# Presentation: 197980

## COBAS AmpliScreen HBV Test HBV Seroconversion Panel Results and HBV NAT Positive / Serology Negative Donors Larry PietrelliDirector, Clinical AffairsRoche Molecular Diagnostics

## Data Summary

- Today’s focus:
- Overview of the COBAS AmpliScreen HBV Test Performance on 40 HBV Seroconversion Panels Compared to HBsAg Tests
- Clinical Study Summary of Donors Found HBV DNA Positive / HBsAg & anti-HBc Negative &  Enrolled in Follow-up Studies

## Seroconversion Panel TestsDetection of HBV DNA Prior to Ortho HBsAg Test System 3

- 	On average, the COBAS AmpliScreen HBV Test detected HBV **15** days with Mini-pool Test and **20** days with Single Unit Test earlier than the Ortho HBsAg Test System 3. There was no case in which serological tests detected HBV prior to NAT.
- .

## Seroconversion Panel TestsPanel #24 Detailed Test Results

| Panel member | Days since 1st Bleed | Days from 1st Single NAT + | ORTHO HBsAg ELISA Test System 3SCO ≥1.0 = + | Mini-Pool Test
Abs. ≥ 0.2 = +  | Single Unit Test Abs. ≥ 0.2 = + |
| --- | --- | --- | --- | --- | --- |
| 1 | 0 | NA | 0.29 | 0.005 | 0.004 |
| 2 | 2 | NA | 0.10 | 0.005 | 0.003 |
| 3 | 9 | NA | .038 | 0.004 | 0.006 |
| 4 | 11 | NA | 0.18 | 0.004 | 0.003 |
| 5 | 16 | 0 | 0.01 | 0.005 | 3.961 |
| 6 | 18 | 2 | 0.01 | 3.968 | 3.961 |
| 7 | 31 | 13 | 1.22 | 3.968 | 3.660 |
| 8 | 36 | 20 | 2.78 | 3.968 | 3.660 |
| 9 | 45 | 29 | 37.58 | 3.491 | 3.660 |
| 10 | 49 | 33 | 65.94 | 3.667 | 3.484 |

## Seroconversion Panel TestsPanel #40 Detailed Test Results

| Panel member | Days since 1st Bleed | Days from 1st Single NAT +
 | ORTHO HBsAg ELISA Test System 3SCO ≥1.0 = + | Mini-Pool Test
Abs. ≥ 0.2 = +  | Single Unit Test Abs. ≥ 0.2 = + |
| --- | --- | --- | --- | --- | --- |
| 1 - 13 | 0 - 53 | 0 - 53 | < 1.0 | > 0.2 | > 0.2 |
| 14 | 58 | 58 | 0.12 | 4.000 | 4.000 |
| 15 | 60 | 60 | 0.20 | 3.767 | 4.000 |
| 16 | 65 | 65 | 0.06 | 4.000 | 4.000 |
| 17 | 67 | 67 | 0.01 | 4.000 | 4.000 |
| 18 | 72 | 72 | 0.01 | 4.000 | 4.000 |
| 19 | 74 | 74 | 0.13 | 4.000 | 3.537 |
| 20 | 94 | 94 | 0.12 | 4.000 | 4.000 |
| 21 | 97 | 97 | 0.20 | 4.000 | 4.000 |
| 22 | 101 | 101 | 0.41 | 4.000 | 4.000 |
| 23 | 104 | 104 | 0.99 | 4.000 | 4.000 |
| 24 | 108 | 108 | 1.10 | 4.000 | 4.000 |
| 25 | 111 | 111 | 2.84 | 3.827 | 4.000 |
| 26 | 115 | 115 | 4.32 | 4.000 | 4.000 |

## Seroconversion Panel TestsPanel #39 Detailed Test Results

| Panel member | Days since 1st Bleed | Days from 1st Single NAT + | ORTHO HBsAg ELISA Test System 3SCO ≥1.0 = + | Mini-Pool Test
Abs. ≥ 0.2 = +  | Single Unit Test Abs. ≥ 0.2 = + |
| --- | --- | --- | --- | --- | --- |
| 1 | 0 | 0 | 0.07 | 3.959 | 3.959 |
| 2 | 2 | 2 | 0.4 | 0.002 | 3.955 |
| 3 | 42 | 42 | 0.07 | 3.959 | 3.959 |
| 4 | 73 | 73 | 0.13 | 0.003 | 3.959 |
| 5 | 91 | 91 | 0.04 | 0.004 | 0.002 |
| 6 | 96 | 96 | 0.13 | 0.003 | 0.003 |
| 7 | 98 | 98 | -0.16 | 0.004 | 3.959 |
| 8 | 103 | 103 | 0.1 | 0.005 | 0.002 |
| 9 | 106 | 106 | -0.01 | 0.005 | 1.662 |
| 10 | 111 | 111 | 0.43 | 0.016 | 3.482 |
| 11 | 113 | 113 | 0.25 | 3.959 | 0.003 |
| 12 | 143 | 143 | 34.98 | 3.482 | 2.637 |
| 13 | 148 | 148 | 81.08 | 3.357 | 2.561 |
| 14 | 150 | 150 | OVER | 3.658 | 2.512 |

## Summary of COBAS AmpliScreen HBV Test vs. Ortho HBsAg ELISA Test System 3 with 39 HBV Seroconversion Panels

** **One seroconversion panel was not included in the calculations which was detected by the COBAS AmpliScreen HBV Test 108+ days prior to the Ortho HBsAg ELISA Test System 3. *

|  | Mini-Pool Test | Single Unit Test |
| --- | --- | --- |
| Mean | 15 | 20 |
| Median | 14 | 18 |
| Maximum | 30 | 53 |
| Minimum | 0 | 0 |

## Clinical Trial Summary

| Results | Total |
| --- | --- |
| Anti-HBc - / HBsAg - / DNA - | 578,671 |
| Anti-HBc + / HBsAg + / DNA + | 84 |
| Anti-HBc + / HBsAg + / DNA - | 16 |
| Anti-HBc + / HBsAg - / DNA - | 2,988 |
| Anti-HBc - / HBsAg + / DNA + | 3 |
| Anti-HBc - / HBsAg - / DNA + | 23 |
| Anti-HBc + / HBsAg - / DNA + | 1 |
| Anti-HBc - / HBsAg + / DNA - | 4 |
| Total | 581,790 |

## Follow-up Study

- Donor enrolled in the follow-up study were to return weekly for the first month and then monthly for 5 months for a total of 6 months
- Testing was to include the following:
- IgM anti-HBc
- anti-HBc (total)
- anti-HBs
- HBsAg
- HBV DNA

## Potential Window Cases

- 23 Donors were HBV DNA + / HBsAg - / Anti-HBc –
- 	- 14 were enrolled into the follow-up study
- 	- 2 confirmed Window Period cases
- 12 presumed false positive due to persistently negative Anti-HBc, HBsAg and HBV DNA
- 9 subject declined follow-up (calculated as false positive for sensitivity/specificity determination)

## Follow-up Subject AA110001

| Day | Anti-HBc | HBsAg | HBV DNA | Anti-HBs | IgM Anti-HBc | Quant. cps/mL |
| --- | --- | --- | --- | --- | --- | --- |
| Index | NR | NR | Positive | ND | ND | 2,000 |
| 17 | NR | Positive | Positive | NR | NR | 120,000 |
| 40 | NR | Positive | Positive | NR | NR | 3.1E9 |
| 48 | RR | Positive | Positive | NR | NR | 3.5E9 |
| 55 | RR | Positive | Positive | NR | RR | 4.7E8 |
| 202 | RR | Positive | Positive | NR | RR | >5.0E9 |

- 26 year old male repeat donor with no know risk factors

## Follow-up Subject DA120001

| Day | Anti-HBc | HBsAg | HBV DNA | Anti-HBs titer | IgM Anti-HBc | Quant. cps/mL |
| --- | --- | --- | --- | --- | --- | --- |
| Index | NR | NR | Positive | 2340 | ND | 200 |
| 8 | NR | NR | Positive | 1980 | NR | 700 |
| 22 | RR | NR | Positive | 2730 | NR | 200 |
| 29 | RR | NR | Negative | 3350 | RR | 100 |
| 57 | RR | NR | Negative | 112 | RR | ND |
| 168 | NR | NR | Negative | 55 | RR | ND |

- 49 year old female, repeat donor, health care worker, previously vaccinated

## Additional Window Cases Detected Post Clinical Study

- Three of the five sites continued screening for HBV under the IND after conclusion of the Clinical Study
- Three additional window period cases identified

## Follow-up Subject LA150001

| Day | Anti-HBc | HBsAg | HBV DNA | Anti-HBs | IgM Anti-HBc | Quant. cps/mL |
| --- | --- | --- | --- | --- | --- | --- |
| Index | NR | NR | Positive | ND | ND | 61,000 |
| 7 | NR | Positive | Positive | RR | NR | ND |
| 26 | RR | NR | Negative | RR | RR | ND |
| 54 | RR | NR | Negative | RR | RR | ND |

- 27 year old male repeat donor, multiple high-risk male sexual partners

## Follow-up Subject LA150002

| Day | Anti-HBc | HBsAg | HBV DNA | Anti-HBs | IgM Anti-HBc | Quant. cps/mL |
| --- | --- | --- | --- | --- | --- | --- |
| Index | NR | NR | Positive | ND | ND | 2,300 |
| 7 | NR | Positive | Positive | NR | NR | ND |
| 14 | NR | Positive | Positive | NR | NR | ND |
| 21 | NR | Positive | Positive | NR | NR | ND |
| 28 | RR | NR | Positive | NR | RR | ND |
| 63 | RR | NR | Negative | RR | RR | ND |

- 29 year old male repeat donor, only possible risk factor was acupuncture once a week for 8 weeks prior to donation

## Follow-up Subject LA151303

| Day | Anti-HBc | HBsAg | HBV DNA | Anti-HBs | IgM Anti-HBc | Quant. cps/mL |
| --- | --- | --- | --- | --- | --- | --- |
| Index | NR | NR | Positive | ND | ND | 37,000 |
| 14 | NR | Positive | Positive | NR | NR | ND |

- 50 year old male repeat donor

## Conclusions from the Window Period Cases Identified

- Four of the five window period cases seroconverted to HBsAg positivity.  Time from first minipool NAT positivity to HBsAg Positivity Ranged from 7 to 17 with a median of 11 days.
- One donor who was previously vaccinated against HBV failed to seroconvert.
- 12 donors were negative for all markers at the end of the 6 month follow-up period and were classified as false positive