Low LOD 1-Step RT-qPCR Mix

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

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12 12

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16 16

18 18 26

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22

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34

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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

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6

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