EXTERNAL QUALITY ASSESSMENT SCHEME

Evaluation Booklet

CARIBBEAN REGION

Performance Evaluation Panel OPS0110 C Coordination: Fundação Pró-Sangue Hemocentro de São Paulo Pan American Health Organization International Consortium for Blood Safety Caribbean Epidemiology Centre

São Paulo, 20 July 2010

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Table of Contents

Participant Laboratories ................................…….......... 3 Panel OPS0110 C target result ……..............….…......... 4 Panel OPS0110 C characterization tests results.……...... 4 Syphilis..........................................................….....…... 4 HIV......................................…....…....….………......... 5 Ant-HTLV-I/II........................….…............….…....... 5 HCV.......................................…......………...…......…. 5 HBsAg.........................................……..........……....… 6 Ant-T. cruzi………………………………………..…. 6 Confirmatory test …….................…..............……....... 7 Overall Results…………….................…..........….......... 8 Parameter …...................................…..................……. 8 Participants …………….……………...………..……. 9 Survey Sample …….....…..................…...............…… 10 Test kits.....….......................................….............…… 11 Supplemental test………………………………………. 13

GLOSSARY

CO = Cut off OD = Optical Density IFI = Indirect Immunofluorescence FNR = False Negative Result FPR = False Positive Result TND = Total of Determinations in Negative Samples TPD = Total of Determinations in Positive Samples

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COUNTRY PARTICIPANTS

Anguilla Princess Alexandra Hospital

Antigua Mount St. Johns Medical Center

Bahamas Rand Memorial Hospital

Bahamas Princess Margaret Hospital

Barbados Queen Elizabeth Hospital

Belize Central Medical Laboratory

Cayman Islands Cayman Islands Health Services, George Town Hospital

Netherlands Antilles Red Cross Blood Bank Foundation

Dominica Medical Laboratory, Princess Margaret Hospital

Grenada Pathology Laboratory, General Hospital

Guyana National Blood Transfusion Service, Public Hospital

Jamaica National Public Health Laboratory

Monserrat Pathology Laboratory

Nevis Alexandra Hospital Laboratory

Saint Lucia Ezra Long Laboratory, Victoria Hospital

St. Kitts Joseph N. France General Hospital

St. Vincent and The Grenadies Pathology Laboratory, Milton Cato Memorial Hospital

Suriname National Blood Bank Suriname Red Cross Society

Turks & Caicos Islands Grand Turk Hospital Laboratory

Trinidad National Blood Transfusion Center One performance evaluation panel (OPS0110C) containing 6 samples with positivity to HIV, HCV, ant-HTLV, Syphilis, ant-T.cruzi and HBsAg were shipped to each participant. The survey samples were undiluted, defibrinated plasma obtained from individual donors and containing 0.05% of Bronidox –L5 as a preservative. Of the 20 participants that were sent the performance evaluation panel, 4 of them (20%) did not return the results.

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Panel OPS0110 C – Target Result

Sample Syphilis HIV ant-HTLV HCV HBsAg ant-T. cruzi 1 X X 2 X X 3 X 4 X X 5 X 6 X X

Characterization tests results of the Performance Evaluation Panel OPS0110 C Tables 1 to 9 show the characterization tests results performed at Fundação Pró-Sangue Hemocentro de São Paulo – Quality Control Department. Optical density and cut-off values of the assays are presented for the 6 samples that comprise the Performance Evaluation Panel OPS0110 C. Table 1 Syphilis

Enzygnost Syphilis Architect Syphilis VDRL Test

39103 77033LP64 0907030290 OD CO CO/OD S CO S/CO Result

1 1.649 1.007 0.611 0.590 1.00 0.590 Negative 2 0.090 1.007 11.189 22.600 1.00 22.600 1/16 3 1.647 1.007 0.611 0.410 1.00 0.410 Negative 4 1.542 1.007 0.653 0.740 1.00 0.740 Negative 5 1.491 1.007 0.675 0.250 1.00 0.250 Negative 6 0.060 1.007 16.783 24.970 1.00 24.970 1/8

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Table 2 HIV

Vironostika HIV UniForm II

Plus O Architect HIV Ag/Ab A60BC 75843HN00 OD CO OD/CO OD CO OD/CO

1 3.000 0.169 17.751 476.240 1.00 476.240 2 0.063 0.169 0.373 0.250 1.00 0.250 3 0.060 0.169 0.355 0.210 1.00 0.210 4 0.066 0.169 0.391 0.250 1.00 0.250 5 0.056 0.169 0.331 0.210 1.00 0.210 6 0.059 0.169 0.349 0.120 1.00 0.120

Table 3 Ant - HTLV

Ortho HTLV-I/HTLV-II

Ab-Capture HTV201 OD CO OD/CO

1 3.000 0.322 9.317 2 0.156 0.322 0.484 3 0.186 0.322 0.578 4 0.168 0.322 0.522 5 1.993 0.322 6.189 6 0.176 0.322 0.547

Table 4 HCV

Monolisa HCV Ag-Ab Ultra Ortho HCV ELISA

9C0535 EXE182 OD CO OD/CO OD CO OD/CO

1 0.313 0.396 0.790 0,234 0,602 0,389 2 2.438 0.396 6.157 3,000 0,602 4,983 3 0.263 0.396 0.664 0,185 0,602 0,307 4 2.290 0.396 5.783 3,000 0,602 4,983 5 0.158 0.396 0.399 0,049 0,602 0,081 6 2.486 0.396 6.278 3,000 0,602 4,983

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Table 5 HBsAg

Hepanostika HBsAg

Ultra

Architect HBsAg

B10CB 77476LF00 OD CO OD/CO OD CO OD/CO

1 0.036 0.061 0.590 0.010 0.05 0.010 2 0.028 0.061 0.459 0.010 0.05 0.010 3 0.046 0.061 0.754 0.010 0.05 0.010 4 2.783 0.061 45.623 4.210 0.05 4.210 5 0.052 0.061 0.852 0.010 0.05 0.010 6 0.029 0.061 0.475 0.010 0.05 0.010

Table 6 Ant - T. cruzi

I.F.I. Elisa cruzi Chagas Test Elisa III Biomerieux 0907101512 0271108 Result OD CO OD/CO OD CO OD/CO

1 Negative 0.154 0.258 0.597 0.080 0.418 0.191 2 Negative 0.051 0.258 0.198 0.104 0.418 0.249 3 1/160 2.034 0.258 7.884 1.424 0.418 3.407 4 Negative 0.190 0.258 0.736 0.137 0.418 0.328 5 Negative 0.063 0.258 0.244 0.101 0.418 0.242 6 Negative 0.063 0.258 0.244 0.104 0.418 0.249

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Confirmatory Tests Table 7 HIV 1+2

Western Blot HIV – Genelabs

lot: AE9021 p17 p24 p31 p39 gp41 p51 p55 p66 gp120 gp160 HIVII Result

1 + + + + + + + + + + - Positive Table 8 HTLV I/II

Western Blot HTLV Genelabs Lot: AK9001 GD21 P19 P24 P26 P28 P32 P36 GP46 P53 RGP46II RGP46I Result

1 + + + - - - - - - + - Positive 5 + +/- + - - - - - - - + Positive

Table 9 Hepatitis C

CHIRON RIBA HCV 3.0 SIA Lot: 98743 C100 (P) C33C C22 (P) NS5 hSOD Result

2 4+ 4+ 3+ 4+ - Positive 4 - 2+ 4+ 4+ - Positive 6 4+ 4+ 4+ 4+ - Positive

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OVERALL RESULTS

Graph 1 shows the number of the tests performed by the 16 participants to each parameter. All 16 respondents performed tests for Syphilis and HBsAg. Fifteen respondents performed test for HIV and HCV, eleven respondents performed test for ant-HTLV and two for ant-T. cruzi. The table 10 shows the false positive and false negative results reported for each parameter. The total of the false negative results reported was 1.8% in 225 determinations performed in positive samples and 5.2% of false positive result in 383 determinations performed in negative samples.

Results reported for each parameter

Table10

Percentage of false negative and false positive results by parameter False Positive % False Negative %

Syphilis 1 1.32 0 0 TND = 76 TPD = 53

HIV 2 2.0 0 0 TND = 100 TPD = 27

Ant-HTLV 0 0 1 2.6 TND = 48 TPD = 38

HCV 12 21.8 0 0 TND = 55 TPD = 75

HBsAg 5 5.3 3 10.7 TND = 94 TPD = 28

Ant-T. cruzi 0 0 0 0 TND = 10 TPD = 4

Total 20 5.2 4 1.8 TND = 383 TPD = 225

TND = Total of determinations performed in negative samples TPD = Total of determinations performed in positive samples

02468

1012141618

Syphilis HIV ant-HTLV HCV HBsAg ant-T. cruzi

Parameter

nº L

abor

ator

ies

Graph 1

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Performance result of the participants The performance qualification “A” was attributed to 9 participants to all tests reported. Five participants had good performance. but reported a few false positive results. Qualification “B1” or “B2” was attributed to them. Qualification “C” was attributed to two participants. Table 11 Participants performance by parameter

Participants HIV HBsAg Syphilis HCV Anti-HTLV ant-T. cruzi 1 A A A A A A 2 A A A A A - 3 A A A A - A 4 A A A A A - 5 A A A A A - 6 A A A A A - 7 A A A A - - 8 A A A A - - 9 A A A - - - 10 B1 A A A - - 11 - A B2 A A - 12 A A A B2 A - 13 A A A A A - 14 A B2 A B2 A - 15 A A A A C - 16 B2 C A A A -

A = 100% of concordance B1 = ≤ 5% of the total of determinations B2 = > 5% of the total of determinations C = False negative result -- = Not performed

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Analysis by Sample Graph 2 show the percentage of the false positive results reported to each sample and the Table 12 shows the rate of false positive results of each sample by parameter. False Positive results (%) reported by sample

Table 12 Rate of false positive results of each sample by parameter.

TND = Total of determinations performed in negative samples TPD = Total of determinations performed in positive samples F.P. = False positive results F.N. = False negative results

6,90

0,00

7,95

0,00

8,86

3,77

0,00 0,00 0,00

5,665,26

0,000123456789

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1 2 3 4 5 6

Sample

%

% FP% FN

Graph 2

Sample F.P. F.N. F.P. F.N. F.P. F.N. F.P. F.N. F.P. F.N. F.P. F.N. TPD TND1 6.9 46 582 52 523 2.3 4.6 1.1 4 884 5.7 53 535 2.5 1.3 5.1 5.3 19 796 3.8 51 53

Total 0.5 1.3 1.3 3.1 0.4 0.3 225 383

ParametersHIV HBsAg HCV Ant-HTLV Syphilis T. cruzi

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Analysis by KIT Tables 13 to 18 show the number of the false positive results reported for each kit used by the participants. Table 13 Syphilis

KITS FPR % TND FNR % TPD

Ampcor RPR Card Test 0 0 16 0 0 10 VDRL Antigen Test for Syphilis Screening 0 0 8 0 0 6 ICE Syphilis 0 0 8 0 0 6 ATI Syphilis RPR 0 0 4 0 0 4 RPR NOSTICON II 0 0 4 0 0 4 SERODIA - TP-PA 0 0 4 0 0 4 VDRL Antigen for Slide Test 0 0 4 0 0 4 Becton Dickson VDRL antigen 0 0 4 0 0 3 Determine Rapid Syphilis TP Asay 0 0 4 0 0 2 Macrovue RPR Card 0 0 4 0 0 2 RPR Biokit 0 0 4 0 0 2 Stanbio RPR Quich Test 1 25 4 0 0 2 Trepanostika TP Recombinante 0 0 4 0 0 2 VDRL Cardiolipin antigen - Difco 0 0 4 0 0 2

TOTAL 1 1.3 76 0 0 53 Table 14 HIV

KITS FPR % TND FNR % TPD

Vironostika HIV Uni-Form II Ag/Ab 0 0 25 0 0 8 Axsym HIV Ag/Ab COMBO 0 0 15 0 0 4 HIV Ag/Ab Architect i2000 SR 0 0 15 0 0 4 Determine Rapid HIV 1/2 2 13.3 15 0 0 3 ACCESS HIV 1/2 0 0 5 0 0 1 Axsym HIV-1/HIV-2 0 0 5 0 0 1 Elecsys HIV Combi 0 0 5 0 0 1 Genscreen Ultra HIV Ag/Ab 0 0 5 0 0 2 HIV Combinado Ag/Ab - Murex 0 0 5 0 0 2 Uni-Gold HIV Rapid Test 0 0 5 0 0 1

TOTAL 2 2.0 100 0 0 27 Table 15 Ant - HTLV

KITS FPR % TND FNR % TPD

Murex HTLV I+II 0 0 28 0 0 22 DiaPro HTLV I/II Ab 0 0 8 1 16.7 6 HTLV I/II Architect i 2000 SR 0 0 4 0 0 4 HTLV I/II rec. Biokit 0 0 4 0 0 4 Orgenics ImmunoComb II 0 0 4 0 0 2

TOTAL 0 0 48 1 2.6 38

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Table 16 HCV

KITS FPR % TND FNR % TPD

Axsym HCV V.3 12 100 12 0 0 12 Murex anti-HCV (versão 4.0) 0 0 10 0 0 15 anti-HCV Architect i2000 SR 0 0 9 0 0 12 Hepanostika HCV Ultra 0 0 9 0 0 18 Orgenics Rapid Signal 0 0 6 0 0 6 Elecsys HCV 0 0 3 0 0 6 VITROS ECI para anti-hcv 0 0 3 0 0 3 AKUDEX HCV 0 0 3 0 0 3

TOTAL 12 21.8 55 0 0 75

Table 17 HBsAg

KITS FPR % TND FNR % TPD

Murex HBsAg Versão 3 0 0 20 0 0 6 Hepanostika HBsAg Ultra 1 5.9 17 0 0 5 Axsym HBsAg (v2) 4 23.5 17 0 0 5 Access HBsAg 0 0 5 0 0 1 AKUCHECK HBsAg 0 0 5 1 100 1 Cobas Core HBsAg II 0 0 5 0 0 1 Determine Rapid HBsAg 0 0 5 2 100 2 Elecsys HBsAg 0 0 5 0 0 2 HBsAg Architect i2000 SR 0 0 5 0 0 2 MONOLISA AgHBs Ultra 0 0 5 0 0 2 VITROS ECI para HBsAg 0 0 5 0 0 1

TOTAL 5 5.32 94 3 10.71 28 Table 18 Ant - T. cruzi

KITS FPR % TND FNR % TPD

Chagatek Elisa 0 0 5 0 0 1 Chagatest Elisa Recombinante v.3.0 0 0 5 0 0 3

TOTAL 0 0 10 0 0 4

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Supplemental tests Table 19 shows the confirmatory tests performed by the participants. Table 19 Supplement tests performed by the participants by parameter

Parameter Kit Nº of Participants

Second different ELISA Test Kit 1

HIV Second different rapid test 1

Rapid test 1

INNO-LIATM HIV I/II Score 1

HBsAg Murex HBsAg Confirmatory v.3 1 HCV INNO-LIATM HCV Score 1

Ant-HTLV I/II HTLV Blot 2.1 1

Serodia TP-PA 1

Syphilis Becton Dickson VDRL antigen 1

ATI Syphilis RPR test 2

Responsable Dra. Márcia Otani Serology - Quality Control Department Fundação Pró-Sangue Hemocentro de São Paulo. Av. Dr. Enéas de Carvalho Aguiar 155 – Cerqueira Cesar – São Paulo. CEP: 05403-000 Tel: 55 11 3061 5544 ext. 353 Fax: 55 11 3088 8317 e-mail: otanimarcia@prosangue.sp.gov.br or otanimarcia@uol.com.br