Multi-centre collaborative rapid-response formulation and validation of a lysis buffer for extraction of SARS-CoV-2 RNA for Covid-19 testing

Martina Scallan
University College Cork

Catherine Dempsey
Cork University Hospital

John MacSharry
University College Cork

Isabelle O’Callaghan
Cork University Hospital

Abstract
Authors:
Martina F. Scallan*^1, Catherine Dempsey*^2, John MacSharry1,3,4, Isabelle O’Callaghan2, Paula M. O’Connor4,5, Conor P. Horgan6, Edel Durack7, Paul D. Cotter4,5, Sarah Hudson8, Humphrey A. Moynihan6 & Brigid Lucey,9,10
1School of Microbiology, University College Cork, College Road, Cork, Ireland; 2Department of Clinical Microbiology, Cork University Hospital, Wilton, Cork, Ireland; 3School of Medicine, UCC; 4APC Microbiome Ireland, UCC; 5Teagasc Food Research Centre, Moorepark, Co. Cork, Ireland; 6Eli Lilly Kinsale Ltd, Kinsale, Co. Cork, Ireland; 7Bernal Institute, University of Limerick, Limerick, Ireland; 8Department of Chemical Sciences, University of Limerick; Ireland 9Department of Biological Sciences, Cork Institute of Technology, Bishopstown, Cork, Ireland; 10Academy of Clinical Science & Laboratory Medicine, Old Kilmainham, Dublin 8. Ireland

The global demand for reagents for real-time reverse transcriptase PCR (RT-PCR) diagnostic tests for COVID-19 has caused a bottle neck across Ireland and the UK in efforts to follow the World Health Organisation’s advice to “Test, Test, Test”. Normally, hospital laboratories buy in reagents that allow them to concentrate on the business of diagnosis of patients and the preparation of their own reagents is rare. However, an exponential increase in demand has resulted in a shortage of numerous reagents, not least the lysis buffer required to extract the viral RNA.
In straitened times, society needs to pool its resources and expertise to best impact. This case study and validation represents an immediate and self-selected response from the scientific community, following a call by the Academy of Clinical and Laboratory Medicine, which is the professional body for medical scientists in Ireland.
Here, we describe a rapid collective effort in Ireland by medical laboratory scientists, academic researchers and the biopharma industry to generate a validated lysis buffer. We have formulated a 4M Guanidinium thiocyanate (GITC)/ Triton X-100 Lysis buffer which provides comparable results with the recommended reagents. This buffer will ease the burden on hospital labs in their heroic efforts to diagnose a large population of patients.
The purpose of this case study was to show how, in an urgent situation, a group of 11 scientists have worked collectively from five centres to pool expertise, technical know-how, chemicals from their research laboratories, their facilities, time, co-ordination and communication skills, along with determination to develop a suitable replacement lysis buffer at scale and a validation paper in less than two weeks. Four formulations were developed in two research centres, a risk assessment for the preparation of buffers was prepared in a separate centre, validation of each of the preparations was conducted in a testing hospital laboratory after which one formulation was selected as being suitable for use. This was followed by writing a paper for open-access distribution to others needing to prepare lysis buffer elsewhere and the preparation of larger amounts of buffer to deliver to a hospital laboratory for ongoing distribution throughout Ireland.