USAF JBAIDS/M1M Proficiency Test Program

Presented By

Thomas Shaak, Maj, USAF, BSCDeputy Chief, Division of Wound Biology and Translational

ResearchArmed Forces Institute of Pathology

Washington, DC

Purpose of AFPT Program

• AFPT program for the JBAIDS/M1M is a corner stone of HSMR-LBDT capability.

• The PT challenges ensure that in the unlikely event of Biowarfare (BW)/Toxin attack we can accurately indentify the presence of BW agents.

AFPT Program

• 55 Air Force Sites (Bagram and Moody) • Biological and Toxin testing Platforms• JBAIDS proficiency testing• M1M proficiency testing • AFIP support

JBAIDS/M1M

• JBAIDS-uses principles of real-time PCR for presumptive identification of biological warfare agents

• M1M-uses principles of immuno-based ECL for detection of pathogens and toxins in biological, environmental and food samples.

• Both systems allow for rapid ID and diagnostic confirmation of biological pathogens

JBAIDS PT: Program Details• 3 PT events per fiscal year• Real-time PCR platform (JBAIDS)• Testing Format

2 (or more) Unknowns (multiple simulated matrices)4 Organisms (B.a (2), F.t (1), Y.p (2), Orthopox(1))4 Runs per sample (PTC, NTC, Unk1, Unk2; pos, neg, and

unknowns run in duplicate)• Unknown sample QC done by AFIP

8% of samples made, tested for QC• Results submitted to PT Project Manager

Due 10 business days from the time of receipt of PT samples by the HSMR-LBDT sites

JBAIDS PT: Program Details

• Supplies Unknown samples (2+) • Inactivated antigen

Critical Reagent ProgramVariety of matrices

• Provided by AFIPShipped via FEDEXNotification of receipt of samples required from HSMR-LBDT

JBAIDS PT: Program Details

• ReagentsJBAIDS Assay Targets

• Lyophilized reagents require reconstitutionReconstitution water provided in assay kitReconstitution buffer provided in assay kit

• Provided by AFIP (including inhibition and extraction controls) Prior provided by individual sites

Extraction Kits for different matrices (QFLOW, Scoop, VIBE, and Swipe)• Provided by individual sites

QFlow Sample Extraction

• MatricesWhole blood (DNA)Food (DNA)• Water• Tuna• Ground Beef• Salad

Gastric Wash (DNA)Water (DNA)Air into PBS (DNA/RNA)

Scoop & Vibe Extractions

• Matrices for Scoop ExtractionSoilStoolSand (new)

• Matrices for Vibe ExtractionWhole BloodNasal SwabSputum

Swipe Sample Extraction

• MatricesNasal/Pus SwabSurface SwabPowder Culture Samples

JBAIDS Results and Scoring• Submitting Results

Fax to AFIP:• Result Forms• Conditions & Progress Logs• JBAIDS report

Forward JBAIDS run files or paper reports (.ixo) to AFIP

• ScoringGrades will be determined based on:

• JBAIDS report• Result form responses

>80% constitute passing grade for PT

M1M PT: Program Details

• Approximately 47 sites (Bagram)• Three PT events per fiscal year

M1M PT will be delivered with JBAIDS PT• M1M PT will be scored independently of JBAIDS PT

• Immunoassay platform testing formatOne unknownThree target toxins

• Ricin• Bot A• SEB (Staphylococcus enterotoxin B)

M1M PT: Program Details

• SuppliesUnknowns

• Non-infectious, lyophilized material• Provided by AFIP

Reagents and Instrument supplies• Provided by individual sites

M1M Results and Scoring

• Submitting results Fax to AFIP

• Result form• Conditions and progress logs

Forward to AFIP M1M run files or paper reports

• GradingPass/Fail Scoring

• Based on number of unknowns and assays• With only 3 possible answers, one incorrect result causes the

percentage to fall below 80%

AFIP Support• USAF PT Project Manager (Ellen LaMorena)

Notification of shipment of PT thru emailTechnical support

• Via email• Via phone

Tracking information for shipments will be made available upon request

Distribution of results to individual sites and MAJCOM consultants via email

Limited number of replacement PT samples will be available upon request

AFIP will provide additional testing samples for additional training

AFIP Support• New• Site Visits

Remediation or retraining Site request required (notification of Sq/cc)E-mail AFIP support to initiateNotify Chain of Command of the dateAFIP provides onsite training, evaluation, report

and outbrief to site command

Failures (Insufficient) Less than 80% in a PT period requires:

• Investigation of failure• Perform any appropriate action to correct problem• Complete Exception response form• Return form to AFPT Project Manager

(Poor) Less than 80% in two out of three consecutive PT periods requires:• The discontinuation of testing until MAJCOM approves resumption• Investigation of failure

Failure (cont.)• Perform any appropriate action to correct problem• Complete Exception response form• Forward completed Exception report including

investigation results, corrective action proposals, etc. to MAJCOM for review and signature• MAJCOM will decide if appropriate action has been

taken and if testing should resume• Forward forms to AFIP PT Coordinator after MAJCOM

approval• Within three days of resumption of testing forward

Exception response form to the CCLM office

Failures (cont.)

• (Unsatisfactory) Less than 80% in three out of four consecutive PT periods requires:Complete the steps for poor performance listed previously

in addition to • The Director of the CCLM will review investigations and other

corrective action documentation• The decision to resume testing will be made by the Director of the

CCLM

Common Errors for PT

• Failure to follow SOP• Using wrong extraction kit (JBAIDS)• False positives/negatives– In a real scenario-a false positive could create an

international incident and a false negative could lead to unnecessary casualties

• Transcription errors– Switching answers– Skipping answers (leaving results blank)

Results from AFPT-July 2009

• JBAIDS PT:54 out of 54 HSMR-LBDT sites participated 93% passing (4 out of 54 failed)• 4 failed sites: 4 false negative responses• 2 sites were second failures• All sites repeated and did not pass

Results from AFPT July 2009

• M1M PT:46 HLD sites currently have M1M platform36 out of 46 HLD sites participated

• 100% passing (36 out of 36)• 10 non-participating sites--due to lack of reagents or instrument

maintenance

Changes to AFPT Scoring

• No Changes

Future Direction of PT

• More challenging• Inhibitors

Questions???