Whole Genome Sequencing of SARS-CoV-2: Adapting Illumina Protocols for Quick and Accurate Outbreak Investigation during a Pandemic

Genes

Yunus Moosa¹  Houriiyah Tegally²  Richard Lessells²  Sureshnee Pillay²  Eduan Wilkinson²  Inbal Gazy²  Lavanya Singh²  Jennifer Giandhari²  JamesEmmanuel San²  Stacey Mattison²  Vagner Fonseca²  Maryam Fish²  Tulio de Oliveira²  KhulekaniSedwell Khanyile²  Benjamin Chimukangara²  LuizCarlos Jr. Alcantara³  Marta Giovanetti⁴ 

[1] Infectious Diseases Department, Nelson R Mandela School of Medicine, University of KwaZulu-Natal, Durban 4001, South Africa;KwaZulu-Natal Research Innovation and Sequencing Platform (KRISP), School of Laboratory Medicine & Medical Sciences, University of KwaZulu-Natal, Durban 4001, South Africa;Laboratorio de Genetica Celular e Molecular, ICB, Universidade Federal de Minas Gerais, Belo Horizonte 31270-901, Brazil;Laboratório de Flavivírus, Instituto Oswaldo Cruz Fiocruz, Rio de Janeiro 21045-900, Brazil;

关键词: sequencing;    SARS-CoV2;    Illumina;    protocols;    COVID-19;    bioinformatics;   

DOI  :  10.3390/genes11080949

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown