Clinical Comparative Evaluation of the LabTurbo^TM AIO^® Reverse Transcription-Polymerase Chain Reaction and World Health Organization-Recommended Assays for the Detection of Emerging SARS-CoV-2 Variants of Concern

Figures & data

Table 1 Primer and Probe Sequences Used for Real-Time Reverse Transcription-Polymerase Chain Reaction (PCR) to Detect Severe Acute Respiratory Syndrome Coronavirus 2

	Target Gene	Primer Name	Sequence (5′→3′)	Reference
Single-plex PCR	E	E_Sarbeco_F	ACAGGTACGTTAATAGTTAATAGCGT	[12]
		E_Sarbeco_R	ATATTGCAGCAGTACGCACACA
		E_Sarbeco_P1	FAM-ACACTAGCCATCCTTACTGCGCTTCG-BBQ
	RdRp	RdRp_SARSr-F2	GTGARATGGTCATGTGTGGCGG
		RdRp_SARSr-R2	CAAATGTTAAAAACACTATTAGCATA
		RdRp_SARSr-P2	FAM-CAGGTGGAACCTCATCAGGAGATGC-BBQ
Multiplex PCR	E	E_Sarbeco_F	ACAGGTACGTTAATAGTTAATAGCGT	[12]
		E_Sarbeco_R	ATATTGCAGCAGTACGCACACA
		E_Sarbeco_P1	/5HEX/ACACTAGCC/ZEN/ATCCTTACTGCGCTTCG/3IABkFQ/
	N1	2019-nCoV_N1-F	GACCCCAAAATCAGCGAAAT	[13]
		2019-nCoV_N1-R	TCTGGTTACTGCCAGTTGAATCTG
		2019-nCoV_N1-P	/56-FAM/ACCCCGCAT/ZEN/TACGTTTGGTGGACC/3IABkFQ/
Single-plex PCR	RNase P	RP-F	AGATTTGGACCTGCGAGCG	[13]
Multiplex PCR		RP-R	GAGCGGCTGTCTCCACAAGT
		RP-P	/5Cy5/TTCTGACCT/TAO/GAAGGCTCTGCGCG/3IAbRQSp/

Table 2 Limit of Detection of LabTurbo^TM AIO COVID-19 RNA Testing Kit for N1 and E Using the Rotor-Gene-Q Real-Time PCR Instrument

	Target Gene
Viral load (copies/reaction)	N1	E
4.7	N.D.^a	N.D.
	N.D.	N.D.
	N.D.	N.D.
	N.D.	N.D.
	N.D.	N.D.
9.4	35.6	34.9
	33.5	33.3
	33.8	33.8
	34.1	34.6
	34.0	33.5
	33.9	34.3
	32.8	33.5
	35.5	34.8
	34.0	33.9
	35.5	34.9
	34.5	34.9
	35.7	34.0
	32.8	33.0
	33.9	34.8
	34.4	33.8
	34.6	36.1
	32.9	34.8
	34.5	33.8
	34.0	35.1
	35.6	34.9
18.75	32.9	32.8
	32.6	33.7
	33.5	35.8
	32.7	33.1
	34.0	34.4
	32.3	33.3
	32.9	33.4
	33.5	34.3
	33.2	33.8
	33.0	33.3
	32.9	32.7
	32.9	33.1
	32.5	32.9
	34.4	33.5
	32.8	32.6
	33.9	33.8
	32.4	32.3
	33.9	33.3
	32.7	33.8
	32.9	34.3
37.5	31.6	31.3
	31.3	31.0
75	30.2	30.6
	30.2	30.3
150	29.0	29.1
	29.0	29.0
300	28.2	28.3
	28.1	28.1

Abbreviation: ^aN.D., not detected.

Table 3 Cross-Reaction of the LabTurbo^TM AIO COVID-19 RNA Testing Assay with Known Respiratory Viruses in Clinical Samples or Cell Supernatants

Viral Pathogens	LabTurbo^TM AIO COVID-19 RNA Testing Assay
Influenza A (H1)	Not detected
Influenza A (H3)	Not detected
Influenza A (H1N1)	Not detected
Influenza B	Not detected
Respiratory syncytial virus	Not detected
Rhinovirus/ Enterovirus	Not detected
Parainfluenza 1 virus	Not detected
Parainfluenza 2 virus	Not detected
Parainfluenza 3 virus	Not detected
Adenovirus	Not detected
Coronavirus NL96	Not detected

Figure 1 Correlation between results obtained using LabTurbo^TM AIO COVID-19 RNA testing assay and WHO-recommended assay with the Rotor-Gene-Q real-time PCR instrument. (A) SARS-CoV-2 N1 versus RdRp screening results. (B) SARS-CoV-2 E screening results; correlation between results obtained using LabTurbo^TM AIO COVID-19 RNA testing assay and WHO-recommended assay with the Roche LightCycler96 instrument. (C) Shows SARS-CoV-2 N1 versus RdRp gene results. (D) Shows SARS-CoV-2 E gene results.

Corman VM, Landt O, Kaiser M, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020;25. doi:10.2807/1560-7917.ES.2020.25.3.2000045

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Centers for disease control and prevention. 2019-novel coronavirus (2019-nCoV) real-time rRT-PCR panel primers and probes; 2020. Available from:https://www.cdc.gov/coronavirus/2019-ncov/downloads/rt-pcr-panel-primer-probes.pdf. Accessed May 29, 2020.

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