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Aptima HCV RNA qualitative assay

The Aptima HCV RNA qualitative assay is FDA approved for the diagnosis of active hepatitis C virus (HCV) infection:

  • Amongst highest sensitivity of detection compared to other FDA approved qualitative HCV RNA assays.
  • Identify active HCV infections in individuals with antibody evidence and link to care using highly effective antiviral regimens to eradicate virus.
About

The Aptima HCV RNA Qualitative Assay is an in vitro nucleic acid amplification assay offering proven performance for the detection of HCV RNA in fresh or frozen human plasma (EDTA, sodium citrate and ACD) and serum:

  • Specific and sensitive detection across HCV genotypes 1 to 6 (and major subtypes).
  • Fully validated, transcription-mediated amplification (TMA) system with simple and controlled workflow without thermocycling.
  • The Aptima HCV RNA Qualitative Assay supports CDC testing algorithms for HCV diagnosis with the use of an FDA-approved nucleic acid amplification test (NAAT) intended for detection of HCV RNA in serum or plasma from blood of at-risk patients who test reactive for HCV antibody.1

Why is testing for current or active HCV infection important?

New treatments cure HCV infection but 75% of people with HCV don’t know they are infected due to mild course of disease. Without identifying active HCV infection, at least 70% of individuals are at risk for chronic infection, with potential for progression to cirrhosis, liver cancer and need for liver transplantation.2,3,4

Who should get tested for HCV according to CDC?1

Anyone who has injected drugs, or has certain pre-existing medical conditions (chronic liver disease or HIV), born between 1945 and 1965, health workers parentally exposed to blood, hemodialysis patients, and individuals born to HCV-infected mothers.

Features

The FDA-approved Aptima HCV RNA qualitative assay delivers proven performance:

  • Qualified in fresh or frozen human plasma (EDTA, sodium citrate and ACD) and serum.
  • Detects through genotypes 1 to 6 (and major subtypes).
  • Highly sensitive down to 9.6 IU/mL overall across genotypes.5

Our fully validated transcription-mediated amplification (TMA) Aptima HCV RNA qualitative assay offers:

  • Enhanced target detection.
  • Reduced carry-over contamination with more labile RNA amplification versus DNA.
  • Simple and controlled workflow with no thermocycling.
Performance Data

Table 1. Limit of Detection of HCV WHO International Standard

Concentration
(IU/ML)

 

Reactive

 

Non-Reactive

 

Invalid

 

Total

Percent Valids
Detected

 

95% CI*

0.1

1

59

0

60

1.7

0.3–8.9

1

23

36

1

60

39.0

27.6–51.7

2.5

178

61

1

240

74.5

68.7–79.7

5

220

19

1

240

92.1

87.9–94.9

7.5

236

2

2

240

99.2

97.0–99.8

10

60

0

0

60

100

94.0–100

18.5

59

0

1

60

100

94.0–100

50

60

0

0

60

100

94.0–100

The Limit of Detection (LOD) of the Aptima HCV RNA Qualitative Assay was determined by testing serial dilutions of the WHO International Standard for HCV genotype 1 RNA (NIBSC code 96/790).6 Each panel member was tested in replicates ranging from 60 to 240. Serial dilutions of the WHO International Standard for HCV genotype 1 RNA were detected 95% of the time as low as 7.5 IU/mL. Linear regression analysis determined 5.3IU/mL ≥(95% probability) as the limit of detection for the Aptima HCV RNA qualitative assay.

 

Table 2. HCV Genotype Detection (Transcripts)

HCV Genotype

Concentration
(IU/ML)
(copies/mL)

Reactive

Non-Reactive

Invalid

Total

Percent Valids
Detected

95% CI*

1

9.6 (50)

716

3

1

720

99.6

98.8–99.9

2a

9.6 (50)

357

3

0

360

99.2

97.6–99.7

2b

14.4 (75)

350

10

0

360

97.2

95.0–98.5

3a

9.5 (50)

356

4

0

360

98.9

97.2–99.6

4a

9.6 (50)

357

1

2

360

99.7

98.4–99.9

5a

9.6 (50)

356

4

0

360

98.9

97.2–99.6

6a

9.6 (50)

685

35

0

720

95.1

93.3–96.5

* Confidence Interval

Note: Invalid results were not included in the data calculations for Percent Valids Detected.

Transcripts of HCV genotypes 1, 2a, 2b, 3a, 4a, 5a, and 6a made from the 5’-untranslated region of the HCV genome were tested using the Aptima HCV RNA qualitative assay. All transcripts were quantitated using phosphate analysis and confirmed using hyperchromicity and OD260. The copies/mL were converted to IU/mL using an in-house conversion factor 5.2 copies/mL = 1IU/mL. Dilutions of each transcript were tested at 9.6 IU/mL (50 copies/mL) for genotypes 1, 2a, 3a, 4a, 5a, and 6a, and at 14.4 IU/mL (75 copies/mL) for HCV2b. Each transcript was tested in replicates ranging from 360 to 720.

 

Table 3. HCV Genotype Detection (Clinical Specimens)

HCV Genotype

Concentration (IU/ML) (copies/mL)

Total

Reactive

Non-Reactive

Invalid

Percent Valids
Detected

95% CI*

1

4.8 (25)

90

87

3

0

96.7

90.7–98.9

1

9.6 (50)

495

490

4

1

99.2

97.9–99.7

1

14.4 (75)

60

59

1

0

98.3

91.1–99.7

1

19.2 (100)

90

90

0

0

100

95.9–100

1

96.2 (500)

60

60

0

0

100

94.0–100

1

9,615.4 (50,000)

30

30

0

0

100

88.7–100

2b

9.6 (50)

210

200

10

0

95.2

91.5–97.4

2b

14.4 (75)

176

173

3

0

98.3

95.1–99.4

2b

19.2 (100)

120

120

0

0

100

96.9–100

2b

28.8 (150)

30

29

0

1

100

88.3–100

2b

38.5 (200)

90

90

0

0

100

95.9–100

2b

57.7 (300)

10

10

0

0

100

72.3–100

3a

9.6 (50)

120

120

0

0

100

96.7–100

3a

19.2 (100)

60

60

0

0

100

94.0–100

3a

38.5 (200)

40

40

0

0

100

91.2–100

4/4a

9.6 (50)

120

114

6

0

95.0

89.5–97.7

4/4a

14.4 (75)

30

30

0

0

100

88.7–100

5a

9.6 (50)

120

118

0

2

100

96.9–100

6a

9.6 (50)

120

114

3

3

97.4

92.7–99.1

6a

14.4 (75)

30

30

0

0

100

88.7–100

* Confidence Interval

Note: Invalid results were not included in the data calculations for Percent Valids Detected.

Clinical specimens representing HCV genotypes 1 to 6 at different concentrations were used to determine the percent detection of the Aptima HCV RNA qualitative assay. The specimens were quantitated using the VERSANT HCV RNA 3.0 assay (bDNA). The genotypes of the specimens were provided by the specimen vendor and confirmed using the VERSANT HCV Genotype assay (LiPA) and sequencing. The results are shown in Table 3.The overall percent detected across all HCV genotypes tested was ≥95% at 9.6 IU/mL (50 copies/mL).

Please see package insert for detailed information.

 

 

Regulatory

The Aptima HCV RNA qualitative assay was FDA-approved for in vitro diagnostic use in 2006. It is not FDA-approved for screening of blood or plasma donors.

For additional availability in other countries outside of the U.S., please contact your local sales representative or distributor.

References
  1. www.cdc.gov/hepatitis/HCV/PDFs/HepCTesting-Diagnosis.pdf.
  2. Current and Future Disease Progression of the Chronic HCV Population in the United States (2013) PLOS ONE Volume 8: Issue 5; 1-10.
  3. http://www.cdc.gov/KnowMoreHepatitis/.
  4. Treatment of Hepatitis C, A Systemic Review (2014) Volume 312: No.6; 631-640.
  5. Aptima HCV RNA Qualitative Assay Package Insert. San Diego, CA: Gen-Probe Incorporated, 2011.