National Screening laboratory Sanquin The use of NAT in blood donation screening in The Netherlands. TransMed conference Bhopal 18 November 2016

Transcription

1 Dr. Anton van Weert

2 National Screening laboratory Sanquin The use of NAT in blood donation screening in The Netherlands TransMed conference Bhopal 18

3 Introduction Thank you! Anton van Weert, PhD, MBA Manager Donorscreening and Infectious Disease Diagnostics Division Sanquin Diagnostic Services National Screening laboratory Sanquin The Netherlands Tel:

4 Agenda Introduction to Sanquin Blood supply Donorscreening in The Netherlands - logistics and IT - NAT-screening Validation and use of the S201 molecular platform for high throughput testing of pooled donations 4

5 Sanquin Blood Supply Foundation

6 The Netherlands Number of inhabitants: Surface area: 41,528 km 2 Number of hospitals: 133 6

7 Sanquin Amsterdam HQ

8 More than just blood Blood collection Recruiting and medically examining donors Collecting blood and plasma Products and supply Processing and testing blood and blood products Stock management Supplying products (blood, plasma, medicines, reagents) and services to hospitals and other customers Education and advice Advising customers about products and services Performing scientific research Carrying out diagnostics

9 Organisation Executive Board Corporate staff Blood Bank Plasma Products Ltd. Diagnostic Services Research Reagents Tissues & Cells (business unit) Diagnostic testing of patients Product controls Screening blood donations Development new screening applications Validation of diagnostic tests and analyzers

10 Blood supply in The Netherlands Main collection locations: Amsterdam and Nijmegen Collection points: - 56 permanent mobile

11 Key figures blood supply 2015 (1)

12 Key figures blood supply 2015 (2) Blood products Use of red blood cell concentrates 427,685 Platelets (from whole blood in donor units) 251,625 Fresh frozen plasma Kilograms plasma total (incl. apheresis) supplied to Plasma Products division 7, ,404

13 Key figures blood supply 2015 (3) Positive donations HBV HCV HIV Lues New donors repeat donors New donors repeat donors New donors repeat donors New donors repeat donors

14 National Screening laboratory Sanquin (NSS)

15 National Screening laboratory Sanquin (NSS) Pre-analysis

16 National Screening laboratory Sanquin (NSS) Virus serology Blood group serology

17 National Screening laboratory Sanquin (NSS) NAT infectious diseases

18 Logistics & IT

19 Flow of data (Re)testing required Searching samples Lab results LIMS Authorization day 1 after whole blood collection before 14:00 Release components eprogesa

20 Logistics Whole blood Apheresis platelets Apheresis plasma New blood donor First blood donation

21 Logistics ( ) Tue - Fri: ~3,250 Sat: ~1,800 New donors: ~8%

22 Policy regarding screening results Screening negative: eligible Screening positive: rejected Screening reactive: donation destroyed confirmation negative: donor may donate again Screening 2nd (=next) donation reactive: donation destroyed confirmation negative: donor excluded for 2 years After exclusion for 2 years: enrollment as a new donor; requirement: blood group must have been confirmed 22

23 Blood Donation Testing

24 Blood group typing ABO RhD C,c,E,e,K a-tp Phenotyping other RBC-antigens Olympus PK7300 (4x)

25 Serology HBsAg a-hbcag a-hiv-1/2 a-hcv (a-htlv-i/ii on the Architect) PRISM Next (5x)

26 Additonal analyses Automatic search and selection tubes using the PVT: - anti-parvo B19 - anti-malaria - Total Protein - anti-coxiella burnetii

27 NAT-screening 2008: S201 Single target - HBV DNA - HCV RNA - HIV RNA Pools of 6 Hamilton (4x) 2013: S201 MPX v2.0 Ampliprep Taqman (8x)

28 NAT capacity planning Workflow analysis Sanquin and Roche # donations Hour Delivery of test tubes NAT test results available

29 S201 validation and performance

30 S201: validation (1) Acceptation criteria - % invalid pools due to pipetting errors Hamilton <1% - % invalid pools due to pipetting errors AmpliPrep/TaqMan <1% - % initial reactive pools <0.5% - % rejected batches <5% - results 1200 donations available <12 hours - dilutions international standards HIV RNA, HCV RNA and HBV DNA must be reactive at a concentration of 3x detection limit - No cross contamination from positive to negative samples Performancy Qualification - System must work without problems - All results must be valid

31 S201: validation (2) General - Two lots of the MPX kits - Two lots of the control kits - Validation experiments conducted by 5 technicians - Testing on two AmpliPrep/TaqMan systems Cross contamination - Samples with HBV DNA >10 IU/mL among negative samples - test of the entire analysis chain: sample preparation, de-capping, pooling on the Hamilton and subsequent testing

32 S201: validation (3) Hamilton Star Lab Number/% Remarks Donations (negative) Pipetting errors 5 % pipetting errors 0.03% AmpliPrep/TaqMan Number/% Pools 2708 Invalid pools 2 % invalid pools 0.07% sample error (3); too less sample volume (1); unknown (1) No/too low IC signal (2)

33 S201: validation (4) Specificity TaqScreen MPX test: Categorie Number % Pools tested non-reactive pools Initial reactive pools Non-repeated reactives (negative)

34 S201: validation (5) Sensitivity TaqScreen MPX test: Target signal (cycles) IS WHO Concentration Ct-value St. Dev. 3x LOD (11.1 IU/mL) HBV (n=16) 6x LOD (22.2 IU/mL) HCV (n=16) HIV (n=16) 3x LOD (32.1 IU/mL) x LOD (64.2 IU/mL) x LOD (147 IU/mL) x LOD (294 IU/mL) All 16 out of 16 samples were positive for all 3 viruses

35 S201: validation (6) Sensitivity TaqScreen MPX test: external positive control: Target signal (cycles) PeliSpy Pro Concentration Concentration (IU/mL) Ct-value St. Dev. HBV (n=5) x LOD HCV (n=5) x LOD HIV-1 (n=5) 250 5x LOD

36 S201: validation conclusions The cobas TaqScreen MPX test for the S201 system meets the acceptance criteria defined prior to the validation The cobas TaqScreen MPX test can be implemented for screening of donations for the presence of HBV, HCV and HIV The S201 systems can be released for routine donor screening The server of result management and the back-up of this server work as expected

37 S201: operational process Pooling on Hamilton: - total volume of 1 ml per 6 donations Prepare batches of max 24 samples: - max 20 pools of 6 donations - 4 samples from the kit control Every day also test UDEC (User Defined External Control) Reactive pools: resolution testing individual donations Repeat reactive: confirmation testing

38 S201 next step: MPX 2.0 implementation 2013 implemented - CE-marked system - Evaluation in other countries completed Performancy Qualification - robustness - reproducibility

39 Performance MPX2.0 test on s201 system MPX2.0 Validation 2012 MPX2.0 routine screening Number donations 3,312 2,078,592 Number of batches with pools ,652 Number of pools ,432 Invalid batch rate 3.6% 2.6% (1% - 6.1%) Invalid test rate* 0.92% 0.44% False positive rate pools % 0.03% (0% -0.14%) Specificity on pools % 99.97% False positive donations** 0 1 (HIV)

40 Performance MPX2.0 test on s201 system MPX2.0 validation 2012 MPX2.0 routine screening 04/ /2016 Number donations 3,312 2,078,592 Number of batches with pools ,652 Number of pools ,432 Invalid batch rate 3.6% 2.6% (1% - 6.1%) Invalid test rate* 0.92% 0.44% False positive rate pools % 0.03% (0% -0.14%) Specificity on pools % 99.97% False positive donations** 0 1 (HIV) * Invalids tests due to lack of IC signal and math errors **repeat testing source-donation samples, archive samples and new donations were all HIV-negative

41 Concluding remarks India Amsterdam The Netherlands

42 Questions Anton van Weert: