/jaat.2021.0702.046

Abstract

The not too distant past outbreak of Ebola viral disease (EVD) in West African countries was reported to be the worst since its emergence in 1976. A major challenge to the management and control of EVD is lack of rapid and accurate diagnosis. Therefore, this study was conducted to compare the Loop Mediated Isothermal Amplification (LAMP) assay to the Reverse transcription-polymerase chain reaction (RT-PCR) technique for the diagnosis of Ebola virus (EBV) in human blood samples. EBV was confirmed in 5 (33%) and 16(12%) human blood samples by LAMP and RT-PCR, respectively. The sensitivity of LAMP assay was 83.3% while the specificity was 100% for LAMP when compared to RT-PCR. Findings from this study indicate that LAMP may be a rapid diagnostic tool that requires minimal expertise in developing countries in the diagnosis of this disease, which compares favourably to the gold standard RT-PCR. Findings from this study indicate that LAMP with necessary diagnostic improvements may be a rapid diagnostic tool that requires minimal expertise in developing countries in the diagnosis of this disease, if it is to compare favourably to the gold standard RT-PCR.

References

Charles, N. H. (2014). On the Quarantine Period for Ebola Virus. Public Library of Science Currents, 5: 121-135 doi: 10.1371/currents.outbreaks.2ab4b76ba7263ff0f084766e43abbd88.

Dhama, K., Karthik, K., Chakraborty, S., Tiwria, R., Kapoor, S., Kumar, A. and Thomas, P. (2014). Loop-mediated isothermal amplification of DNA (LAMP): a new diagnostic tool lights the world of diagnosis of animal and human pathogens: a review. Pakistan Journal of Biological Science, 17: 151-166.

Dorota, D. and Milena, K. (2013) Loop-mediated Isothermal Amplification (LAMP) as a diagnostic tool in detection of infectious diseases Microbiology Department, Faculty of Chemistry, PhD Interdisciplinary Journal 11(12): 80-233.

Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F.R. and

Smith, T. F. (2006). Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clinical Microbiology Review, 19:165-256.

Fu, S., Qu, G., Guo, S., Ma, L., Zhang, N. and Zhang, S. (2011). Applications of loop-mediated isothermal DNA amplification. Applied Biochemistry and Biotechnology, 163: 845–850.

Haradhan, K. M. (2014). The most fatal 2014 outbreak of Ebola Virus disease in West Africa. American Journal of Epidemiology and Infectious Disease, 2(4): 101-108. Doi 10.12691/ajeid-2-4-4

Kilgore, P. E., Grabenstein, J. D. and Salim, A. M. (2015). Treatment of Ebola virus disease. Pharmacotherapy, 35:43-53.

Kuhn, J. H., Becker, S., Ebihara, H., Geisbert, T. W., Johnson, K., Kawaoka, M. Y., Lipkin, W. I. and Negredo, A. I. (2010). Proposal for a revised taxonomy of the family. Filoviridae: Classification, names of taxa and viruses, and virus abbreviations. Archive of Virology, 155: 2083–2103.

Leroy, E. M., Kumulungui, B., Pourrut, X., Rouquet, P., Hassanin, A., Yaba, P., Délicat, A., Paweska, J. T., Gonzalez, J. P. and Swanepoel, R. (2005). Fruit bats as reservoirs of Ebola virus. Ebola hemorrhagic fever outbreaks in Gabon, 1994-1997. Journal of Infectious Disease, Journal of Infectious Disease, 438: 575–576.

Luc, B. and Michel, G. B. (2010). Diagnosing Infections-Current and anticipated technologies for point-of-care diagnostics and home-based testing. Clinical Microbiology and Infection, 16: 1044-1053.

Mori, Y. and Notomi, T. (2009). Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. Journal of Infectious Chemotherapy, 15: 62–69.

Ndao, M. (2009). Diagnosis of Parasitic Diseases: Old and New Approaches. Interdisciplinary Perspectives on Infectious Diseases, Special Edition, Available at URL https://doi.org/10.1155/2009/278246

Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K. and Amino, N. (2000). Loop-mediated isothermal amplification of DNA. Nucleic Acids Research, 28: 63-68.

Parida, M. S., Sannarangaiah, P. K., Dash, P. V. and Rao, K. (2008). Loop mediated isothermal amplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases. Review in Medical Virology, 18: 407–421.

Ratcliff, R. M., Chang, G., Kok, T. and Sloots, T. P. (2007). Molecular diagnosis of medical viruses. Current Issues in Molecular Biology, 9: 87–102.

Silvia, D., Rosa, C., James, S. S., Hilde, D., Grazia, M. C., Anja, W., Kathryn, S., Gabriela, P., Raphael, B., Osman, B., Armand, S., Michel, V. H. and Rony, Z. (2015). Ebola outbreak in rural West Africa: epidemiology, clinical features and outcome. Tropical Medicine and International Health, 20: 448–454.

Towner, J. S., Pierre, E., Rollin, D. G. B., Anthony, S., Sharon, M. C., Martin, V., William, F. L., Christina, F., Spiropoulou, T. G., Ksiazek, M. L., Felix, K., Robert, D. and Stuart, T. N. (2004). Rapid Diagnosis of Ebola Hemorrhagic Fever by Reverse Transcription-PCR in an Outbreak Setting and Assessment of Patient Viral Load as a Predictor of Outcome. Journal of Virology, 78(8): 4330-4341.

Tsugunori, N., Hiroto, O.,Harrumi, M.,Toshihiro, Y., Keiko, W., ,Nobuyuki, A. and Tetsu, H. (2000). Loop-mediated isothermal amplification of DNA Nucleic Acids Res. Clinical Microbiology of Infectious Disease, 15:28(12).63-69.

Tsugunori, N., Yasuyoshi, M., Norihiro, T. and Hidetoshi, K. (2015) Loop-mediated isothermal amplification (LAMP): principle, features, and future prospects. Journal of Microbiology, 53: 1-5.

WHO. (2010). WHO Recommendations on the diagnosis of HIV infection in infants and children:http://apps.who.int/iris/bitstream/handle/10665/44275/9789241599085_eng_Annexes.pdf, 2010 Accessed on 25th May, 2021.

WHO. (2015). World Health Organisation. Ebola Virus Disease: Factsheet http://www.who.int/mediacentre/factsheets/fs103/en/.2015. Accessed on 25th May, 2021.

Yasuyoshi, M. and Tsugunori, N. (2009) Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. Journal of infection and chemotherapy. 15(2): 62-69.

Yasuyoshi, M. and Tsugunori, N. (2013). Loop-mediated isothermal amplification (LAMP): Recent progress in research and development Journal of Infection and Chemotherapy,19: 401-411.

Yogesh, C., Koelbl, J., Puckett, J., Moser, M. J., Klingele, A. J., Liles, M. R., Carrias, A., Mead, D. A. and Schoenfeld, T. W. (2014). A novel thermostable polymerase for RNA and DNA loop-mediated isothermal amplification (LAMP). Frontiers in Microbiology 5:395-408.

Downloads

Published

2022-06-06