Xiamen Zeesan Biotech Co.，Ltd.

Add: Building 1, 3701 Xiang'an North Road, Torch Hi-Tech Industrial

Development Zone (Xiang'an), Xiamen City, Fujian Province, P.R.
China
http://www.zsandx.com P.C:361101

Xiamen Zeesan Biotech Co., Ltd.

 Xiamen Zeesan Biotech Co.，Ltd.
Add: Building 1, 3701 Xiang'an North Road, Torch Hi-Tech Industrial
Development Zone (Xiang'an), Xiamen City, Fujian Province, P.R.
China
http://www.zsandx.com P.C:361101

With the evolution of SARS-CoV-2, the genes of SARS-CoV-2 are mutating all the time. Thus,
the identification accuracy is being challenged.

Zeesan SARS-CoV-2 Test Kit targets at the specific conserved sequences of ORF1ab and N
Gene. According to the sequence alignment, the mutants of ORF1ab and N Gene in the major
variants (B.1.1.7/B.1.351/P.1/B.1.617.1/B.1.617.2) are not in the area covered by primers or
probes (Figure 1). Therefore, the accuracy in identifying the variants by the kits is not affected.

Figure 1: The Target Area of Zeesan SARS-CoV-2 Test Kit and the Mutation Sites

Note: (1) means the target area of Zeesan SARS-CoV-2 Test Kit.
(2) Although C28977T mutation is in the target area of N Gene, the mutation is not in the area
covered by primers or probes. The accuracy in identifying the variants by the kits is not
affected.

Wild-Type SARS-CoV-2 Positive Sample

SARS-CoV-2 Variant B.1.1.7 Positive Sample

Figure 2: CT Values of ORF1ab (Sample Concentration: 200copies/ml)

 Xiamen Zeesan Biotech Co.，Ltd.
Add: Building 1, 3701 Xiang'an North Road, Torch Hi-Tech Industrial
Development Zone (Xiang'an), Xiamen City, Fujian Province, P.R.
China
http://www.zsandx.com P.C:361101

Wild-Type SARS-CoV-2 Positive Sample

SARS-CoV-2 Variant B.1.1.7 Positive Sample

Figure 3: CT Values of N Gene (Sample Concentration: 200copies/ml)

Detailed Description:

1. B.1.1.7(20I/S:501Y.V1)

VARIANT GENE NUCLEOTIDE AMINO ACID

B.1.1.7 ORF1ab C3267T T1001I

C5388A A1708D
T6954C I2230T
11288-11296deletion SGF3675-3677deletion
C14408T P314L
N 28280GAT->CTA D3L
G28881A R203K
G28882A R203K
G28883C G204R
C28977T S235F
Table 1: The major mutations of SARS-CoV-2 Variant B.1.1.7 on ORF1ab and N Gene

Conclusion: Zeesan SARS-CoV-2 Test Kit targets at the specific conserved sequences of ORF1ab
and N Gene. According to the comparative analysis of biological information, SARS-CoV-2 Variant
B.1.1.7 has no mutation in the amplicon area of ORF1ab Gene. The only mutation(C28977T) in the
amplicon area of N Gene is not in the area covered by primers or probes, so the accuracy in
identifying the variants by the kits is not affected. In summary, the testing result of Zeesan
SARS-CoV-2 Test Kit remains reliable.
 Xiamen Zeesan Biotech Co.，Ltd.
Add: Building 1, 3701 Xiang'an North Road, Torch Hi-Tech Industrial
Development Zone (Xiang'an), Xiamen City, Fujian Province, P.R.
China
http://www.zsandx.com P.C:361101

2. B.1.351(20H/S:501Y.V2)

VARIANT GENE NUCLEOTIDE AMINO ACID

B.1.351 ORF1ab C1059T T265I

A2692T -
C3037T -
G5230T K1655N
A10323G K3353R
11288-11296deletion SGF3675-3677deletion
C14408T P314L
N C28887T T205I
Table 2: The major mutations of SARS-CoV-2 Variant B.1.351 on ORF1ab and N Gene

Conclusion: Zeesan SARS-CoV-2 Test Kit targets at the specific conserved sequences of ORF1ab
and N Gene. According to the comparative analysis of biological information, SARS-CoV-2 Variant
B.1.1.7 has no mutation in the amplicon area of ORF1ab Gene. The only mutation(C28977T) in the
amplicon area of N Gene is not in the area covered by primers or probes, so the accuracy in
identifying the variants by the kits is not affected. In summary, the testing result of Zeesan
SARS-CoV-2 Test Kit remains reliable.

3. P.1(20J/S:501Y.V3)

VARIANT GENE NUCLEOTIDE AMINO ACID

P.1 ORF1ab C3037T -

C3828T S1188L
A5648C K1795Q
11288-11296deletion SGF3675-3677deletion
C14408T P314L
G17259T E1264D
N C28512G P80R
G28881A R203K
G28882A R203K
G28883C G204R
Table 3: The major mutations of SARS-CoV-2 Variant P.1 on ORF1ab and N Gene

Conclusion: Zeesan SARS-CoV-2 Test Kit targets at the specific conserved sequences of ORF1ab
and N Gene. According to the comparative analysis of biological information, SARS-CoV-2 Variant
B.1.1.7 has no mutation in the amplicon area of ORF1ab Gene. The only mutation(C28977T) in the
 Xiamen Zeesan Biotech Co.，Ltd.
Add: Building 1, 3701 Xiang'an North Road, Torch Hi-Tech Industrial
Development Zone (Xiang'an), Xiamen City, Fujian Province, P.R.
China
http://www.zsandx.com P.C:361101

amplicon area of N Gene is not in the area covered by primers or probes, so the accuracy in
identifying the variants by the kits is not affected. In summary, the testing result of Zeesan
SARS-CoV-2 Test Kit remains reliable.

4. B.1.617.1(21A/S:154K)

VARIANT GENE NUCLEOTIDE AMINO ACID

B.1.617.1 ORF1ab C4965T T1567I

A11201G T3646A
C14408T P314L
G17523T M1352I
A20396G K2310R
N G28881T R203M
G29402T D377Y
Table 4: The major mutations of SARS-CoV-2 Variant B.1.617.1 on ORF1ab and N Gene

Conclusion: Zeesan SARS-CoV-2 Test Kit targets at the specific conserved sequences of ORF1ab
and N Gene. According to the comparative analysis of biological information, SARS-CoV-2 Variant
B.1.1.7 has no mutation in the amplicon area of ORF1ab Gene. The only mutation(C28977T) in the
amplicon area of N Gene is not in the area covered by primers or probes, so the accuracy in
identifying the variants by the kits is not affected. In summary, the testing result of Zeesan
SARS-CoV-2 Test Kit remains reliable.

5. B.1.617.2(21A/S:478K)

VARIANT GENE NUCLEOTIDE AMINO ACID

B.1.617.2 ORF1ab C14408T P314L

C16466T P1000L
N A28461G D63G
G28881T R203M
G29402T D377Y
Table 5: The major mutations of SARS-CoV-2 Variant B.1.617.2 on ORF1ab and N Gene

Conclusion: Zeesan SARS-CoV-2 Test Kit targets at the specific conserved sequences of ORF1ab
and N Gene. According to the comparative analysis of biological information, SARS-CoV-2 Variant
B.1.1.7 has no mutation in the amplicon area of ORF1ab Gene. The only mutation(C28977T) in the
amplicon area of N Gene is not in the area covered by primers or probes, so the accuracy in
identifying the variants by the kits is not affected. In summary, the testing result of Zeesan
SARS-CoV-2 Test Kit remains reliable.