Article
Whole Genome Sequencing of SARS-CoV-2: Adapting Illumina Protocols for Quick and Accurate Outbreak Investigation during a Pandemic
Registro en:
PILLAY, Sureshnne; et al. Whole Genome Sequencing of SARS-CoV-2: Adapting Illumina Protocols for Quick and Accurate Outbreak Investigation during a Pandemic. Genes, v. 11, n. 949, p. 1-13, 2020.
2073-4425
10.3390/genes11080949
Autor
Pillay, Sureshnee
Giandhari, Jennifer
Tegally, Houriiyah
Wilkinson, Eduan
Chimukangara, Benjamin
Lessells, Richard
Moosa, Yunus
Mattison, Stacey
Gazy, Inbal
Fish, Maryam
Singh, Lavanya
Khanyile, Khulekani Sedwell
San, James Emmanuel
Fonseca, Vagner
Giovanetti, Marta
Alcantara, Luiz Carlos
Oliveira, Tulio de
Resumen
The COVID-19 pandemic has spread very fast around the world. A few days after the first detected case in South Africa, an infection started in a large hospital outbreak in Durban, KwaZulu-Natal (KZN). Phylogenetic analysis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genomes can be used to trace the path of transmission within a hospital. It can also identify the source of the outbreak and provide lessons to improve infection prevention and control strategies. This manuscript outlines the obstacles encountered in order to genotype SARS-CoV-2 in near-real time during an urgent outbreak investigation. This included problems with the length of the original genotyping protocol, unavailability of reagents, and sample degradation and storage. Despite this, three different library preparation methods for Illumina sequencing were set up, and the hands-on library preparation time was decreased from twelve to three hours, which enabled the outbreak investigation to be completed in just a few weeks. Furthermore, the new protocols increased the success rate of sequencing whole viral genomes. A simple bioinformatics workflow for the assembly of high-quality genomes in near-real time was also fine-tuned. In order to allow other laboratories to learn from our experience, all of the library preparation and bioinformatics protocols are publicly available at protocols.io and distributed to other laboratories of the Network for Genomics Surveillance in South Africa (NGS-SA) consortium.