title: Development of an assay for genetic analysis of different SARS-CoV-2 virus variants
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¼Ð°Ð³Ð¸ÑÑÑÐ°
titleType: Subtitle
title: Ð Ð°Ð·ÑÐ°Ð±Ð¾ÑÐºÐ° Ð¿ÑÐ¾ÑÐ¾ÐºÐ¾Ð»Ð° Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¾Ð³Ð¾ Ð°Ð½Ð°Ð»Ð¸Ð·Ð° ÐºÐ¾ÑÐ¾Ð½Ð°Ð²Ð¸ÑÑÑÐ¾Ð² SARSâCoVâ2 ÑÐ°Ð·Ð½ÑÑ Ð»Ð¸Ð½Ð¸Ð¹
Creators: name: Ð§Ð¸ÑÑÑÐºÐ¾Ð²Ð°, ÐÐ½Ð½Ð°
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2023
Subjects: ÐÐ¸ÑÑÑÐ¾Ð»Ð¾Ð³Ð¸Ñ
ÐÐ¸Ð¾ÑÐµÑ Ð½Ð¾Ð»Ð¾Ð³Ð¸Ñ
ÐºÐ¾ÑÐ¾Ð½Ð°Ð²Ð¸ÑÑÑ
ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ
Ð¿Ð¸ÑÐ¾ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ
coronavirus
sequencing
pyrosequencing

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐºÐµ Ð¿ÑÐ¾ÑÐ¾ÐºÐ¾Ð»Ð¾Ð² Ð¾Ð¿ÑÐµÐ´ÐµÐ»ÐµÐ½Ð¸Ñ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾ÑÑÐ¸ Ð¸ Ð³ÐµÐ½Ð¾ÑÐ¸Ð¿Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ ÑÐ°Ð·Ð»Ð¸ÑÐ½ÑÑ Ð²Ð°ÑÐ¸Ð°Ð½ÑÐ¾Ð² ÐºÐ¾ÑÐ¾Ð½Ð°Ð²Ð¸ÑÑÑÐ° SARSâCoVâ2. ÐÑÐ»Ð¸ ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐ°Ð½Ñ Ð¿ÑÐ¾ÑÐ¾ÐºÐ¾Ð»Ñ ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾ÑÑÐ¸ S Ð¸ N Ð³ÐµÐ½Ð¾Ð² Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ Ð¡ÑÐ½Ð³ÐµÑÐ°, Ð° ÑÐ°ÐºÐ¶Ðµ Ð¿ÑÐ¾ÑÐ¾ÐºÐ¾Ð» Ð³ÐµÐ½Ð¾ÑÐ¸Ð¿Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ñ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸ÐµÐ¼ Ð¿Ð¸ÑÐ¾ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ð½Ð° Ð¿ÑÐ¸Ð±Ð¾ÑÐµ PyroMark Q24. Ð ÑÐ¾Ð´Ðµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð±ÑÐ»Ð¸ Ð¿Ð¾Ð´Ð¾Ð±ÑÐ°Ð½Ñ Ð¿Ð°ÑÑ Ð¾Ð»Ð¸Ð³Ð¾Ð½ÑÐºÐ»ÐµÐ¾ÑÐ¸Ð´Ð¾Ð², Ð¾ÑÑÐ°Ð±Ð¾ÑÐ°Ð½Ñ ÑÑÐ»Ð¾Ð²Ð¸Ñ ÐÐ¢âÐÐ¦Ð  Ð´Ð»Ñ Ð°Ð¼Ð¿Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Ð²ÑÐ±ÑÐ°Ð½Ð½ÑÑ ÑÑÐ°Ð³Ð¼ÐµÐ½ÑÐ¾Ð² Ð³ÐµÐ½Ð¾Ð¼Ð°, Ð¿Ð¾Ð´Ð¾Ð±ÑÐ°Ð½Ñ Ð¿ÑÐ°Ð¹Ð¼ÐµÑÑ ÑÑÐ°Ð¿Ð¾Ð² ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ð¸ Ð¿Ð¸ÑÐ¾ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ, ÑÑÐ»Ð¾Ð²Ð¸Ñ Ð¿Ð¸ÑÐ¾ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ (Ð¿ÑÐ¾Ð³ÑÐ°Ð¼Ð¼Ð°, Ð¿Ð¾ÑÑÐ´Ð¾Ðº Ð²Ð½ÐµÑÐµÐ½Ð¸Ñ Ð½ÑÐºÐ»ÐµÐ¾ÑÐ¸Ð´Ð¾Ð²), Ð¸ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð° Ð°Ð¿ÑÐ¾Ð±Ð°ÑÐ¸Ñ ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐ°Ð½Ð½ÑÑ Ð¿ÑÐ¾ÑÐ¾ÐºÐ¾Ð»Ð¾Ð². ÐÐ¾Ð»ÑÑÐµÐ½Ð½ÑÐµ Ð¿ÑÐ¾ÑÐ¾ÐºÐ¾Ð»Ñ Ð¿Ð¾Ð·Ð²Ð¾Ð»ÑÑÑ Ð¾Ð¿ÑÐµÐ´ÐµÐ»Ð¸ÑÑ Ð¿Ð¾Ð»Ð½ÑÑ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾ÑÑÑ S Ð¸ N Ð³ÐµÐ½Ð¾Ð² Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ Ð¡ÑÐ½Ð³ÐµÑÐ°, Ð° ÑÐ°ÐºÐ¶Ðµ ÑÐ°ÑÑÐ¸ÑÑÐ¾Ð²ÑÐ²Ð°ÑÑ Ð¿Ð¸ÑÐ¾ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸ÐµÐ¼ ÑÑÐ°ÑÑÐºÐ¸ Ð¿Ð¾ÑÐ»ÐµÐ´Ð¾Ð²Ð°ÑÐµÐ»ÑÐ½Ð¾ÑÑÐ¸ S Ð³ÐµÐ½Ð°, ÐºÐ¾Ð´Ð¸ÑÑÑÑÐ¸Ðµ Ð·Ð°Ð¼ÐµÐ½Ñ, ÑÐ°ÑÐ°ÐºÑÐµÑÐ½ÑÐµ Ð´Ð»Ñ ÑÐ°Ð·Ð½ÑÑ Ð²Ð°ÑÐ¸Ð°Ð½ÑÐ¾Ð² SARSâCoVâ2: L18F, T19R, T20N, A67V, Î69â70, G142D, V213G, D405N, K417N/T, L452R, S477N, T478K, E484A/K/Q, H655Y Ð¸ Ð´Ñ. Ð Ð°Ð·ÑÐ°Ð±Ð¾ÑÐ°Ð½Ð½Ð°Ñ ÑÐ¸ÑÑÐµÐ¼Ð° Ð´ÐµÑÐµÐºÑÐ¸Ð¸ Ð¿ÑÐ¸Ð¼ÐµÐ½ÑÐµÑÑÑ Ð´Ð»Ñ ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ð¸ Ð³ÐµÐ½Ð¾ÑÐ¸Ð¿Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ ÐºÐ¾ÑÐ¾Ð½Ð°Ð²Ð¸ÑÑÑÐ¾Ð² SARSâCoVâ2 Ð¿ÑÐ¸ Ð²ÑÐ¿Ð¾Ð»Ð½ÐµÐ½Ð¸Ð¸ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ð¹ Ð² Ð¾ÑÐ´ÐµÐ»Ðµ Ð²Ð¸ÑÑÑÐ¾Ð»Ð¾Ð³Ð¸Ð¸ Ð¸Ð½ÑÑÐ¸ÑÑÑÐ° ÑÐºÑÐ¿ÐµÑÐ¸Ð¼ÐµÐ½ÑÐ°Ð»ÑÐ½Ð¾Ð¹ Ð¼ÐµÐ´Ð¸ÑÐ¸Ð½Ñ.
description: This study is devoted to the development of protocols for sequencing and genotyping of different variants of the SARSâCoVâ2 coronavirus. The developed protocols allow to determine the fullâlength sequence of S and N genes of SARSâCoVâ2 coronovirus by Sanger method and to perform genotyping of the viruses by pyrosequencing with PyroMark Q24 instrument. As a result, pairs of oligonucleotides were selected, reverse transcription PCR conditions for amplification of selected genome fragments were defined, the oligonucleotides for sequencing and pyrosequencing were developed, the pyrosequencing conditions (protocol for the instrument, dispensation order for nucleotides) were set. All developed protocols were tested with genetic material of actual coronavirus strains. The obtained assays provide sequencing of S and N genes, and genotyping protocols allow to perform pyrosequencing of S gene fragments coding numerous substitutions characterizing SARSâCoVâ2 variants: L18F, T19R, T20N, A67V, Î69â70, G142D, V213G, D405N, K417N/T, L452R, S477N, T478K, E484A/K/Q, H655Y, etc. The developed protocols are used for sequencing and genotyping of SARSâCoVâ2 strains in the Department of Virology of the Institute of Experimental Medicine.