CMV Dried Blood Spot Testing
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1 CMV Dried Blood Spot Testing Michael Boeckh, MD Member, Vaccine and Infectious Disease Division Head, Infectious Disease Sciences Program Fred Hutchinson Cancer Research Center Professor, Department of Medicine, University of Washington Seattle, Washington, USA
2 Late CMV Disease Continuing Challenge Who is at Risk? CMV serostatus Covariates Multivariable Adjusted HR (95% CI) D-/R+ 1 P value D+/R ( ) D+/R ( ) Acute GVHD grade ( ) CMV viremia before day ( ) CMV disease before day ( ) Lymphocyt e count at day cells/mm 3 1 < 300 cells/mm ( ) Missing 0.90 ( ) Corticosteroid treatment after day ( ) <0.001 CMV Viremia after day ( ) <0.001 Ōtzkōk et al. ASH 2014, Tandem 2015
3 Surveillance Adherence Özkök S et al. ASH 2014
4
5 Late CMV Disease: Challenges
6 Late CMV Disease: Challenges
7 Patient-based Testing Limaye et al. JCM 2013
8 Assay Evolution DESCRIPTION Initial Procedure (Limaye et al. 2013) CURRENT PROCEDURE CMV On The Go Filter paper used Whatman FTA Elute Whatman 903 Protein cards Saver cards Size of each punch 3 mm in diameter 6 mm in diameter Number of punchs used 4 2 for DNA extraction DNA extraction procedure PCR master mix PCR Machine IU conversion Internal control template Internal control primers and probe washed the DBS twice with 1 ml of water, then added 100 ul of 5% Chelex and incubated the vials at 95 C for 30 minutes to elute the DNA from the paper QuantiTect Muliplex PCR mix with ROX 7900HT sequence detection system 1 IU= 4 copies (AcroMetrix)* Spiked into PCR master mix EXO pre-treat DBS with proteinase K, spike intermal control template, perform DNA extraction using Promega Maxwell 16 Viral Total Nucleic Acid Purification Kit on Promega Maxwell 16 MDx Instrument. QuantiTect Muliplex PCR NoROX mix QuantStudio 7 flex real-time PCR system 1 IU=1.4 copies (WHO) Spike into extraction lysis buffer EXO BS
9 CMV on the Go Left-over blood from clinical testing (UW/SCCA)
10 Proposed trial for DBS CMV testing after day 100 Day 100 Fred Hutchinson Cancer Research Center 10
11 PATIENT OR Local Oncologist My CMV Portal hosted by Emmes Advantage eclincal Molecular Diagnostics Lab Bi-directional Communication Bi-directional Communication with Password Protected Secure Access Uni-directional Communication Uni-directional Communication with Password Protected Secure Access
12 PATIENT VERSION 1 PATIENT Emmes Advantage eclincal Local Oncologist My CMV Tests Patient Portal Uni-directional Communication Bi-directional Communication Password Protected Secure Access TBD Molecular Diagnostics Lab Last updated: 10/10/2016
13 STUDY SITES VERSION 1 PATIENT Emmes Advantage eclincal Local Oncologist My CMV Tests Patient Portal Uni-directional Communication Bi-directional Communication Password Protected Secure Access TBD Molecular Diagnostics Lab Last updated: 10/10/2016
14 LABORATORY VERSION 1 PATIENT Emmes Advantage eclincal Local Oncologist Uni-directional Communication Bi-directional Communication Password Protected Secure Access TBD Molecular Diagnostics Lab Last updated: 10/10/2016
15 PRIMARY TREATING PHYSICIAN VERSION 1 PATIENT Local Oncologist Uni-directional Communication Bi-directional Communication Password Protected Secure Access TBD Molecular Diagnostics Lab Last updated: 10/10/2016
16 Emmes VERSION 1 PATIENT Emmes Advantage eclincal Uni-directional Communication Bi-directional Communication Password Protected Secure Access TBD Molecular Diagnostics Lab Last updated: 10/10/2016
17 Testing Procedure
18 Acknowledgments Ajit Limaye Margaret Green Louise Kimball Cindy Hirano James Reith Lisa Chung Hu Xie Wendy Leisenring Amalia Margaret Zach Stednick MeeiLi Huang Linda Cook Keith Jerome
19 Schematic of Study Design Prior to Enrollment N = 150: Obtain informed consent; assess eligibility per criteria Initial Study Visit Randomize subjects; confirm CMV monitoring interval; counsel re: adherence; train in DBS collection; train in website/app use Weeks 1-13 DBS self-collection First Quarterly Contact (Week 13) CMV disease monitoring; safety monitoring; adherence counseling Weeks DBS self-collection Second Quarterly Contact (Week 26) Weeks Third Quarterly Contact (Week 39) As needed; subject duration on study will vary CMV disease monitoring; safety monitoring; adherence counseling DBS self-collection CMV disease monitoring; safety monitoring; adherence counseling Non-Randomized Controls - Medical records review at 365d post-transplant for survival and CMV disease Weeks DBS self-collection Closeout Contact CMV disease monitoring; safety monitoring Assessment of Final Study Outcome Measures
Disclosures. Investigator-initiated study funded by Astellas
Disclosures Investigator-initiated study funded by Astellas 1 Background Widespread use of preemptive therapy strategies has decreased CMV end-organ disease to 5-8% after HCT. Implications for development
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