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FIND evaluations of SARS-CoV-2 molecular tests

In February 2020, FIND launched an expression of interest (EOI) for test developers of in vitro diagnostics (IVDs) that detect SARS-CoV-2 nucleic acid (molecular tests) to participate in independent evaluation studies. Over 200 submissions were received and applications were reviewed according to the following scoring criteria (Table 1).

Table 1: Scoring criteria
Max score
Limit of detection3
Regulatory status2
Type of organization1
Quality management system1
Other products available in low- and middle-income countries3
TOTAL10

As of August 2020, FIND is no longer accepting applications to evaluate molecular tests.

FIND conducted independent evaluations at the University Hospitals of Geneva (HUG) to verify the limit of detection (LOD) – as reported by the manufacturers – and the clinical performance of 22 manual molecular test kits in comparison to an in-house PCR protocol that was optimized based on the Tib Molbiol assay. The LOD analysis was performed using cultured viral stocks from a clinical isolate from Switzerland that was quantified using an E gene standard. The clinical performance analysis was conducted on extracted samples from individuals suspected to have COVID-19, 50 of which were reference PCR positive and 100 of which were reference PCR negative. Data are summarized in Table 2.

Additionally, a limited clinical performance evaluation of the Cepheid Xpert Xpress SARS-CoV-2 assay was also performed at HUG. A second collaborating site, the Translational Health Science and Technology Institute (THSTI) conducted a similar limited clinical performance evaluation of the Molbio TrueNat SARS-CoV-2 assay. Both studies were performed using frozen, stored respiratory samples from COVID-19 suspects. Results on the performance of these automated near-POC assays are shown in Table 3.

COVID-19 molecular assay evaluation protocol

Read the protocol
Table 2: Results for 22 manual (open) molecular tests evaluated at HUG
CompanyProduct nameProduct numberGene target Verified LOD (copies / reaction) Avg Ct (lowest dilution 10/10) Clinical sensitivity (50 positives)Clinical specificity*
(100 negatives)		Lot No.PCR platform**Supplier recommended Ct cut-off
1.altona DiagnosticsRealStar®
SARS-CoV-2
RT-PCR Kit 1.0		821003/821005E1–1035.4592%
(95%CI:
81, 97)		100%
(95%CI:
96, 100)		023567BioRad CFX96 deep wellNone;
any signal can be considered positive
S1–1035.9992%
(95%CI:
81, 97)		100%
(95%CI:
96, 100)
2.Atila BioSystems Inc.Atila iAMP
COVID-19 Detection (isothermal detection)		iAMP-COVID-100-RUOORF1ab50–100N/A100%
(95%CI:
93, 100)		99%*
(95%CI:
95, 100)		COVID20200320BioRad CFX96 deep wellAny signal is considered positive (isothermal)
N1–10N/A100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
3.Beijing Wantai Biological Pharmacy Enterprise Co. LtdWantai
SARS-CoV-2
RT-PCR Kit		WS-1248ORF1ab1–1036.20100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		nCoVP20200305BioRad CFX96 deep well≤40
N1–1037.12100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
4.BGI Health (HK) Co. LtdReal-time Fluorescent
RT-PCR kit for detection 2019-nCOV (CE-IVD)		MFG030010ORF11–1032.43100%
(95%CI:
93, 100)		99%*
(95%CI:
95, 100)		6220200305Roche LightCycler 480≤38
5.bioMérieux SAARGENE®
SARS-COV-2
R-GENE®[b]		423720
(CE-IVD)
423717 (RUO)		N10–5036.44100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		1007989610
1007947520		BioRad CFX96 deep wellAny signal considered as positive
RdRP10–5032.4496%[a]
(95%CI:
87, 99)		100%
(95%CI:
96, 100)
6.Bioneer Corporation

AccuPower®
SARS-CoV-2 Real-Time

RT-PCR Kit		SCV-2122E10–5035.85100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		200931EBioRad CFX96 deep well<38
RdRP10–5036.18100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
7.Boditech Med. Inc.ExAmplar COVID-19

real-time
PCR kit (L)		UFPK-4E10–5034.9100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		WLQCB02LBioRad CFX96 deep well≤42
RdRP50–10033.4690%
(95%CI:
79, 96)		100%
(95%CI:
96, 100)
8.CerTest Biotec S.L.VIASURE SARS-CoV-2 Real Time PCR Detection KitVS-NC0112L
VS-NC0212L		ORF1ab10–5035.1698%
(95%CI:
90, 100)		100%
(95%CI:
96, 100)		NCO212L-023BioRad CFX96 deep well<40
N1–1035.46100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
9.DAAN Gene Co. Ltd of Sun Yat-Sen UniversityDetection Kit for 2019 Novel Coronavirus (2019-nCoV) RNA (PCR-Fluorescence Probing)DA0930-DA0932ORF11–1038.76100%
(95%CI:
93, 100)		96%*
(95%CI:
90, 98)		2020007Roche LightCycler 480≤40
N1–1036.97100%
(95%CI:
93, 100)		98%*
(95%CI:
93, 99)
10.EUROIMMUN AGEURORealTime SARS-CoV-2[c]MP 2606-0425ORF1ab/N1–1037.88100%
(95%CI:
93, 100)		98%*
(95%CI:
93, 99)		I200320ALLight Cycler 480 IIAny signal considered positive
11. GeneFirst LtdThe Novel Coronavirus (2019-nCoV) Nucleic Acid Test KitMPA-COVID19ORF11–1035.45100%
(95%CI:
93, 100)		99%*
(95%CI:
95, 100)		00072BioRad CFX96 deep well≤37.0 positive;

37-40 indeterminate; >40 negative
N1–1036.7298%
(95%CI:
90, 100)		100%
(95%CI:
96, 100)
12.KH Medical Co. LtdRADI
COVID-19 Detection Kit		RV008S1–1037.94100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		V008.200202BioRad CFX96 deep well≤40
RdRP10–5036.74100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
13.PerkinElmer Inc.PerkinElmer® SARS-CoV-2 Real-time
RT-PCR Assay[c,d]		SY580N1–1039,43100%
(95%CI:
93, 100)		99%*
(95%CI:
95, 100)		8220200303BioRad CFX96 deep well≤42
ORF11–1038,99100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
14.Primerdesign Ltd

Coronavirus COVID-19 genesig®
Real-Time PCR assay[c]		Z-Path-COVID-19-CERdRP1–1036.7100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		JN-02780-0009LightCycler 480Any signal regarded as positive
15.QuantumDxQuantuMDx SARS-CoV-2 RT-PCR Detection AssayQ22003Orf1, N, S1–1036.8100%
(95% CI:
92, 100)		100%
(95% CI:
96, 100		P01100BioRad CFX96 deep well≤40
16.R-Biopharm AGRIDA®GENE SARS-CoV-2 RUOPG6815RUOE1–1037.99100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		21120NBioRad CFX96 deep wellNone;
any signal can be considered positive
17.Sansure Biotech Inc.Novel Coronavirus (2019-nCoV) Nucleic Acid Diagnostic Kit (PCR-Fluorescence Probing)[e]S3102EORF110–5035.16100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		2020007ZCThermofisher Quantstudio 5≤40
N10–5034.96100%
(95%CI:
93, 100)		95%*
(95%CI:
89–98)
18.SD Biosensor Inc.STANDARD M nCoV
Real-Time Detection Kit		M-NCOV-01E1–1037.43100%
(95%CI:
93, 100)		97%*
(95%CI:
92, 99)		MNC00120005Roche LightCycler 480≤41
ORF11–1036.99100%
(95%CI:
93, 100)		99%*
(95%CI:
95, 100)
19.Seegene Inc.Allplex™
2019-nCoV Assay		RP10244Y
RP10243X		E1–1033.3100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		RP4520C24BioRad CFX96≤40
N1–1036.74100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
RdRP1–1034.73100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
20.Shanghai Kehua Bio-Engineering Co. LtdKHB Diagnostic kit for
SARS-CoV-2 Nucleic Acid (Real-time PCR)		KH-G-M-574-48ORF11–1030.39100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		20037410BioRad CFX96 deep wellMore than two targets detected and curve is of S shape
N1–1032.95100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
E1–1031.72100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
21.ThermoFisher ScientificTaqPath™ COVID-19
CE-IVD
RT-PCR Kit[f]		A48067ORF1ab;
S
N		1–10NA100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)		2225262Quantstudio 5Not Applicable (Automated software interpretation)
22.Vela DiagnosticsViroKey™ SARS-CoV-2 RT-PCR Test[c]300682RdRP10–5030.9594%
(95%CI:
84, 98)		100%
(95%CI:
96, 100)		1000000597BioRad CFX96 deep well≤40
ORF11–1035.57100%
(95%CI:
93, 100)		100%
(95%CI:
96, 100)
Tib Molbiol/Roche DiagnosticsModularDx Kit SARS-CoV (COVID19) E-gene (Tib Molbiol) + LightCycler Multiplex RNA Virus Master (Roche)53-0776-96
6754155001		E1–1033.34100%
(95%CI:
93, 100)		100%
96, 100)		48202019
48274100		Roche LightCycler 480Define the
cut-off 2–4 cycles higher than observed Cp value for

10 copies
* Clinical specificity: Further investigation is needed to determine if apparent false positives are truly false positives or whether they are due to a false negative reference standard result
** PCR platform: All products were evaluated on a PCR platform recommended by the supplier, listed in this table. Each test can be performed on other PCR systems, detailed in the product’s instructions for use.
[a] The two false negative samples tested positive with the second PCR (PCR 2) that targets E gene of SARS, SARS-COV-2 and/or SARS-like coronaviruses.
[b] Samples for both analytical and clinical analyses were from already-extracted specimen, therefore the methods varied from those recommended by the supplier as the internal control was not included.
[c] Samples for both analytical and clinical analyses were from already-extracted specimen, therefore the methods varied from those recommended by the supplier as the internal control was added to the master mix.
[d] Evaluation procedure varied from recommended protocol. In order to achieve the recommended sample input volume, a 2.5 fold dilution of the samples was used.
[e] Sansure claims a lower LOD of 6.4 cp/rxn, which has been independently verified.
[f] Evaluation procedure varied from recommended protocol as source material was already-extracted RNA; extracted MS2 control was added directly to the master mix.
Table 3: Results for evaluation of two near-POC automated tests
CompanyProduct nameProduct numberGene target Clinical sensitivity (50 positives) Clinical specificity*
(100 negatives)		Comparator test
Cepheid Inc.Xpert® Xpress
SARS-CoV-2		XPRSARS-COV2-10N2100%
(95%CI:
92,100)		99% *
(95%CI:
95, 100)		Roche Cobas ® SARS-CoV-2
N = 44 positive
N = 100 negative
E97.7%
(95% CI:
88, 100)		100%
(95%CI:
96, 100)
Molbio Diagnostics Pvt LtdTrueNat
SARS-CoV-2[1]		601410020E+RdRP[2]98%
(95% CI:
90.98)		96% *
(95% CI:90,98)		altona Diagnostics (n=86) /LabGun™ (n=64) and/or Seegene, Inc. (n=12)
N = 51 positive
N = 111 negative
601420050
* Clinical specificity: Further investigation is needed to determine if apparent false positives are truly false positives or whether they are due to a false negative reference standard result
[1] Note: evaluation performed at THSTI

[2] RdRP is only used as a reflex test; the results are for combined E+RdRP positives

More information

For questions relating to the evaluation of molecular tests, please contact our Emerging Threats team.