Doherty Institute 2020 [A].

Study characteristics
Patient Sampling	Purpose: An updated report evaluating the diagnostic performance of three serological point‐of‐care tests and comparing these with two POC tests and one EIA test included in a previous report Multi‐group study to estimate sensitivity and specificity for diagnosis of active disease or identification of previous disease. Design: [1] Sensitivity group: patients with SARS‐CoV‐2 detected by RT‐PCR from upper and/or lower respiratory tract specimens. (n = 91 patients, 137 samples) [2] Specificity group: (n = 92 people, 92 samples) [2a] patients with infections with the potential for cross‐reactivity in serological assays, namely (i) patients with respiratory viral infections, including seasonal coronavirus infections and (ii) patients with other acute infections (e.g. dengue; CMV; EBV) (n = 36 patients, 36 samples) [2b] representative sample of the Victorian population collected in 2018 and 2019 (‘pre‐pandemic controls’) (n = 56 people, 56 samples) Recruitment: All serum samples were obtained from a tertiary hospital (Royal Melbourne Hospital, RMH) or the state reference laboratory for virology (Victorian Infectious Diseases Reference Laboratory, VIDRL). Prospective or retrospective: Retrospective Sample size: Patients: 183 (91) Samples: 229 (137) Further detail: Not stated
Patient characteristics and setting	Setting: Tertiary hospital and state reference laboratory Location: tertiary hospital (Royal Melbourne Hospital, RMH) or the state reference laboratory for virology (Victorian Infectious Diseases Reference Laboratory, VIDRL) Country: Australia Dates: Not stated Symptoms and severity: Not stated Demographics: Not stated Exposure history: Not stated Non‐Covid group 1: [2a] Other non‐COVID infections Source: Other diseases, dates not stated; pre‐pandemic controls 2018‐2019 Characteristics: Not stated Non‐Covid group 2: [2b] Pre‐pandemic controls Source: 2018‐2019 Characteristics: NR
Index tests	Test name: [A] Hangzhou Alltest IgG/IgM Rapid Test [B] Hangzhou unlabelled packaging (see comments) [C] Wondfo SARS‐CoV‐2 Antibody Test [D] Hightop SARS‐COV‐2 IgM/IgG Antibody Rapid Test [E] OnSite COVID‐19 IgG/IgM Rapid Test [F] VivaDiag COVID‐19 IgM/IgG Rapid Test [G] EUROIMMUN Anti‐SARS‐CoV‐2 ELISA (IgA) (IgG) Manufacturer: Not reported, but as per test names Antibody: [A] to [F] IgG, IgM, (NB assay [C] does not differentiate between antibody class, with only a single test line indicative of a positive test IgM/IgG) [G] IgA, IgG Antigen target: [A to F] The specific SARS‐CoV‐2 recombinant antigen(s) incorporated into the assay were not described in the manufacturers' information [G] Not stated Evaluation setting: [A to F] POC, used in laboratory; [G] Laboratory, used in laboratory Test method: [A to F] Lateral flow immunoassay (colloidal gold) (CGIA); [G] ELISA Timing of samples: 0 ‐> 30 days post‐symptom onset 0‐3 days pso: 23/137 (16.8%) samples 4‐8 days pso: 28/137 (20.4%) samples 9‐14 days pso: 21/137 (15.3%) samples 15‐20 days pso: 8/137 (5.8%) samples 21‐30 days pso: 27/137 (19.7%) samples > 30 days pso: 30/137 (21.9%) samples Samples used: Serum Test operator: [A to F] three laboratory research technicians, all of whom had undergone previous training in the use of lateral flow assays [G] Not reported Definition of test positivity: [A, B, D to F] Visible lines for IgG/IgM; [C] single test line indicative of a positive test (IgM/IgG); [G] Not reported [C] and [D] Testing was undertaken in duplicate, with a third test undertaken for discordant results.The majority result (i.e. 2/3) was taken as the final result, any faint line present at test termination was considered a positive result. Blinding reported: Yes. Threshold predefined: [A to F] Visible lines, interpreted as per manufacturer's instructions for use; [G] Not reported
Target condition and reference standard(s)	Reference standard: SARS‐CoV‐2 detected using the Coronavirus Typing assay (AusDiagnostics, Mascot, NSW) ‐ a two‐step, hemi‐nested multiplex tandem PCR In addition, all positive samples had SARS‐CoV‐2 detected at VIDRL where testing was first conducted using an in‐house assay for the SARS‐CoV‐2 RdRp gene. If positive, subsequent testing for the SARS‐CoV‐2 E gene was performed, using previously published primers. Samples used: Upper and/or lower respiratory tract specimens Timing of reference standard: Not stated Blinded to index test: Yes, prior Incorporated index test: No Definition of non‐COVID cases: [2a] Unclear for other diseases, [2b] NA for pre‐pandemic controls Samples used: [2a] Unclear for other diseases, [2b] NA for pre‐pandemic controls Timing of reference standard: [2a] Unclear for other diseases, [2b] NA for pre‐pandemic controls Blinded to index test: Yes, prior Incorporated index test: No
Flow and timing	Time interval between index and reference tests: Not stated All patients received same reference standard: No ‐ pre‐pandemic controls included Missing data: Not stated Uninterpretable results: Not stated Indeterminate results: One sample was excluded from testing in the Hightop SARS‐CoV‐2 IgM/IgG Antibody Rapid Test assay as results were discordant and insufficient test kits remained to test in triplicate. Unit of analysis: Samples
Comparative
Notes	Funding: Not stated Publication status: Report of post‐market validation (Report prepared for Office of Health Protection, Commonwealth Government of Australia, and the Therapeutics Goods Administration (TGA) of Australia) Source: Doherty Institute Author COI: Not stated
Methodological quality
Item	Authors' judgement	Risk of bias	Applicability concerns
DOMAIN 1: Patient Selection
Was a consecutive or random sample of patients enrolled?	Unclear
Was a case‐control design avoided?	No
Did the study avoid inappropriate exclusions?	Unclear
Did the study avoid inappropriate inclusions?	No
Could the selection of patients have introduced bias?		High risk
Are there concerns that the included patients and setting do not match the review question?			High
DOMAIN 2: Index Test (All tests)
DOMAIN 2: Index Test (Antibody tests)
Were the index test results interpreted without knowledge of the results of the reference standard?	Yes
If a threshold was used, was it pre‐specified?	Yes
Could the conduct or interpretation of the index test have introduced bias?		Low risk
Are there concerns that the index test, its conduct, or interpretation differ from the review question?			Unclear
DOMAIN 3: Reference Standard
Is the reference standards likely to correctly classify the target condition?	Unclear
Were the reference standard results interpreted without knowledge of the results of the index tests?	Yes
The reference standard does not incorporate the index test	Yes
Could the reference standard, its conduct, or its interpretation have introduced bias?		Unclear risk
Are there concerns that the target condition as defined by the reference standard does not match the question?			High
DOMAIN 4: Flow and Timing
Was there an appropriate interval between index test and reference standard?	Unclear
Did all patients receive the same reference standard?	No
Were all patients included in the analysis?	Unclear
Did all participants receive a reference standard?	Unclear
Were results presented per patient?	No
Could the patient flow have introduced bias?		High risk