Quality Assurance Program for Cystic Fibrosis Newborn Screening
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Marie C. Earley, Ph.D. Research Microbiologist APHL Training Course June 29, 2011 Quality Assurance Program for Cystic Fibrosis Newborn Screening National Center for Environmental Health Newborn Screening Quality Assurance Program
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Quality Assurance Program for Cystic Fibrosis Newborn Screening. Marie C. Earley, Ph.D. Research Microbiologist APHL Training Course June 29, 2011. National Center for Environmental Health. Newborn Screening Quality Assurance Program. - PowerPoint PPT Presentation
Transcript of Quality Assurance Program for Cystic Fibrosis Newborn Screening
Marie C. Earley, Ph.D.Research Microbiologist
APHL Training CourseJune 29, 2011
Quality Assurance Program for Cystic Fibrosis Newborn Screening
National Center for Environmental HealthNewborn Screening Quality Assurance Program
Quality assurance for laboratories involved in screening newborns and children for heritable disorders
Quality assurance for newborn-screening tests
Performance evaluation services Technical assistance and technology
transfer to newborn screening laboratories
Assistance to ensure analytic validity and utility of screening tests
NSQAP has been mandated by Congress to provide QA materials
for NBS Laboratories
Newborn Screening Saves Lives Act of 2008
NSQAP shall provide for:
Appropriate quality control and other performance test materials to evaluate the performance of new screening tools
The Newborn ScreeningQuality Assurance Program
The only comprehensive quality assurance program using the dried-
blood spots100% participation of US states 456 labs reported data
67 countries participated 391 labs participated in PT
717,255 DBS produced 337 labs participated in QC
28 employees 17 reports to participants
36 new enrollments 4 filter paper lots evaluated
463 labs enrolled at year end
CF Mutation Detection Proficiency Testing
Program
CF Mutation Detection Pilot Proficiency Testing Program
Began as a collaborative effort between CDC and 3 CF Centers
Specimens drawn from adult or adolescent CF patients and are NOT enriched with IRT (No IRT testing done).
Began quarterly shipments in February 2007
Program has grown from 25 to 60 laboratories
Repository contains all of the ACMG recommended mutations and additional rare mutations
Many Different Methods Luminex Diagnostics (Luminex Platform) Hologic Invader Assay (Invader) Innogenetics Inno-Lipa (Hybridization) Abbott Diagnostics Oligonucleotide
Ligation Assay Roche Diagnostics Linear Array
(Hybridization)* Genprobe Diagnostics Elucigene (4
different kits, ARMS) MALDI-TOF mass spectrometry In-house (TaqMan, SNP, hydolysis probe,
Lightcycler) Home brew Amplification/gel electrophoresis Amplification/Heteroduplex/restriction
analysis Sequencing
QuarterSpecimens
Assayed (N)a
Correct Assessments
N (%)Transcription Errorc N (%)
True Miss N (%)
20071 77 76 (99.7) 0 (0.0) 0 (0.0)2 75 75 (100) 0 (0.0) 0 (0.0)3 94 91 (96.8) 0 (0.0) 0 (0.0)4 103 94 (91.3) 1 (1) 0 (0.0)
20081 107 104 (97.2) 2 (2.0) 0 (0.0)2 124 120 (96.8) 0 (0.0) 1 (0.8)3 142 138 (97.2) 1 (0.7) 0 (0.0)4 182 175 (96.2) 6 (3.3) 0 (0.0)
20091 181 179 (98.9) 0 (0.0) 1 (0.6)2 175 174 (99.4) 0 (0.0) 0 (0.0)3 161 160 (99.4) 0 (0.0) 1 (0.6)4 204 202 (99.0) 0 (0.0) 1 (0.5)
Total 1625 1588 (97.7) 10 (0.6) 4 (0.2)
Incorrect assessments potentially leading to a missed
case (%)b
Proficiency testing results summary
Quality Control Materials
Making CF QC Materials for Molecular Methods
Wash leuko-
reduced blood with saline (3X)
Add filtered serum
Adjust hematocrit
Add lymphocytes
Spot onto paper
Analyze and store at -20C
Laboratory-Created Molecular QC Materials
CFTR Mutation Analysis QA materials created from
transformed cell lines Six pools from an individual cell line were
evaluated in-house Six pools evaluated externally by CF Mutation
Detection PT Program participants Concentration of cells has been set
Five more pools using cell lines with different mutations have been prepared
Future materials will include additional CFTR mutations using the optimized protocol
This protocol will be expanded to future disorders as needed
Cystic Fibrosis Quality Control Materials
First set of specimens sent as educational specimens (no grading of results)
All were made with an F508del carrier cell line
96% response rate Most labs could pick up , a few couldn’t and
it was a small subset of the entire group Continue with evaluations before sending
out as pilot QC materials We are working towards having the ACMG
panel covered by the end of 2011 We will continue to find a way to have cell
lines for other mutations commonly found in commercial kits
Other projects
Ongoing collaboration with Ghana to establish a mutually beneficial program to expand NBS proficiency testing for hemoglobinopathies
CDC will characterize the specimens and include them as PT specimens
Why Ghana? Ghana has one of the highest incidences of sickle cell
disease worldwide
Hemoglobinopathies Proficiency Testing
Country Population (millions)
Birth Rate
Total Births SCD Birth Rate
Total SCD Births
Ghana 23.8 29.6 705,575 1:55 12,829USA 308.8 14.0 4,323,634 1:2,474 1,748
Characterization of Hemoglobinopathy DBS
After specimens are received and logged, each specimen will be tested by: Microsatellite repeats to test for contamination Isoelectric focusing High Performance Liquid Chromatography Sequencing of HBB1, HBA1, and HBA2 genes Deletion analysis, if necessary Mass spectrometry
Complete characterization will provide assurance that the variants are known and will provide data for comparing the different methods used for analysis
Conclusions NSQAP is working to develop and sustain PT
programs for molecular assays used in NBS The CF Mutation Detection Program has
been a success and NBS labs have shown that they can successfully perform molecular assays
Pilot QC materials for CF molecular assays look promising and we are moving forward with plans to expand the number of mutations available for QC
We are moving forward to increase the variety of hemoglobinopathy specimens
Acknowledgements
Case Western Reserve University
Dr. Michael KonstanKate Hilliard
The Johns Hopkins Hospital
Dr. Peter Mogayzel
All of our CF donors and NBS PT participants
Centers for Disease Control and Prevention
Newborn Screening and Molecular Biology
Branch
University of Wisconsin
Dr. Phil FarrellAnita Laxova
For more information please contact Centers for Disease Control and Prevention1600 Clifton Road NE, Atlanta, GA 30333Telephone, 1-800-CDC-INFO (232-4636)/TTY: 1-888-232-6348E-mail: [email protected] Web: www.cdc.govThe findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention.
National Center of Environmental HealthNewborn Screening Quality Assurance Program
Newborn Screening & Molecular Biology Branch
Dana ChafinSuzanne Cordovado, PhDMiyono HendrixSean MochalJoanne Mei, PhDCarla Cuthbert, PhDNSQAP Administration Team
Acknowledgements
