Проблемы особо опасных инфекций (Dec 2018)

Development and Testing of the Method for the Detection of Lassa virus RNA, Based on real-Time Polymerase Chain reaction with reverse Transcription

  • V. G. Dedkov,
  • M. V. Safonova,
  • E. V. Naidenova,
  • N. F. Magassouba,
  • A. A. Ayginin,
  • B. Soropogui,
  • F. Kourouma,
  • A. B. Camara,
  • J. Camara,
  • A. A. Kritsky,
  • M. Yu. Shchelkanov,
  • V. V. Maleev

DOI
https://doi.org/10.21055/0370-1069-2018-4-39-47
Journal volume & issue
Vol. 0, no. 4
pp. 39 – 47

Abstract

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Abstract. Objective of the study was the development of a method for the detection and quantitative analysis (realtime RT-PCR) to identify genetic markers of Lassa virus - LASV-Fl. Materials and methods. We utilized all the available in the GenBank database (https://www.ncbi.nlm.nih.gov/genbank/) Lassa virus sequences that have been aligned to identify conservative sites applying the BioEdit 7.2.5 software package (IbisBiosciences, USA). To test the developed PCR kit, the control panel of Lassa virus RNA and pseudo-viral particles, 27 viral strains belonging to different fami­lies, as well as 37 serum samples from patients with feverish diseases selected in medical institutions of the Republic of Guinea in 2016-2018 and 55 samples of organ suspensions from multi-spiked mice were used. Results and discussion. The analytical sensitivity of the method varied from 103 copies/ml to 105 copies/ml and had 96.4 % diagnostic sensitivity, while the analytical and diagnostic specificity was 100 %. It is shown that the developed technique can be successfully introduced into practice for the detection of Lassa virus in the Republic of Guinea, using various types of material from small mammals, including whole blood and organ suspensions of M. natalensis, as well as samples of human blood sera collected 3-7 days after the onset of the disease. It is also suggested that this method can be used for strains of Lassa virus, common not only in Guinea but also in other endemic areas, but this fact must be confirmed in further studies.

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