Development and evaluation of a one-step quantitative RT-PCR assay for detection of Lassa virus

Dedkov V.G., Magassouba N.F., Safonova M.V., Naydenova E.V., Ayginin A.A., Kourouma B.C., Camara A.B., Camara J., Kritzkiy A.A., Tuchkov I.V., Shchelkanov M.Yu., Maleev V.V.

В журнале Journal of Virological Methods

Год: 2019 Том: 271 ArticleID: 113674

Lassa fever is a severe viral hemorrhagic illness caused by Lassa virus. Based on estimates, the number of LASV infections ranges from 300,000 to 500,000 cases in endemic areas witha fatality rate of1%. Development of fast and sensitive tools for the control and prevention of Lassa virus infection as well as for clinical diagnostics of Lassa fever are crucial. Here we reported development and evaluation of a one-step quantitative RT-qPCR assay for the Lassa virus detection – LASV-Fl. This assay is suitable for the detection of lineages I-IV of Lassa virus. The limit of detection of the assay ranged from 103 copies/ml to 105 copies/ml and has 96.4% diagnostic sensitivity, whereas analytical and diagnostic speci?cities both were 100%. Serum, whole blood and tissue are suitable for use with the assay. The assay contains all the necessary components to perform the analysis, including an armored positive control (ARC+)andan armoredinternalcontrol(IC). Thestudy was done duringthe missionofspecialized antiepidemic team of the Russian Federation (SAET) in the Republic of Guinea in 2015-2018. Basedonsequencingdata,LASV-speci?cassaywasdevelopedusingsyntheticMS2-phage-basedarmoredRNA particles, RNA from Lassa virus strain Josiah, and further, evaluated in ?eld conditions using samples from patients and Mastomys natalensis rodents.

DOI 10.1016/j.jviromet.2019.113674

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