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2020 ; 84
(ä): 104390
Nephropedia Template TP
gab.com Text
Twit Text FOAVip
Twit Text #
English Wikipedia
Molecular analysis of several in-house rRT-PCR protocols for SARS-CoV-2 detection
in the context of genetic variability of the virus in Colombia
#MMPMID32505692
Álvarez-Díaz DA
; Franco-Muñoz C
; Laiton-Donato K
; Usme-Ciro JA
; Franco-Sierra ND
; Flórez-Sánchez AC
; Gómez-Rangel S
; Rodríguez-Calderon LD
; Barbosa-Ramirez J
; Ospitia-Baez E
; Walteros DM
; Ospina-Martinez ML
; Mercado-Reyes M
Infect Genet Evol
2020[Oct]; 84
(ä): 104390
PMID32505692
show ga
The COVID-19 pandemic caused by SARS-CoV-2 is a public health problem
unprecedented in the recent history of humanity. Different in-house real-time
RT-PCR (rRT-PCR) methods for SARS-CoV-2 diagnosis and the appearance of genomes
with mutations in primer regions have been reported. Hence, whole-genome data
from locally-circulating SARS-CoV-2 strains contribute to the knowledge of its
global variability and the development and fine tuning of diagnostic protocols.
To describe the genetic variability of Colombian SARS-CoV-2 genomes in
hybridization regions of oligonucleotides of the main in-house methods for
SARS-CoV-2 detection, RNA samples with confirmed SARS-CoV-2 molecular diagnosis
were processed through next-generation sequencing. Primers/probes sequences from
13 target regions for SARS-CoV-2 detection suggested by 7 institutions and
consolidated by WHO during the early stage of the pandemic were aligned with
Muscle tool to assess the genetic variability potentially affecting their
performance. Finally, the corresponding codon positions at the 3' end of each
primer, the open reading frame inspection was identified for each gene/protein
product. Complete SARS-CoV-2 genomes were obtained from 30 COVID-19 cases,
representative of the current epidemiology in the country. Mismatches between at
least one Colombian sequence and five oligonucleotides targeting the RdRP and N
genes were observed. The 3' end of 4 primers aligned to the third codon position,
showed high risk of nucleotide substitution and potential mismatches at this
critical position. Genetic variability was detected in Colombian SARS-CoV-2
sequences in some of the primer/probe regions for in-house rRT-PCR diagnostic
tests available at WHO COVID-19 technical guidelines; its impact on the
performance and rates of false-negative results should be experimentally
evaluated. The genomic surveillance of SARS-CoV-2 is highly recommended for the
early identification of mutations in critical regions and to issue
recommendations on specific diagnostic tests to ensure the coverage of
locally-circulating genetic variants.