NORIP - Haematology Preliminary results from the Nordic Reference Interval Project
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1 NORIP - Haematology Preliminary results from the Nordic Reference Interval Project Arne Mårtensson Clinical Biochemist, EQUALIS, Sweden
2 Working group NORIP Haematology: Eeva-Riitta Savolainen, Oulu/Uleåborg, FI Veli Kairisto, Turku/Åbo, FI Niels Jørgen Christensen, Aarhus/Århus, DK Leifur Franzson, Reykjavik, IS Vigfus Thorsteinsson, Akureyri, IS Sverre Sandberg, Bergen, NO Marthe Wedø Aune, Trondheim, NO Birgitta Swolin, Gothenburg/Göteborg, SE Kurt Karlsson, Umeå, SE Gunnar Nordin, Uppsala, SE Arne Mårtensson, Uppsala, SE
3 NORIP - Haematology 60 Nordic laboratories Denmark: 8 Finland: 26 Iceland: 1 Norway: 1 Sweden: 24 Ca reference persons Ca results Finland Sample collection day Local measurement Day 2 Local and central measurement Other countries Sample collection day: Local measurement
4 Country Number of results Denmark 230 Finland 854 Iceland 78 Norway 31 Sweden 677 Most influence on the common results from Finland and Sweden.
5 Inclusion criteria for reference persons The reference individual should be feeling subjectively well have reached the age of 18 not be pregnant or breast-feeding not have been an in-patient in a hospital nor have been subjectively dangerously ill during the last month not have consumed more than 2 measures of alcohol (24 g) in the last 24 hours not have given blood as a donor in the last five months not have taken prescribed drugs other than the P-pill or estrogens during the last two weeks not have smoked in the last hour prior to blood sampling By courtesy from Pål Rustad
6 Data handling 1. Collecting all results and reference person data into one common database. 2. Correcting results for sample dilution. 3. Excluding outliers. 4. Test for partitioning. 5. Nonparametric calculation of reference intervals robust, not influenced by small numbers of outliers. 6. Checking dependence on country, instrument etc.
7 Correcting results for sample dilution.
8 Influence of the type EDTA-tube on the mean B-Hb? Medelv B-Hemoglobin (g/l) ,000 1,012 1,024 1,038 Spädfaktor ± 95% Confidence interval
9 Component For which components are the results corrected with the dilution factor? Dilution Factor used (3-part) B-Granulocyter Yes (3-part) B-Lymfocyter Yes (3-part) B-Monocyter/MID Yes (5-part) B-Basofila granulocyter Yes (5-part) B-Eosinofila granulocyter Yes (5-part) B-LUC Yes (5-part) B-Lymfocyter Yes (5-part) B-Monocyter Yes (5-part) B-Neutrofila granulocyter Yes B-MPV No B-PDW No B-RDW No B-RDW fl No B-Retikulocyter Yes B-HDW? B-Leukocyter Yes B-Erytrocyter Yes B-Hemoglobin Yes B-Hematocrit Yes B-MCV No B-MCH No B-MCHC No B-Trombocyter Yes
10 Exclusion rules for NORIP Haematology: Single results have been excluded if they deviated more than +/- 5 SD from component mean. Exclusion according to this rule has been repeated until no result was excluded (twice needed). No results has been excluded for other reasons.
11 Results deviating >5 SD, first round Component A_ID Lno COUNTRY Result Deviation SEX (5-part) B-Basofila granulocyter DENMARK 0,2048 5, F (5-part) B-Eosinofila granulocyter DENMARK 13,312 27,32669 M (5-part) B-LUC DENMARK 8,192 6, F (5-part) B-Lymfocyter SWEDEN 7,9948 5, M (5-part) B-Lymfocyter DENMARK 23,4 19,06769 F (5-part) B-Lymfocyter SWEDEN 10,8544 7, M (5-part) B-Lymfocyter SWEDEN 13,494 10,22188 F (5-part) B-Monocyter SWEDEN 1,4168 6, M (5-part) B-Neutrofila granulocyter SWEDEN 9, , F (5-part) B-Neutrofila granulocyter SWEDEN 9,6 5, F B-Hematocrit FINLAND ,45325 F B-Hemoglobin DENMARK 71-5,92328 F B-Leukocyter SWEDEN 15,9744 6, M B-Leukocyter SWEDEN 16,1928 6, F B-Leukocyter DENMARK 27,7 14,13079 F B-Leukocyter FINLAND 16,728 7, M B-MCH SWEDEN 22-5,17498 M B-MCH DENMARK 15,5-9,20839 F B-MCHC DENMARK 269-6,63061 F B-MCHC SWEDEN 240-9,46706 M B-MCV DENMARK 57,6-7,88127 F B-RDW DENMARK 19,5 6, F B-RDW DENMARK 19,7 6, F B-Retikulocyter SWEDEN 302,7968 8, M B-Trombocyter SWEDEN 580,608 5, M
12 Results deviating >5 SD, second round Component Lno PNO Cno COUNTRY Result SEX SD SDII (5-part) B-Eosinofila granulocyter SWEDEN 0,74736 F 1, , (5-part) B-Neutrofila granulocyter ICELAND 8,855 F 4, , (5-part) B-LUC ICELAND 6,3 F 4, ,065325
13 Test for partitioning
14 Percentage (p) of subpopulation results outside the common reference limits determines if separate reference intervals for the subpopulations are recommended: If the big p <3,2 % and the small p 1,8 % Common reference interval is recommended = No partitioning. If the small p 0,9 % but <1,8 % or the big p 3,2 % but <4,1% Separate reference interval is possibly justified = No partitioning is proposed in NORIP Haematology. If the small p <0,9 % or the big 4,1 % Separate reference interval is recommended = Partitioning.
15 By courtesy from Ari Lahti
16 By courtesy from Ari Lahti Proportion (or distance criteria)
17 By courtesy from Ari Lahti
18 By courtesy from Ari Lahti
19 Nonparametric calculation of reference intervals
20 Nonparametric determination of reference limits. Example 1: B-Hb, women, lower limit Limit value at: 2,5 % of totally 960 results = 24,0 Order no Analyte Result AGE SEX 1 B-Hemoglobin 93,0 23 F 2 B-Hemoglobin 102,0 36 F 3 B-Hemoglobin 106,1 53 F 4 B-Hemoglobin 111,2 19 F 5 B-Hemoglobin 113,2 51 F 6 B-Hemoglobin 113,2 51 F 7 B-Hemoglobin 113,3 44 F 8 B-Hemoglobin 114,3 43 F 9 B-Hemoglobin 114,3 28 F 10 B-Hemoglobin 115,3 70 F 11 B-Hemoglobin 115,4 18 F 12 B-Hemoglobin 115,7 44 F 13 B-Hemoglobin 115,7 23 F 14 B-Hemoglobin 115,7 23 F 15 B-Hemoglobin 115,9 50 F 16 B-Hemoglobin 116,0 41 F 17 B-Hemoglobin 116,3 78 F 18 B-Hemoglobin 116,3 46 F 19 B-Hemoglobin 116,3 73 F 20 B-Hemoglobin 116,8 25 F 21 B-Hemoglobin 117,3 62 F 22 B-Hemoglobin 117,3 80 F 23 B-Hemoglobin 117,4 77 F 24 B-Hemoglobin 117,4 24 F 25 B-Hemoglobin 117,8 22 F 26 B-Hemoglobin 117,8 27 F 27 B-Hemoglobin 118,0 79 F 28 B-Hemoglobin 118,3 45 F 29 B-Hemoglobin 118,3 36 F 30 B-Hemoglobin 118,3 21 F
21 Non-parametric determination of reference limits. Example 2: B-Hb, women,high limit Limit value at: 2,5 % of 960 results = 24,0 results Order no Analyte Result 30 B-Hemoglobin 152,0 29 B-Hemoglobin 152,0 28 B-Hemoglobin 152,0 27 B-Hemoglobin 152,6 26 B-Hemoglobin 152,8 25 B-Hemoglobin 153,0 24 B-Hemoglobin 153,0 23 B-Hemoglobin 153,0 22 B-Hemoglobin 153,0 21 B-Hemoglobin 153,6 20 B-Hemoglobin 154,8 19 B-Hemoglobin 155,0 18 B-Hemoglobin 155,0 17 B-Hemoglobin 155,6 16 B-Hemoglobin 155,7 15 B-Hemoglobin 155,7 14 B-Hemoglobin 155,7 13 B-Hemoglobin 156,1 12 B-Hemoglobin 157,1 11 B-Hemoglobin 158,1 10 B-Hemoglobin 159,1 9 B-Hemoglobin 159,1 8 B-Hemoglobin 159,1 7 B-Hemoglobin 159,7 6 B-Hemoglobin 159,9 5 B-Hemoglobin 160,8 4 B-Hemoglobin 161,2 3 B-Hemoglobin 164,2 2 B-Hemoglobin 165,2 1 B-Hemoglobin 166,0
22 Bias between haematology instruments, when comparing all instrument groups with more than 100 results?
23 Code Instrument models in the group Short name -1 Unknown 11 Coulter M2, M4, CBC 5, M Coulter S Coulter SPlus II-VI, STKR, STKS, MaxM, Onyx, GenS Coulter big 14 Coulter T540, T660, T850, TC10, Js, MD8, MD18, MDII 18, AcT 20 Cell-Dyn 300, 400, 500, 610, 700, 800, 900, Cell-Dyn 1300, 1400, 1500, 1600, 1700, Cell-Dyn CS 3000, 3500 Cell-Dyn Cell-Dyn Cell-Dyn 4000 Cell-Dyn ABX Pentra 30 Sysmex CC130, 150, 170, 180, Sysmex F 300, 500, 520, 800, Sysmex K 800, K-1000, K-4500, M-2000, KX-21 Sysmex small 33 Sysmex NE 1500, NE 8000, SE 9000, SF 3000, XE 2100 Sysmex big 40 Bayer H-1, H-1 Junior, H-2, H-3 Bayer H 41 Bayer Advia Bayer Advia 50 Cobas Micros, Minos ST, STE, STX 51 Cobas Argos, Helios, Vega 60 Medonic Ca 460, 480, 580A, Celltac MEK 6108K, 8118K 68 Serono 70 QBC II Plus 80 Swelab 85 Contraves Digicell 88 Laborscale 90 Uleselig Instrument groups used
24 Instrument bias compared to the reference interval Gender subgroup NORIP calculated reference limits Instrument mean (Max Min) in % of ref. interval B Haemoglobin (g/l) B Erc, volume fraction B Erythrocytes (10 12 /L) Low High Range Min Max Range Women ,7 143,4 0,7 2 Men ,7 143,4 0,7 2 Women 0,348 0,459 0,111 0,419 0,426 0,007 6 Men 0,395 0,5 0,105 0,419 0,426 0,007 7 Women 3,94 5,16 1,22 4,64 4,78 0,14 11 Men 4,25 5,71 1,46 4,64 4,78 0,14 10
25 Instrument bias compared to the reference interval Gender subgroup NORIP calculated reference limits Instrument mean (Max Min) in % of ref. interval Low High Range Min Max Range Erc MCV (fl) ,7 91,2 2,5 16 Erc MCH (pg) 27,1 33,3 6, ,8 0,8 13 Erc MCHC (g/l)
26 Instrument bias compared to the reference interval Gender subgroup NORIP calculated reference limits Instrument mean (Max Min) in % of ref. interval B Leukocytes (10 9 /L) B Thrombocytes (10 9 /L) Low High Range Min Max Range 3,47 8,81 5,34 5,4 5,9 0,5 9 Women Men All
27 NORIP reference intervals corrected for liquid anticoagulant Finnish reference intervals corrected for liquid anticoagulant (2 %) B Hb (g/l) Women Men Women and men B Erc, volume fraction B Erythrocytes (10 12 /L) 0,35-0,46 0,40-0,50 0,35-0,46 0,39-0,50 3,9-5,2 4,2-5,7 3,90-5,20 4,25-5,70 Erc MCV (fl) Erc MCH (pg) Erc MCHC (g/l) B Leukocytes (10 9 /L) B Thrombocytes (10 9 /L) ,5 8,8 3,4-8,
28 Preliminary haematology reference intervals Analyte Gender subgroup NORIP calculated NORIP suggested B Haemoglobin (mmol/l)* B Haemoglobin (g/l) B Erc, volume fraction B Erythrocytes (10 12 /L) Women 7,09 9,27 7,1 9,3 Men 8,12 10,30 8,1 10,3 Women Men Women 0,348 0,459 0,35 0,46 Men 0,395 0,500 0,40 0,50 Women 3,94 5,16 3,9 5,2 Men 4,25 5,71 4,2 5,7
29 Preliminary haematology reference intervals Analyte Gender subgroup NORIP calculated NORIP suggested Erc MCV (fl) 82,0 98, Erc MCH (fmol)* 1,64 2,02 1,6 2,0 Erc MCH (pg) 27,1 33, Erc MCHC (mmol/l)* 19,2 21,6 19,2 21,6 Erc MCHC (g/l)
30 Preliminary haematology reference intervals Analyte Gender subgroup NORIP calculated NORIP suggested B Leukocytes (10 9 /L) 3,47 8,81 3,5 8,8 Women B Thrombocytes (10 9 /L) # Men * Unit recommended by IFCC/IUPAC. # All For B Thrombocytes the partitioning rules suggest separate low reference limits for women and men but the working group tends to propose a common reference interval for both genders.
31
32 Thanks! These presentations are made with good support from Gunnar Nordin, Pål Rustad, Ari Lahti, Helge Erik Solberg and the working group for NORIP Haematology
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