USAF JBAIDS/M1M USAF JBAIDS/M1M Proficiency Test ProgramProficiency Test Program
Ketan Patel, Ph.D.Ketan Patel, Ph.D.
SAFMLSSAFMLS
Feburary 11, 2008Feburary 11, 2008
Points of Interest for Points of Interest for USAF PT Program FY08USAF PT Program FY08
JBAIDS PT Program DetailsJBAIDS PT Program DetailsSubmitting Results for JBAIDS PTSubmitting Results for JBAIDS PTJBAIDS PT FailuresJBAIDS PT FailuresM1M Proficiency Test M1M Proficiency Test AFIP supportAFIP support
JBAIDS PT: Program DetailsJBAIDS PT: Program Details Approximately 53 Sites Approximately 53 Sites Testing FormatTesting Format
2 Unknowns (multiple simulated matrices)2 Unknowns (multiple simulated matrices) 4 Organisms (B.a (2 targets), F.t (1), Y.p (2), 4 Organisms (B.a (2 targets), F.t (1), Y.p (2),
Orthopox(1))Orthopox(1)) 4 Runs per sample (PTC, NTC, Unk1, Unk2; pos, 4 Runs per sample (PTC, NTC, Unk1, Unk2; pos,
neg, and unks ran in dup)neg, and unks ran in dup) 4 Extraction Procedures4 Extraction Procedures
QFlow QFlow ScoopScoop VibeVibe SwipeSwipe
JBAIDS PT: Program DetailsJBAIDS PT: Program Details
Three PT events per fiscal yearThree PT events per fiscal yearSupplies Supplies
Unknown samples (2) Unknown samples (2) DNA or heat-killed bacteria DNA or heat-killed bacteria
Variety of matricesVariety of matrices Sterility reports kept on file at AFIPSterility reports kept on file at AFIP
Provided by AFIPProvided by AFIP Shipped via FEDEXShipped via FEDEX Notification of receipt of samples required from HLDsNotification of receipt of samples required from HLDs
JBAIDS PT: Program DetailsJBAIDS PT: Program Details
ReagentsReagentsExtraction Kits (Extraction Kits (QQFLOW, Scoop, VIBE, and FLOW, Scoop, VIBE, and
Swipe)Swipe) Provided by individual sites Provided by individual sites
JBAIDS Assay TargetsJBAIDS Assay Targets Lyophilized reagents require reconstitutionLyophilized reagents require reconstitution
Reconstitution water provided in assay kitReconstitution water provided in assay kit Reconstitution buffer provided in assay kitReconstitution buffer provided in assay kit
Provided by individual sites Provided by individual sites
JBAIDS PT: Program DetailsJBAIDS PT: Program Details
RT-PCR Using JBAIDS PlatformRT-PCR Using JBAIDS PlatformUnknown sample QC done by AFIPUnknown sample QC done by AFIP
Pre-shipment QCPre-shipment QCPost-shipment QCPost-shipment QC
Results submitted to PT CoordinatorResults submitted to PT CoordinatorDue 10 business days from the time of receipt Due 10 business days from the time of receipt
of PT samples by HLDof PT samples by HLD
Changes With The JBAIDS PT & Changes With The JBAIDS PT & InstrumentationInstrumentation
Sites provide their own reagents for PTSites provide their own reagents for PTJBAIDS software differs from RAPID JBAIDS software differs from RAPID
User-friendlyUser-friendlySpecific positioning of samplesSpecific positioning of samplesAutomatically makes result callsAutomatically makes result callsNotifies user of failed QCsNotifies user of failed QCsProduces a “combined call” based on QC Produces a “combined call” based on QC
results and individual sample resultsresults and individual sample results
Changes With The JBAIDS PT & Changes With The JBAIDS PT & Instrumentation (cont.)Instrumentation (cont.)
Multiple extraction protocols for different Multiple extraction protocols for different matricesmatricesQFlow (replaced Flow in Fall 07)QFlow (replaced Flow in Fall 07)ScoopScoopSwipeSwipeVIBE VIBE
QFlow Sample ExtractionQFlow Sample Extraction
MatricesMatricesWhole blood (DNA)Whole blood (DNA)Food (DNA)Food (DNA)
WaterWaterTunaTunaGround BeefGround BeefSaladSalad
Gastric Wash (DNA)Gastric Wash (DNA)Water (DNA)Water (DNA)Air into PBS (DNA/RNA)Air into PBS (DNA/RNA)
Scoop & Vibe ExtractionsScoop & Vibe Extractions
Matrices for Scoop ExtractionMatrices for Scoop ExtractionSoil (DNA)Soil (DNA)Stool (DNA)Stool (DNA)
Matrices for Vibe ExtractionMatrices for Vibe ExtractionWhole Blood (DNA)Whole Blood (DNA)Nasal Swab (RNA)Nasal Swab (RNA)Sputum (DNA/RNA)Sputum (DNA/RNA)
Swipe Sample ExtractionSwipe Sample Extraction
MatricesMatricesNasal/Pus SwabNasal/Pus SwabSurface SwabSurface SwabPowder Powder Culture SamplesCulture Samples
Phases of JBAIDS ExtractionsPhases of JBAIDS Extractions
Reconstitution of samplesReconstitution of samples Sample lysisSample lysis
Bead BeatingBead Beating
DNA/RNA Bound to filterDNA/RNA Bound to filter Buffer 1Buffer 1
DNA/RNA Washed of inhibitorsDNA/RNA Washed of inhibitors Buffer 2Buffer 2
DNA/RNA Eluted from the filterDNA/RNA Eluted from the filter Buffer 3Buffer 3
Submitting ResultsSubmitting Results
Fax Result Forms to AFIPFax Result Forms to AFIPFax Conditions & Progress Logs to AFIPFax Conditions & Progress Logs to AFIPFax JBAIDS report to AFIPFax JBAIDS report to AFIPForward JBAIDS run files (.ixo) to AFIPForward JBAIDS run files (.ixo) to AFIPEvaluation of PTEvaluation of PT
Grades will be determined based on:Grades will be determined based on:JBAIDS reportJBAIDS reportResult form responses Result form responses
>>80% constitute passing grade for PT80% constitute passing grade for PT
FailuresFailures
JBAIDS Failure policyJBAIDS Failure policy (Insufficient) Less than 80% in a PT period (Insufficient) Less than 80% in a PT period
requires:requires:Investigation of failureInvestigation of failurePerform any appropriate action to correct problemPerform any appropriate action to correct problemComplete Exception response formComplete Exception response formReturn form to AFIP PT CoordinatorReturn form to AFIP PT Coordinator
(Poor) Less than 80% in two out of three (Poor) Less than 80% in two out of three consecutive PT periods requires:consecutive PT periods requires:The discontinuation of testing until MAJCOM The discontinuation of testing until MAJCOM
approves resumptionapproves resumptionInvestigation of failureInvestigation of failure
Failure (cont.)Failure (cont.)
Perform any appropriate action to correct problemPerform any appropriate action to correct problemComplete Exception response formComplete Exception response formForward completed Exception report including Forward completed Exception report including
investigation results, corrective action proposals, investigation results, corrective action proposals, etc. to MAJCOM for review and signatureetc. to MAJCOM for review and signature
MAJCOM will decide if appropriate action has MAJCOM will decide if appropriate action has been taken and if testing should resumebeen taken and if testing should resume
Forward forms to AFIP PT Coordinator after Forward forms to AFIP PT Coordinator after MAJCOM approvalMAJCOM approval
Within three days of resumption of testing forward Within three days of resumption of testing forward Exception response form to the CCLM officeException response form to the CCLM office
Failures (cont.)Failures (cont.)
(Unsatisfactory) Less than 80% in three (Unsatisfactory) Less than 80% in three out of four consecutive PT periods out of four consecutive PT periods requires:requires:Complete the steps for poor performance Complete the steps for poor performance
listed previously in addition to listed previously in addition to The Director of the CCLM will review investigations The Director of the CCLM will review investigations
and other corrective action documentationand other corrective action documentationThe decision to resume testing will be made by the The decision to resume testing will be made by the
Director of the CCLMDirector of the CCLM
Current Status of JBAIDS PT ‘08Current Status of JBAIDS PT ‘08
PT events prior to ’08 were educational PT events prior to ’08 were educational challangeschallanges
JBAIDS PT ‘08 #1 JBAIDS PT ‘08 #1 Shipped on January 4, 2008Shipped on January 4, 2008Awaiting final reportAwaiting final report
Common errors from Past PTsCommon errors from Past PTs
Common errors Common errors Failure to follow OIFailure to follow OIFailure to detect DNA (technical)Failure to detect DNA (technical)Blanks left on result form (clerical)Blanks left on result form (clerical)
Common Errors for JBAIDS PTCommon Errors for JBAIDS PT
Failure to follow SOPFailure to follow SOPRunning Target 2 with Target 1 Running Target 2 with Target 1 Target 2 should be ran only when Target 1 is Target 2 should be ran only when Target 1 is
positivepositiveUsing wrong extraction kitUsing wrong extraction kitTranscription errorsTranscription errors
Switching answersSwitching answersSkipping answers (leaving results blank)Skipping answers (leaving results blank)
Current Current
Changes for fall of 2007Changes for fall of 2007 Introduction of inhibition controls for each Introduction of inhibition controls for each
assayassay Introduction of extraction controls for each Introduction of extraction controls for each
extractionextractionSites will stock the following assays onlySites will stock the following assays only
Ba target #1 and #2Ba target #1 and #2Yp target #1 and #2Yp target #1 and #2F. tularensisF. tularensisOrthopoxOrthopoxSmallpox Smallpox
M1M Proficiency TestingM1M Proficiency Testing
Approximately 27 participantsApproximately 27 participantsTesting FormatTesting Format
One unknownOne unknownThree target toxinsThree target toxins
M1M Test Menu for HLDM1M Test Menu for HLDRicinRicinBot ABot ASEB (Staphylococcus enterotoxin B)SEB (Staphylococcus enterotoxin B)
M1M Proficiency TestM1M Proficiency Test(Cont.)(Cont.)
SuppliesSupplies UnknownsUnknowns
Non-infectious, lyophilized materialNon-infectious, lyophilized material Provided by AFIPProvided by AFIP
Reagents and Instrument suppliesReagents and Instrument supplies Provided by individual sitesProvided by individual sites
Frequency and Delivery MethodsFrequency and Delivery Methods Three times per fiscal yearThree times per fiscal year M1M PT will be delivered with JBAIDS PTM1M PT will be delivered with JBAIDS PT
M1M PT will be scored independently of JBAIDS PTM1M PT will be scored independently of JBAIDS PT
M1M Proficiency Test M1M Proficiency Test (Cont.)(Cont.)
Submitting resultsSubmitting resultsFax result form to AFIPFax result form to AFIPFax conditions and progress logs to AFIPFax conditions and progress logs to AFIP
GradingGradingPass/Fail ScoringPass/Fail Scoring
Based on number of unknowns and assaysBased on number of unknowns and assaysWith only 3 possible answers, one incorrect result With only 3 possible answers, one incorrect result
causes the percentage to fall below 80%causes the percentage to fall below 80%
Common Errors with M1M PTsCommon Errors with M1M PTs
Problems that occurredProblems that occurredLack of package inserts with controlsLack of package inserts with controlsLack of OILack of OILack of “key” to coded reagentsLack of “key” to coded reagentsError in reconstitution directions for PT Error in reconstitution directions for PT
materialmaterial
AFIP SupportAFIP Support
AFIP PT CoordinatorAFIP PT Coordinator Notification of shipment of PT thru emailNotification of shipment of PT thru email Technical supportTechnical support
Via emailVia email Via phoneVia phone
Tracking information for shipments will be made Tracking information for shipments will be made available upon requestavailable upon request
Distribution of results to individual sites and MAJCOM Distribution of results to individual sites and MAJCOM consultants via emailconsultants via email
Limited number of replacement PT samples will be Limited number of replacement PT samples will be available upon requestavailable upon request
AFIP Proficiency Testing TeamAFIP Proficiency Testing Team
Dr. Ketan PatelMolecular Chief
Ms. Ellen LaMorenaAF PT Coordinator
Dr. S. FrancesconiMicrobiology Chief
Major Patrick KennedyScience Coordinator
Dr. Robert CrawfordDivision Chief
AFIP
AFIP Contact InformationAFIP Contact Information Dr. Robert CrawfordDr. Robert Crawford
(202)782-1825(202)782-1825 [email protected]@afip.osd.mil
Dr. Ketan PatelDr. Ketan Patel (202) 782-2847(202) 782-2847 [email protected]@afip.osd.mil
Major Patrick KennedyMajor Patrick Kennedy (202) 782-1863(202) 782-1863 [email protected]@afip.osd.mil
Dr. Stephen FrancesconiDr. Stephen Francesconi (202)782-1834(202)782-1834 [email protected]@afip.osd.mil
Ms. Ellen LaMorenaMs. Ellen LaMorena (202) 782-1861(202) 782-1861 [email protected]@afip.osd.mil