Mira Mandelbaum-Livnat, Efrat Barbiro-Michaely and Avraham Mayevsky
National Institute of Allergy and Infectious Diseases ... · 11/6/2002 · IQA Project Manager...
Transcript of National Institute of Allergy and Infectious Diseases ... · 11/6/2002 · IQA Project Manager...
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National Institute of Allergy and Infectious Diseases Division of
AIDS - Immunology Quality Assessment ProgramAt the Center for Laboratory Investigation - New Jersey
Medical School - Newark, NJAccredited by CAP, Licensed by NJ Dept. of Health, CLIA Approved
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IQA MISSION
The IQA is a resource designed to help ACTG
Immunologists evaluate and enhance theintegrity and comparability of immunologicallaboratory determinations performed onpatients enrolled in multi-site HIV/AIDSinvestigations (therapeutic, vaccine,prevention, etc.).
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IQA Program Organization
FSTRFAMHERST, NY
CENTER for LABORATORYINVESTIGATION
NEWARK, NJ
IQA PROGRAM
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IQA OPERATIONS STRUCTURE
SDAC
FSTRFData Management
CLI UnitsLaboratory Support
IQA Project ManagerRaul Louzao
IQA PIThomas N. Denny
NIAIDPROJECT OFFICER
Daniella Livnat
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CLI Flow Cytometry Unit• Multi Color Immunophenotyping• Sorting• Intracellular Cytokines• Flow Cytometric Evaluation Studies
(e.g.Rainbow Bead Study)
• Tetramer• Evaluation of Alternative messurements of CD4
(Facscount, Guava, Dynabeads, Cytospheres)
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CLI Functional Immunology Unit
• Lymphocyte Proliferative Assays (LPA)– HIV Antigen Evaluation
• HIV p24 (Protein Science, Austral)• HIV gp120 (Protein Science, Austral)• HIV-1 MN (NCI)
• LPA Instrument Evaluation• ELISPOT• Cryopreservation
– Viability and Viable Cellular Rcovery
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IQA/ACTG Projects
• CD4/CD8 QA Bi-Monthly Sendout (83 National Laboratories)– Advance Flow Evaluation
• Reviewed Histograms– Troubleshooting Problem Sites
• PICL Cryopreservation Study 3 Color Immunophenotyping (WB and PBMCs)– LPA and Flow Cytometry(Fresh PBMCs and Frozen/Thawed
PBMCs)– Freezing, Thawing and Viability Assessment
• Adult ACTG Cryopreservation Study (38 National Laboratories)
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IQA/ACTG Projects
• LPA CMV QA • LPA HIV Ag QA Study
– HIV PCR RNA/ CD4 Evaluation on 15 Donors– Four Different Ag Concentrations Day 4 and Day 6– Protein Science (HIV p24, HIV gp120)– Austral (HIV p24, HIV gp 120)
• ELISPOT QA Sendouts (7 Pediatric ACTG Laboratories)• PLG (CD4 Gating Strategy)• CD4 International Validation Study (Bali 4/2002 and 4/2003)• Evaluation of Whole Blood Stabilizers• Validation of Simpler CD4 T cell Substets
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Bali Results 6/11/02Received Processing Averagedate/time date/time count cell/ul count cell/ul count cell/ul cell/ul
1 020601 10/6/02, 16.00 pm 11/06/02, 8.30 am 137 152 148 164 1582 020602 10/6/02, 16.00 pm 11/06/02, 8.30 am 535 594 411 457 5263 020603 10/6/02, 16.00 pm 11/06/02, 8.30 am 101 112 99 110 1114 020605 10/6/02, 16.00 pm 11/06/02, 10.15 am 62 69 71 79 745 020606 10/6/02, 16.00 pm 11/06/02, 10.15 am 234 260 227 252 2566 020607 10/6/02, 16.00 pm 11/06/02, 10.15 am 78 87 81 90 887 020609 10/6/02, 16.00 pm 11/06/02, 11.40 am 33 37 38 42 398 020610 10/6/02, 16.00 pm 11/06/02, 11.40 am 122 136 126 140 1389 020611 10/6/02, 16.00 pm 11/06/02, 11.40 am 66 73 70 78 7610 020613 10/6/02, 16.00 pm 11/06/02, 11.40 am 88 98 99 110 10411 020614 10/6/02, 16.00 pm 11/06/02, 12.50 pm 313 348 328 364 35612 020615 10/6/02, 16.00 pm 11/06/02, 12.50 pm 65 72 81 90 8113 020617 10/6/02, 16.00 pm 11/06/02, 12.50 pm 63 70 60 67 6814 020618 10/6/02, 16.00 pm 11/06/02, 12.50 pm 173 192 164 182 18715 020619 10/6/02, 16.00 pm 11/06/02, 14.10 pm 71 79 91 101 9016 020621 10/6/02, 16.00 pm 11/06/02, 14.10 pm 81 90 77 86 8817 020622 10/6/02, 16.00 pm 11/06/02, 14.10 pm 182 202 198 220 21118 020623 10/6/02, 16.00 pm 11/06/02, 14.10 pm 64 71 69 77 7419 020625 10/6/02, 16.00 pm 11/06/02, 15.40 pm 53 59 55 61 6020 020626 10/6/02, 16.00 pm 11/06/02, 15.40 pm 285 317 321 357 33721 020627 10/6/02, 16.00 pm 11/06/02, 15.40 pm 93 103 87 97 10022 020629 10/6/02, 16.00 pm 11/06/02, 15.40 pm 51 57 43 48 5223 020630 10/6/02, 16.00 pm 11/06/02, 15.40 pm 302 336 329 366 35124 020631 10/6/02, 16.00 pm 11/06/02, 15.40 pm 76 84 72 80 82
Control 464 516 409 454 485
Sample IDNo. Count 1 Count 2 Count 3
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IQA Results Over 3 day period
CD4 % Abs # Lymphs Abs# CD4 CD4 % Abs # Lymphs Abs# CD4 CD4 % Abs # Lymphs Abs# CD4 020601 19 624 120 18 609 109 34 542 182
020602 51 1250 632 49 1210 597 54 1990 1081
020603 21 1030 218 21 1070 222 21 968 208
020605 19 607 114 17 881 148 33 837 277
020606 49 1390 680 52 1380 712 52 1500 787
020607 20 1110 221 20 1050 212 22 843 182
020609 19 472 92 19 929 177 37 556 204
020610 51 1580 811 52 1280 662 53 1300 691
020611 21 986 207 22 836 182 22 933 207
020613 20 704 144 15 640 97 35 551 192
020614 49 1270 621 50 1070 531 53 1380 728
020615 20 722 147 22 915 198 22 853 184
020617 20 676 136 15 980 146 33 392 131
020618 51 1270 651 50 1160 585 52 1460 765
020619 20 943 189 20 902 184 21 759 163
020621 18 364 65 16 632 98 31 441 139
020622 50 1580 793 51 1100 561 52 1350 707
020623 21 1060 220 21 944 197 22 880 191
020625 18 938 168 14 883 126 33 410 133
020626 52 1370 714 50 1110 551 54 1333 724
020627 22 981 216 19 1020 199 24 960 231
020629 19 1030 191 14 582 82 31 510 159
020630 49 1450 704 50 1330 671 51 1310 669
020631 22 827 183 21 879 181 22 944 205
IQA Counts
IQA Day 1 6/4/02 IQA Day 3 6/6/02 IQA Day 7 6/10/02Sample ID
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BALI/IQA Comparison
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CD4
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ACTG Statistical Results4/11/2003
Sample Number CD4% CD8% WBC %Lymph Total Lymph CD4 # CD8#
30401 46.95 18.11 3040 0.483 1468 689 44630405 44.7 17.27 2990 0.481 1438 643 43030413 45.89 17.8 3180 0.484 1539 706 48930409 46.24 17.4 3190 0.492 1569 726 501
SD 0.94 0.38 100 0.00 61 35 34Mean 45.95 17.65 3100 0.49 1504 691 467Median 46.07 17.60 3110 0.48 1504 698 468
4/14/03Sample Number CD4% CD8% WBC %Lymph Total Lymph CD4 # CD8#
30401 47.08 17.04 2750 0.434 1194 562 32830409 45.84 16.63 1670 0.693 1157 531 19330413 45.55 18.01 1780 0.765 1362 620 24230405 47.49 17.62 1770 0.778 1377 654 244
SD 0.94 0.61 507 0.16 113 56 56Mean 46.49 17.33 1993 0.67 1272 592 252Median 46.46 17.33 1775 0.73 1278 591 243
4/16/03Sample Number CD4% CD8% WBC %Lymph Total Lymph CD4 # CD8#
30401 45.06 17.84 2640 0.391 1032 465 27330405 44.89 16.94 1550 0.777 1204 541 18730409 43.2 16.58 1770 0.778 1377 595 24430413 43.77 17.49 2670 0.439 1172 513 313
SD 0.89 0.56 582 0.21 142 54 53Mean 44.23 17.21 2158 0.60 1196 528 254Median 44.33 17.22 2205 0.61 1188 527 258
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Bali/IQA 4/2003 Comparison
Bland-Altman NJ/Prodia April 2003
-200-100
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Average of NJ & Prodia results
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Prodia and NJ CD4 results, April 2003
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IQA –International Projects
• CD4 International Validation Study• Evaluation of Whole Blood Stabilizers• Validation of Simpler CD4 T cell Subsets• Designing laboratories for resource poor countries
– Training personnel – Suggesting equipment purchases– Validating laboratory
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InternationalGeorgetown Hospital
Complex• Evaluate locations for laboratory • Recommend equipment• Establish SOP and Job descriptions• Design Laboratory layout• Establish timeline for training • Develop training schedule
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The top floor is the lab space. These pictures are taken from different angles.
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Top 2 pictures represents the mail lab area of the building
Housing for the electrical generator (outside shot and inside shot on the right)
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Laboratory
Court Yard