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Holland Laboratory David A. Leiby, PhD Transmissible Diseases Department American Red Cross Holland Laboratory and Department of Microbiology and Tropical.

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Presentation on theme: "Holland Laboratory David A. Leiby, PhD Transmissible Diseases Department American Red Cross Holland Laboratory and Department of Microbiology and Tropical."— Presentation transcript:

1 Holland Laboratory David A. Leiby, PhD Transmissible Diseases Department American Red Cross Holland Laboratory and Department of Microbiology and Tropical Medicine George Washington University Experience With Testing Blood Donors for Babesia

2 Outline B. microti seroprevalence studies 2009 NAT pilot study Longitudinal natural history study Lookback results

3 B. microti Seroprevalence Study  conducted from 1999 to present  primarily focused in Connecticut  recently expanded to include MA, NH & ME  donors tested by IFA for antibodies to B. microti  cutoff titer of 1:64  IFA positive donors deferred indefinitely  during first 2 years, deferred based on PCR +  studies initially targeted highly endemic areas, but subsequently expanded statewide

4 CT Donor Seroprevalence: 1999-2007 Year # Tested Antibody + (%)PCR + (%) 1999 3,490 30 (0.9)10/19 (53) 2000 2,681 28 (1.0)10/18 (56) 2001 2,162 30 (1.4) 2/25 (8) 2002 2,230 18 (0.8) 2/14 (14) 2003 1,989 34 (1.7) 2/20 (10) 2004 2,864 43 (1.5) 1/33 (3) 2005 1,841 25 (1.4) 0/10 (0) 2006 3,251 39 (0.9) 3/19 (16) 2007 5,267 42 (0.8) 1/22 (5)

5 County Results: 2000-2008 County# IFA Positive/# IFA Test (% Seropositive) Fairfield17/3737 (0.5) Hartford26/4690 (0.6) Litchfield6/1616 (0.4) Middlesex43/3167 (1.4) New Haven15/2907 (0.5) New London161/9634 (1.7) Tolland7/787 (0.9) Windham7/1040 (0.7)

6 Spatial cluster 1 Spatial cluster 2 Seroprevalence/10,000 donations Johnson et al., Transfusion 2009;49:2574-2582

7 Piloting NAT for B. microti  pilot study of ~1,000 CT donations  collected August/October 2009 from Middlesex and New London Counties  1,002 tested to date:  25 (2.5%) IFA positive  3 (0.3%) PCR positive (2 IFA +, 1 IFA -)  all identified by first week of September  1 apparent window period infection detected  number likely low  acutely infected donors too sick to donate?  role for NAT during tick season?

8 Babesia NAT Approach  seasonally triggered  May through September  targets acute or “window period” infections  technologic hurdles remain:  PCR sensitivity sufficient, but...  parasitemia low compared to viral infections  requires whole blood  limited volume for testing  considerations of concentration techniques

9 Babesia Natural History Study  long-term, ongoing study in CT and MA  enrolling B. microti seropositive donors from seroprevalence study  tested every 30 - 60 days  serology  blood smear  PCR  hamster inoculation (w/ CDC)  risk-factor questionnaires  initial risk  potential re-exposure  investigate infection/resolution patterns over time

10 Natural History Study Observations  227 seropositive donors enrolled to date  184 evaluable donors from 2000–2008  86 (47%) sero-reverted to baseline negative  98 (53%) remained seropositive  37 (38%) resolving infection  30 (31%) chronic infections  12 (12%) re-infections  19 (19%) insufficient data

11 Resolution of Babesia Infection: Part I Date IFA PCR Hamster 07/21/00 1:512 ND ND 08/15/00 1:256 Pos. Pos. 09/18/001:128 Pos. Neg. 12/01/00 1:128 Neg. Neg. 04/06/01 <1:64 Neg. Neg. 07/13/01 <1:64 Neg. Neg. 10/12/01 <1:64 Neg. Neg. Released from study

12 Western Blot Analysis

13 Resolution of Babesia Infection: Part II Date IFA PCR Hamster 07/28/03 1:128 ND ND 08/25/03 1:128 Neg. Neg. 10/17/03 1:64 Neg. Neg. 03/16/04 1:64 Neg. Neg. 07/01/04 <1:64 Neg. Neg. 10/01/04 <1:64 Neg. Neg. Released from study

14 Date IFA PCRHamster 08/24/00 1:512 ND ND 10/02/00 1:512 Pos. Pos. 11/28/00 1:512 Neg. Pos. 11/30/00 initiated 10 day treatment for babesiosis 03/30/01 1:512 Neg. Neg. 05/31/01 1:512 Neg. Neg. 09/10/01 1:256 Neg. Neg. 12/01/01 1:512 Neg. Neg. 02/25/02 1:512 Neg. Neg. 07/18/03 1:512 Neg. Neg. Released from study Persistent High Ab Titer = Chronic Carrier

15 Western Blot Analysis

16 DateIFA PCR Hamster 07/07/00 1:64 ND ND 08/03/00 1:128 Pos. Pos. 09/21/00 1:128 Pos. Neg. 11/08/00 1:256 Neg. Neg. 02/15/01 1:128 Neg. Neg. 04/28/01 1:64 Neg. Neg. 08/17/01 1:128 Neg. Neg. 10/26/01 1:64 Neg. Neg. 12/21/01 <1:64 Neg. Neg. 02/22/02 <1:64 Neg. Neg. 05/02/02 1:256 Neg. Neg. 07/12/02 1:64 Neg. Neg. 10/08/02 1:128 Neg. Neg. 12/13/021:64 Neg. Neg. 02/21/031:64 Neg. Neg. 05/12/031:128 Neg. Neg. 07/23/031:128 Neg. Neg. Re-exposure to B. microti?

17 Western Blot Analysis

18 Date IFA PCR RT-PCR Hamster 07/31/031:1024 ND ND ND 08/26/03** 1:1024 Pos. Pos. Pos. 08/29/03 initiated 10 day treatment for babesiosis 09/22/03 1:1024 Neg. Neg. Neg. 10/23/03 1:256 Neg. Pos. Neg. 01/02/04 1:1024 Neg. Pos. Neg. 03/16/04 1:1024 Pos. Pos. Neg. 04/27/041:1024 Neg. Pos. Neg. 06/26/041:1024 Neg. Pos. Neg. 09/23/04 1:1024 Neg. Pos. Neg. 11/29/04 1:128 Neg. Neg. Neg. 01/21/05 1:1024 Neg. Neg. Neg. 03/28/05 1:512 Neg. Neg. Neg. 05/17/05 1:512 Neg. Pos. Neg. 07/12/05 1:256 Neg. Pos. Neg. 09/19/05 1:256 Neg. Neg. Neg. 12/02/05 1:512 Neg. Neg. Neg. 03/07/06 1:512 Neg. Pos. Neg. 06/28/06 1:512 Neg. Neg. Neg. * 79 year-old male ** positive on blood smear Asymptomatic Chronic Carrier

19 Chronic Carrier – NIH Blood Donor  donor in the NIH blood program  implicated in a transfusion case  asymptomatic chronic carrier  hereditary hemochromatosis patient  frequent marathon runner  excellent health  implicated donation  Ab titer of 1:1024  PCR negative  follow-up studies  implicated in additional transfusion case  Ab titers remain at 1:1024 (n=11)  never PCR positive

20 Year IFA Positive Donors (%) Lookback Donations Lookback On Cellular Products Recipients Tested Lookbacks Positive (%) 199930/3669 (0.8%)14519482 200028/2681 (1.0%)8110393 200130/2162 (1.4%)325051 200218/2230 (0.8%)385882 200334/1989 (1.7%)518440 200443/2864 (1.5%)83113170 200525/1841 (1.4%)4454120 Total208/17436 (1.2%)474656638 (12.7%) CT Lookback Results (1999-2005)

21 Summary  seroprevalence among CT donors ~ 1% each year  portion of donors parasitemic  focal endemicity, but statewide distribution  NAT role indicated for seasonal, acute infections  natural history of babesial infection  majority of donors resolve infection  considerations for re-entry  chronic carriers pose significant transmission risk  lookback investigations verify transmission risk

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