Low LOD 1-Step RT-qPCR Mix

Low LOD 1-Step RT-qPCR Mix

Specialized for automated, multiplex assays detecting blood-borne viruses for blood bank and organ donor testing

Suitable Application

Blood Banking

Transmission of viral infections through organ transplants and the transfusion of blood products has been known for decades but was considered to be an unavoidable consequence of a life- saving treatment. Molecular testing has helped reduce the number of viral transfusion-transmitted infections due to its superior sensitivity and specificity over other testing methods. However, unique to blood banking and organ donor testing is the challenge of detecting viruses during the window period before seroconversion. PCR inhibitors traditionally found in blood samples have posed a significant challenge in developing molecular assays that can detect low level viremia in early stage infections. Meridian’s new Low LOD 1-Step RT-qPCR Mix has been specifically developed for highly reproducible first-strand cDNA synthesis and qPCR in a single tube. It is formulated with an antibody-mediated hot-start DNA polymerase and uses the latest advances in buffer chemistry and enhancers. Containing dUTP, it is ideal for automated multiplex testing of viruses such as HCV, HBV, HIV-1, HIV-2, HAV and Parvovirus B19. Low LOD 1-Step RT-qPCR Mix has already been successfully adopted for a CE-IVD Marked NAT Blood Banking assay.

Low LOD 1-Step RT-qPCR Mix • Suitable for detecting purified RNA and DNA viruses at very low titers • Novel buffer system optimized for multiplex assays • Perform first-stand DNA synthesis and qPCR in a single tube

PRODUCT

CAT NO.

VOLUME REACTIONS

10 mL

1,000 Rxn

Low LOD 1-Step RT-qPCR Mix

MDX025

100 mL

10,000 Rxn

www.MeridianLifeScience.com

Product Highlights

Figure 1: Sensitive multiplex qPCR for simultaneous detection of DNA and RNA viruses 22 replicates of each extract dilution were tested in multiplex qPCR reactions using the Low LOD 1-Step RT-qPCR Mix and the limit of detection hit rate was determined as a percentage of positive amplifications. The multiplex reactions were HAV, HBV, HCV and an extraction control and HIV-1, HIV-2, Parvovirus B19 and an extraction control. Results illustrate that Low LOD 1-Step RT-qPCR Mix can be used in multiplex reactions with both RNA and DNA viral templates, making it suitable for monitoring viral load for different types of virus. Sensitive and reproducible detection of low-level viremia for both RNA viruses (HCV, HIV-1, HIV-2 and HAV) and DNA viruses (HBV and Parvovirus B19).

HCV (RNA Virus)

HIV-1 (RNA Virus)

HAV (RNA Virus)

100

100

100

75

75

75

50

50

50

25

25

25

0

0

0

150 IU/mL 75 IU/mL 37.5 IU/mL 18.75 IU/mL 9.375 IU/mL 4.68 IU/mL

500 IU/mL 250 IU/mL 125 IU/mL 62.5 IU/mL 31.3 IU/mL 15.6 IU/mL

30 IU/mL

15 IU/mL

7.5 IU/mL 3.75 IU/mL 1.9 IU/mL 0.9 IU/mL

HIV-2 (RNA Virus)

B19 (DNA Virus)

HBV (DNA Virus)

100

100

100

75

75

75

50

50

50

25

25

25

0

0

0

30 IU/mL

15 IU/mL

7.5 IU/mL 3.75 IU/mL 1.9 IU/mL 0.9 IU/mL

500 IU/mL 250 IU/mL 125 IU/mL 62.5 IU/mL 31.3 IU/mL 15.6 IU/mL

60 IU/mL

30 IU/mL

15 IU/mL

7.5 IU/mL 3.75 IU/mL 1.9 IU/mL

Figure 2: Low limit of detection (LOD) for both RNA and DNA virus 24 replicates of purified blood viral extracts were tested in singleplex qPCR reactions using the LowLOD 1-Step RT-qPCR Mix at the indicated IU/mL. The singleplex reactions were HAV, HBV, HCV, HIV-1, HIV-2, Parvovirus B19. The amplification curve results illustrate that Low LOD 1-Step RT-qPCR Mix can be used with both RNA and DNA viruses as low as 5 IU/mL.

Amplification Curves plification Curves HCV (30 IU/mL) Amplification Curves

Amplification Curves HBV (15 IU/mL)

Amplification Curves Amplification Curves HIV-1 (60 IU/mL)

Amplification Curves

23.688 23.079 23.079 23.6 8

63.075

23.688

21.688 21.079 21.079 21.6 8

21.688

57.075

19.688 19.079 19.079 19.6 8

19.688

51.075

17.688 17.079 17.079 17.6 8

17.688

45.075

15.688 15.079 15.079 15.6 8

15.688

39.075

13.688 13.079 13.079 13.6 8

13.688

33.075

11.688 11.079 11.079 1.6 8

11.688

27.075

9.688 9.079 9.079 9.6 8

9.688

21.075

7.688 7.079 7.079 7.6 8

7.688

15.075

5.688 5.079 5.079 3.688 3.079 3.079 3.6 8 1.688 1.079 1.079 1.6 8 5.6 8

5.688

3.688

9.075

1.688

3.075

2 4 6 8 10 2 4 6 8 10 12

12 12

14 14 20 14

16 16

18 18 26

20 20

22 22

24 24 24

26 26

28 28

30 30

32 32 32

34 34 34

36 36 36

38 38 38

40 40 40

42 42 42

44 44 4

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

38

40

42

44

10 4 16

18

22

24

28

22 30 3

34

36

38

40

42

44

2

4

6

8

12

16

18

20

26

28

30

2

4

6

8

10

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

38

40

42

44

Cycles

Cycles

Cycles

Cycles

Cycles

Cycles

Cycles

Amplification Curves Amplification Curves HIV-2 (5 IU/mL)

Amplification Curves HAV (5 IU/mL) Amplification Curves

B19 (60 IU/mL)

Amplification Curves

Amplification Curves

6.628

40.432

43.077

36.932

6.028

39.077

33.432

5.428

35.077

29.932

4.828

31.077

26.432

4.228

27.077

22.932

3.628

23.077

19.432

3.028

19.077

15.932

2.428

15.077

12.432

1.828

11.077

8.932

1.228

7.077

5.432

0.628

3.077

1.932

0.028

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

38

40

42

44

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

38

40

42

44

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

38

40

42

44

Cycles

Cycles

Cycles

Cycles

Cycles

Cycles

Ordering information: USA E: info@meridianlifescience.com Toll free: +1 800 327 6299 UK E: info.uk@meridianlifescience.com Tel: +44 (0)20 8830 5300

Germany E: info.de@meridianlifescience.com Tel: +49 (0)3371 60222 00 France E: info.fr@meridianlifescience.com Tel: +33 (0)1 42 56 04 40

美国迈迪安生命科学公司 电子邮件: vivian.li@meridianlifescience.com 电话: +86-159-1103-0750

Connect with us:

Australia E: info.au@meridianlifescience.com Tel: +61 (0)2 9209 4180

www.MeridianLifeScience.com

ISO 13485 Certified

10/19

Page 1 Page 2

meridianlifescience.com

Powered by