Supernatant was harvested after 7 days and purified using Ni-NTA agarose beads. Lateral flow immunoassays (LFIA) We tested seven point-of-care colloidal-gold-based LFIAs detecting IgG and IgM antibodies against SARS-CoV-2, full details of which are shown in Table 2. and 24 RT-PCR-confirmed SARS-CoV-2 positive samples were used as positive controls for sensitivity calculations. 95% CIs are shown for each calculation.(DOCX) ppat.1008817.s003.docx (37K) GUID:?E03AF901-33A3-4609-AB90-A69D8BEF8587 S2 Table: Head-to-head specificity calculations for all immunoassays. 50 pre-pandemic negative samples from the St Thomas emergency admissions cohort (STH Healthy, March 2019) were used to perform head-to-head specificity calculations for all immunoassay platforms. An extended panel of 105 samples was used for specificity calculations of the in-house ELISA for anti-S IgM and IgG. 95% CIs are shown for each calculation.(DOCX) ppat.1008817.s004.docx (34K) GUID:?830FFD8B-155C-48F7-A18C-4FF2B3171E5C S3 Table: Specificity of selected lateral flow immunoassays determined on an extended panel of pre-pandemic serum samples from March 2019. A select group of best-performing lateral flow immunoassays were taken forward for extended specificity calculations, using up to 200 pre-pandemic negative samples from the St Thomas emergency admissions cohort (STH Healthy, March 2019). 95% CIs are shown for each calculation.(DOCX) ppat.1008817.s005.docx (27K) GUID:?8DD00630-69AB-4C26-8990-DBD9A8580814 S4 Table: Sensitivity of immunoassays classified by days POS. Head-to-head sensitivity calculations were performed for all immunoassays PX-866 (Sonolisib) on a panel of 110 SARS-CoV-2-positive samples. Results for each test were further categorised according to whether the serum sample was from <10, 10, 14, or 20 days POS. 95% CIs are shown for each calculation.(DOCX) ppat.1008817.s006.docx (53K) GUID:?D014338E-89CB-4C83-B3B3-A29E60C79E26 S5 Table: Sensitivity of immunoassays classified by disease severity. Head-to-head sensitivity calculations were performed for all immunoassays on a panel of 110 SARS-CoV-2-positive samples and classified according to severity of disease, with 0 indicating mild illness (requiring no respiratory support) and 5 indicating critical (requiring ECMO) (see PX-866 (Sonolisib) Materials and methods for full classification). 95% CIs are shown for each calculation.(DOCX) ppat.1008817.s007.docx (47K) GUID:?71E835C2-492A-47AB-94D9-EDA2E6C81936 Data Availability StatementAll relevant data are within the manuscript and its Supporting Information files. Abstract There is a clear requirement for an accurate SARS-CoV-2 antibody test, both as a complement to existing diagnostic capabilities and for determining community seroprevalence. We therefore evaluated the performance of a variety of antibody testing technologies and their potential use as diagnostic tools. Highly specific in-house ELISAs were developed for the detection of anti-spike (S), -receptor binding domain (RBD) and -nucleocapsid (N) antibodies and used for the cross-comparison of ten commercial serological assaysa chemiluminescence-based platform, two ELISAs and seven colloidal gold lateral flow immunoassays (LFIAs)on an identical panel of 110 SARS-CoV-2-positive samples and 50 pre-pandemic negatives. There was a wide variation in the performance of the different platforms, with specificity ranging from 82% to 100%, and overall sensitivity from 60.9% to 87.3%. However, the head-to-head comparison of multiple sero-diagnostic assays on identical sample sets revealed that performance is highly dependent on the time of sampling, with sensitivities of over 95% seen in several tests when assessing samples from more than 20 days post onset of symptoms. Furthermore, these analyses identified clear outlying samples that were negative in all tests, but were later shown to be from individuals with mildest disease presentation. Rigorous comparison PX-866 (Sonolisib) of antibody testing platforms will inform the deployment of point-of-care technologies in healthcare settings and their use in the monitoring of SARS-CoV-2 infections. Author PX-866 (Sonolisib) summary PCR-based throat and nose swab tests for novel coronavirus (SARS-CoV-2) establish if someone is infected with the virus, while antibody tests can determine whether someone has had it in the past. However, for diagnosis Rabbit polyclonal to STOML2 later in disease, or in delayed-onset syndromes such as paediatric inflammatory multisystem syndrome (PIMS), antibody tests could form an important part of hospital diagnostic capabilities. They will also be essential for patient management strategies and community seroprevalence studies. We have conducted unbiased, head-to-head comparisons of ten commercial antibody test kits,.