Organizing co mmittee

 
 

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Plenary presentation
3 
SARS-COV-2: NEW DEVELOPMENTS FOR PREVENTION AND 
DIAGNOSIS 
Sh. Azimova, S. Sasmakov, F. Eshboev, J. Abdurakhmanov, O. Ashirov, Sh. Khasanov, 
A. Makhnyov, Kh. Nasriddinov, A. Boymirzaev, M. Umarova, E. Yusupova, G. Piyakina, 
U. Khamidova, S. Gaynazarova, T. Sadullaev, A. Yarilkaganova, S. Ikramov, Kh. 
Dolimov, O. Alimukhamedova, N. Tosheva, D. Mansurov, E. Lysova, E Terenteva. 
Acad. S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences of 
the Republic of Uzbekistan 
SARS-CoV-2 (severe acute respiratory syndrome-related coronavirus 2) is a single-stranded 
(+) RNA containing virus. Currently used vaccines mainly targets the receptor-binding domain 
(RBD) of the surface S (spike) protein of virus. However, the largest number of mutations is 
found in the spike protein of the coronavirus, for example, in the last widespread strain Omicron 
(SARS-CoV-2 B.1.1.529), there are more than 30 of mutations. It was established that N protein 
(Nucleocapsid) of SARS-CoV-2 is highly conserved and antigenic regions of the N protein are 
recognized by T cells on the surface of infected cells, which indicates the role of the N protein 
in generating not only the primary humoral (B), but also the cellular (T) immune response to 
infection of SARS-CoV-2. In this regard, the SARS-CoV-2 Nucleocapsid (N) or its 
combination with the S protein may be the most suitable candidate for the development of 
effective vaccines.
For developing vaccine to the SARS-CoV-2 we have used different regions of Spike. 14 new 
recombinant plasmids encoding S proteins (Spike1259, Spike906, Spike1020, Spike2136, 
Spike3820), Omicron-RBD, and nucleocapsid (N) were cloned for expression systems 
Pichia 
pastoris
and 
Bombyx mori 
baculoviruses/insect cells. 
The effective expression of recombinant Spike proteins (subunits), Omicron-RBD and 
nucleocapsid (N) in 
Pichia pastoris
yeast cells were confirmed by PCR, ELISA, and 
Immunoblotting. The optimal conditions for cultivating of recombinant strains of 
Pichia 
pastoris
were selected. 
Recombinant baculoviruses expressing target SARS-CoV-2 genes (Spike 1259, 906, 2136 
and Nucelocapsid N) were obtained on the basis of nuclear polyhedrosis virus of 
Bombyx mori
(BmNPV). Synthesis of target recombinant proteins in larvae (silk worm) was determined by 
ELISA.
Methods for isolation and purifying of target recombinant proteins have been developed. 
The results of the study on immunogenicity in laboratory animals (mice) for recombinant 
proteins Omicron-RBD, Nucleocapsid N SARS-CoV-2 and their combination showed high 
efficiency and safety. 
For the prevention of SARS-CoV-2 infection were developed new drugs - combinations 
based on RNase and RNase/Niclosamide included in a liposome. The safety of developed drug 
confirmed by the studies on laboratory animals. According to the results of studies, the 
effectiveness of the drug in reducing the viral load by more than 1000 times it was established. 
For determination of SARS-CoV-2 coronavirus RNA was developed a PCR diagnostic kit 
using hybridization-fluorescence detection. The sensitivity of PCR kit is at least 50 copies of 
coronavirus RNA in a PCR sample. The specificity of the PCR assay is 100%. Moreover, a set 
of reagents "ENCOR" for the extraction of nucleic acids (DNA/RNA) from biological samples 
such as blood plasma, swabs and scrapings of human mucous membranes in order to diagnose 
infectious diseases (SARS-CoV-2, HBV, HCV, HIV, TORCH, and etc.) was designed. The 
“SARS-CoV-2 PCR DETECT” diagnostic kit and set of reagents "ENCOR" were registered 
with the Ministry of Health of the Republic of Uzbekistan (Registration certificates TB/M 
00585/08/22 and TB/M 00397/06/20).

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Plenary presentation 
4