Rev. Bras. Hematol. Hemoter. vol.38 número3

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revbrashematolhemoter.2016;38(3):225–239

w w w . r b h h . o r g

Revista

Brasileira

de

Hematologia

e

Hemoterapia

Brazilian

Journal

of

Hematology

and

Hemotherapy

Original

article

Assessment

of

blood

sample

stability

for

complete

blood

count

using

the

Sysmex

XN-9000

and

Mindray

BC-6800

analyzers

Sabrina

Buoro

a,∗

,

Tommaso

Mecca

a

,

Michela

Seghezzi

a

,

Barbara

Manenti

a

,

Lorenzo

Cerutti

a

_,

_Paola

_Dominoni

a

_,

_Gavino

_Napolitano

a

_,

_Stefano

_Resmini

a

_,

Alberto

Crippa

a

_,

_Cosimo

_Ottomano

b

_,

_Giuseppe

_Lippi

c

a_Hospital_Papa_Giovanni_XXIII,_Bergamo,_Italy

b_CAM_Centro_Analisi_Monza,_Monza,_Italy

c_Universitaà_degli_Studi_di_Verona,_Verona,_Italy

a

r

t

i

c

l

e

i

n

f

o

Articlehistory:

Received4April2016 Accepted30May2016 Availableonline23June2016

Keywords:

XN-9000 BC-6800 Samplestability Completebloodcount Pre-analyticalphase

a

b

s

t

r

a

c

t

Background:Differenthematologicalanalyzershavedifferentanalyticalperformancesthat

areoftenreflectedinthecriteriaforsamplestabilityofthecompletebloodcount.Thisstudy aimedtoassessthestabilityofseveralhematologicalparametersusingtheXN-9000Sysmex andBC-6800Mindrayanalyzers.

Methods:Theimpactofstorageatroomtemperatureand4◦_C_was_evaluated_after_2,_4,_6,

8,24,36and48husingtennormaland40abnormalbloodsamples.Thevariationfrom thebaselinemeasurementwasevaluatedbytheSteel–Dwass–Critchlow–Flignertestandby Bland–Altmanplots,usingqualityspecificationsandcriticaldifferenceasthetotalallowable variation.

Results:Red bloodcells andreticulocyteparameters (i.e. hematocrit,mean corpuscular

volume,mean corpuscularhemoglobinconcentration,redbloodcelldistributionwidth, immaturereticulocytefractions,low-fluorescencereticulocytes,middle-fluorescence retic-ulocytes,highfluorescencemononuclearcells)showedlessstabilitycomparedtoleukocyte andplateletparameters(exceptformonocytecountandmeanplateletvolume).Thebias forhematocrit,meancorpuscularvolume,meancorpuscularhemoglobinconcentration andredbloodcelldistributionwidthcoefficientofvariationwashigherthanthecritical differenceafter8husingbothanalyzers.

Conclusion: Bloodsamplesmeasuredwithbothanalyzersdonotshowanalytically

signifi-cantchangesinupto2hofstorageatroomtemperatureand4◦_C._However,_the_maximum

timeforanalysiscanbeextendedforupto8hwhenthebiasiscomparedtothecritical difference.

©2016Associac¸ ˜aoBrasileiradeHematologia,HemoterapiaeTerapiaCelular.Published byElsevierEditoraLtda.ThisisanopenaccessarticleundertheCCBY-NC-NDlicense (http://creativecommons.org/licenses/by-nc-nd/4.0/).

∗ _{Corresponding}_author_at_:_Clinical_Chemistry_Laboratory,_Papa_Giovanni_XXIII_Hospital,_Square_OMS,₁_-24127_Bergamo,_Italy.

E-mailaddress:[email protected](S.Buoro).

http://dx.doi.org/10.1016/j.bjhh.2016.05.010

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226 Introduction

Modern hematological analyzers not only enable accurate quantitativeandqualitativeassessmentofbloodcells,butalso provideavastarrayofhematologicalparametersthatmaybe usefulforthediagnosticandprognosticassessmentofmany bloodcelldisorders.Thevastmajorityoflaboratoryerrors(up to70%)emergefromthepre-analyticalphase.1_This_phase_is

influencedbyanumberofvariables,includingthepreparation ofthepatientbeforetesting,theproceduresusedtocollect andtransportthebiologicalspecimens,aswell asthetime andstorageconditionsofbloodsamplesbeforeanalysis.In particular,itwasrecentlyproventhatthestabilityofmany hematologicalparametersisstronglyinfluencedbythe stor-agetemperatureofthesampleandthetimeelapsedbetween collectionandanalysis.2–4

Anotherfactorthatmayhaveaninfluenceonthe stabil-ityofhematologicalparametersisthetechnologyusedbythe differenthematologicalanalyzers.3_Basically,_the_instruments

currentlyavailableonthemarketusedifferentmethodsand technologies toassess basicparameters such asred blood cell (RBC), platelet (PLT), total leukocyte (WBC) and leuko-cytesubclass[neutrophils(NE),lymphocytes(LY),monocytes (MO),eosinophils(EO),basophils(BA)]counts.5_It_is_for_this

reason that a more profound knowledge of the potential impactoftime and storagetemperatureofsamples before analysisshould be regarded asa mainstaytoincrease the qualityofhematologicaltestingandtoimprovetheclinical interpretationofdataobtainedwithdifferentanalyzersand techniques.2–4_Notably,_the_latest_generation_of_{hematological}

analyzersprovidesanumberofinnovativequantitativeand qualitativeparameters,suchastheenumerationofhigh fluo-rescencemononuclearcells(HFC)5–7_and_nucleated_red_blood

cell(NRBC)count,7_and_the_RBC_distribution_width_expressed

asastandarddeviation(RDW-SD)orcoefficientofvariation (RDW-CV).8_Moreover,_they_may_provide_platelet_distribution

width(PDW),plateletcrit(PCT),meanplateletvolume(MPV), percentage of large platelet (P-LCR) parameters,5,9,10 _along

withthereticulocytecount(RET)andimmaturereticulocyte fractions[IRF, high-fluorescence (HFR), middle-fluorescence (MFR)andlow-fluorescencereticulocytes(LFR)],allofwhich areusefulforthediagnosisandclassificationofanemiaorfor monitoringbonemarrowerythropoiesis.11,12

Theimportanceofverifyingthestabilityofthe aforemen-tionedparametersisnowunquestionableandpublisheddata aboutbloodsamplestabilitybeforeanalysisisscarceforthe newgeneration ofhematological analyzers.Therefore, this studyaimedtoassessandcomparethe stabilityofa num-ber of hematological parameters in normal and abnormal bloodsamplesmeasuredusingtwonovelanalyzers,XN-9000 (SysmexCo.,Kobe,Japan)andBC-6800(Mindray,Shenzhen, China),accordingtotheGuidelinesoftheInternational Coun-cilforStandardizationofHaematology(ICSH)4_and_the_Clinical

andLaboratoryStandardsInstitute(CLSI)DocumentH26-P2.13

Bloodsamples

The study population consisted of ten adult and osten-sibly healthy volunteers recruited from the laboratory

staff (five women, mean age 37.5±0.8 years and five males, mean age 35.0±7.4 years). All subjects were Cau-casian,hadnodiabetesmellitus,hypertensionandhadnot taken any medication for onemonthbefore the study.Six venous blood samplesfrom each subjectwere collected in K3-ethylenediaminetetraaceticacid(K3-EDTA)tubes(Becton

Dickinson, Franklin Lakes, NJ). All samples were analyzed immediatelyaftervenipuncture(i.e.,within30min).The anal-ysisoftheimpactofdifferentstoragetemperatureswasthen carriedoutbystoringthreebloodtubesfromeachindividualat roomtemperature(RT)andthreebloodtubesweredividedin sixaliquotsandstored(refrigerated)at4◦_C._Repeated

meas-ureswerethenperformedoneachsampleafter2h,4h,6h, 8h,24h,36hand48hofstorage.Anadditionalstudywas per-formedusing40routinesampleswithabnormalvalues,that is, containingatleastoneabnormalityofhemoglobin(Hb), platelet(PLT)orwhitebloodcell(WBC)countsor morpholog-icalalterations(i.e.,atleastonemorphologicalflagforWBC). CountabnormalitiesincludedHblowerthan70g/dL,PLTlower than100×109_/L_or_higher_than₄₀₀_×₁₀9_/L;_WBC_lower_than

1.00×109_/L_or_higher_than_12.00_×₁₀9_/L._{Hematological}

test-ingwasperformedimmediatelyuponarrivalinthelaboratory (i.e.,within30min)andtheneachsamplewasdividedinto8 aliquots,fourwerestoredat4◦_C_and_four_were_stored_at_RT.

Thetestswererepeatedafter4h,8h,24h36hand48hof stor-age.Allmeasures(i.e.,thebaselineandtherepeatedanalyses) wereperformedinduplicateandthefinalvaluewasexpressed asthemeanofthetwoanalysesateachtimepoint.

Studydesign

Thefollowingparameterswereassessedtocheckblood sam-ple stability: extended complete blood count (CBC) profile parameters, including all basic CBC parameters [RBC, Hb, hematocrit(HT)],meancorpuscularvolume(MCV),mean cor-puscularhemoglobin(MCH), mean corpuscularhemoglobin concentration (MCHC),RDW-SD, RDW-CVand NRBC. More-over,theextendeddifferentialcounts(DIFF)(includingWBC, NE,LY,MO,EO,BAandHFC),PLTprofileparameters(including PLT,PCT,MPV,PDWandP-LCR)andtheRETprofile(including RET,IRF,LFR,MFRandHFR)wereevaluated.

Themeasurementsatthedifferenttimepointswere con-comitantly performed withboth the XN-9000 and BC-6800 analyzers. The mean analytical characteristics of the two analyzers are summarized in Table 1. Briefly, the XN-9000 andtheBC-6000analyzersperforma5-partDIFF,RETcount, NRBCcount,withflagsappearinginthepresenceofabnormal results.7,14,15_Both_analyzers_use_a_combination_of_flow

cytom-etryandfluorescencewithlysingbuffersforleukocyteDIFF andidentification ofabnormalcells.Aseparatechannelfor NRBCassessmentisalsoavailableintheBC-6800.

The between-run imprecision ofboth the XN-9000 and BC-6800wasevaluatedaccordingtotheCLSIdocument EP5-A3,16 _by_analysis_in_duplicate_of_three_levels_(i.e., _level_1,₂

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227

Table1–XN-9000andBC-6800parameterswithoptimalperformanceBias%andCriticalDifference%(CD).

Parameters XN-9000 BC-6800 Optimal

performance Bias%

Critical Difference% onXN-9000

Critical Difference%on

BC-6800

Redbloodcell RBC RBC 0.9 9.1 9.3

Hemoglobin HGB HGB 0.9 7.9 8.1

Hematocrit HCT HCT 0.9 8.4 9.5

Meanvolume,redbloodcells MCV MCV 0.6 4.2 5.3

Meancorpuscularhemoglobin MCH MCH 0.7 5.0 5.4

Meancorpuscularhemoglobinconcentration MCHC MCHC 0.4 5.8 6.7

RBCdistributionwidth RDW-CV RDW-CV 0.9 9.8 9.9

RDW-SD RDW-SD NA NA NA

Nucleatedredbloodcell NRBC NRBC NA NA NA

Whitebloodcells WBC WBC 2.8 30.5 30.6

Neutrophil NEUT Neu 4.6 45.4 45.2

Lymphocyte LYMPH Lym 3.7 30.2 30.4

Monocyte MONO Mon 6.6 53.7 57.4

Eosinophil EO Eos 9.9 62.0 58.9

Basophil BASO Bas 7.7 77.9 88.1

Highfluorescencecells HFLC HFC NA NA NA

Platelet PLT PLT 3.0 26.1 26.2

Meanvolumeplatelet MPV MPV 1.2 12.3 12.6

PLTdistributionwidth PDW PDW NA NA NA

Plateletcrit PCT PCT NA NA NA

PLTlargercellratio P-LCR P-LCR NA NA NA

Reticulocyte RET RET 3.9 31.7 31.1

Immaturereticulocytefraction IRF IRF NA NA NA

Low-fluorescencereticulocyte LFR LFR NA NA NA

Medium-fluorescencereticulocyte MFR MFR NA NA NA

High-fluorescencereticulocyte HFR HFR NA NA NA

NA:notavailable.

relevantlocallegislationandwithpriorapprovaloftheLocal

EthicsCommittee.

Statisticalanalysis

Thesignificanceofthedifferenceoftheparametersobtained

in paired samples measured with the two analyzers was

estimated according to the Steel–Dwass–Critchlow–Fligner

test,withassessmentbytheHodges–Lehmannlocationshift

formultiple comparisons of means and medians between

differentgroups,afterverificationofvaluedistributionbythe

Shapiro–Wilktest.Statisticalsignificancewassetforp-values

<0.05. The results were then reported as delta variations

from baselineanalysis immediately aftercollection, asX

(TX−T0), where “X” isthe different timing and “0” is the

baselineresult.Percentagevariationsfromthebaselineresult

(T0)insampleswithstatisticallysignificantdifferenceswere

then analyzedusing Bland–Altmanplots (Bias%)and

com-paredwiththecurrentqualityspecificationsforoptimalbias

(OP-Bias%),17_that_is_calculated_using_{intra-individual}

biolog-icalvariability(CVi)andinter-individualbiologicalvariability (CVg)followingtheequation:OP-Bias%=0.250(CVi+CVg)1/2_.

Bias%wasalsocomparedwiththereferencechangevalues orcriticaldifference(CD)18,19_when_these_indices_were

avail-able. The CD percentage is the highest relative difference betweentwo consecutive measurements,that, ata chosen levelofprobability, maystillbeduetothecombinedeffect oftheanalytical(Va)andbiological(Vi)variations.Itisgiven bythe followingequation: CD%=K×(Va2₊_Vi2₎1/2_, _where _K

dependsonthechosenprobability.Thecomparisonbetween Bias% and CD% was performed only forthose parameters exhibitingastatisticallysignificantdifferencebetweenBias% and OP-Bias% throughoutthe study period. The statistical analysis was performed using Analyse-it software version 3.90.1(Analyse-itSoftwareLtd.;Leeds,UK).

Results

Overall, 2480measurements were performed withthe XN-9000andBC-6800analyzers.Allresultsobtainedinthenormal samples group were included in the statistical analysis, whereas160measurementswereperformedintheabnormal samplesgroup. Unfortunately,severalmeasurements could notbeperformedduetothelowresidualsamplevolumein this second groupofsamples. Theresultsofthese studies andtherelativevariationsaccordingtothedifferentstorage conditionsareshowninTables2–5.

Redbloodcellparameters

Themedianvaluesobtainedatbaseline(i.e.,T0)inthenormal sample group did not significantly differ between the two analyzersforalltheparameterstested,exceptforMCHand MCHC(Table2).Inthisgroupofnormalsamples,thevalues ofWBC,RBC,HB,MCHandNRBCwerefoundtobestableup to48hatRTand4◦_C_using_both_analyzers._Conversely,_the_HT

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Table2–SamplesstabilityofgroupofnormalsamplesforCBCandRETprofilesparameters.MedianHodges–Lehmannlocationshift(X_);_Bias%_(B%)_between_baseline (T0)andthetimepoint(2hupto48h)at4◦_C_and_room_temperature_(RT)_comparison_of_OP-Bias%_to_Critical_Difference_%_(CD).

Temp T0

medianvalue (95%CI)

X(TX−T0)Hodges–Lehmannlocationshift Xwith

p-value<0.0001 atthetime[h]

B%(95%CI)a timeofstability;

p-value B%vs.OP-Bias%;

Stableuntil[h]

B%(95%CI);

p-value B%vs.CD%; Stableuntil[h] 2h 4h 6h 8h 24h 36h 48h

WBC (109_/L)

XN-9000 RT 7.67(5.91to9.40) 0.03 0.05 −0.03 0.06 0.07 −0.08 −0.10 NS 1.4(−1.0to3.7);

p=0.2192;48h

NE

4◦_C _0.13 _0.02 _0.22 _0.14 _0.08 _0.10 ₋_0.03 _NS _3.0₍₋_1.4_to_7.5);

p=0.9109;48h

NE

BC-6800 RT 7.15(5.60to7.95) 0.02 0.00 0.04 0.14 0.16 0.19 0.01 NS −0.2(−2.5to2.0);

p=0.0105;48h

NE

4◦_C _0.01 _0.16 _0.02 _0.03 _0.03 _0.04 ₋_0.03 _NS _0.2₍₋_1.0_to_1.5);

p=0.0013;48h

NE

RBC (1012_/L)

XN-9000 RT 4.60(4.23to5.22) −0.01 −0.01 −0.02 0.01 0.02 0.01 0.00 NS −0.1(−0.6to0.5)

p<0.0001;48hb

NE

4◦_C _0.12 _0.10 _0.13 _0.15 _0.11 _0.16 _0.15 _NS _2.7_(0.9_to_4.5)

p=0.0522;48h

NE

BC-6800 RT 4.57(4.20to4.89) 0.00 0.09 0.05 −0.08 −0.06 −0.04 −0.04 NS −0.6(−3.9to2.7);

p=0.3477;48h

NE

4◦_C _0.06 _0.08 _0.06 _0.09 _0.09 _0.08 _0.09 _NS _1.6_(0.4_to_2.9);

p=0.2163;24h

NE

Hb(g/L)

XN-9000 RT 142(126to151) 0.10 0.20 0.10 0.20 0.30 0.20 0.10 NS 1.0(0.6to1.4)

p=0.5780;48h

NE

4◦_C _0.40 _0.35 _0.40 _0.30 _0.35 _0.35 _0.40 _NS _3.7_(2.0_to_5.4);

p<0.0001c

−0.6(−1.4to0.6); p<0.0001;48hd

BC-6800 RT 146(134to151) 0.00 0.00 0.00 −0.20 −0.15 −0.20 −0.10 NS −1.4(−4.3to1.5);

p=0.1119;48h

NE

4◦_C _0.20 _0.20 _0.10 _0.20 _0.20 _0.20 _0.20 _NS _1.3_(0.9_to_1.6);

p=0.0640;48h

NE

HT (%)

XN-9000 RT 41.6(37.2to43.7) −0.37 −0.63 −0.56 −0.40 2.11 3.25 6.12 48h 0.9(−1.6to3.5);

p=0.9747;8h

4.9(2.2to7.6); p<0.0001;24hd

4◦_C _0.25 _0.25 _0.40 _0.80 _0.95 _1.35 _1.90 _NS _2.3_(0.8_to_3.9);

p=0.0740;24h

4.7(3.3to6.4); p<0.0001;48hd

BC-6800 RT 41.4(40.2to44.1) −0.05 0.35 0.20 −0.60 2.15 3.90 6.00 48h 1.9(−1.0to4.8);

p=0.4791;8h

4.9(1.9to8.0) p<0.0001;24hd

4◦_C _0.20 _0.40 _0.00 _0.50 _0.90 _1.30 _2.00 _NS _1.6_(0.7_to_2.6);

p=0.1300;24h

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Table2–(Continued₎

Temp T0

medianvalue (95%CI)

Stableuntil[h]

B%(95%CI);

MCV (fL)

XN-9000 RT 87.45(86.2to90.5) −0.42 −1.24 −1.02 −1.27 3.51 6.10 13.1 24h 0.5(0.2to0.7);

p=0.3676;2h

−1.5(−2.1to

−0.8);

p<0.0001;8hd

4◦_C ₋_1.25 ₋_1.30 ₋_1.40 ₋_1.00 _0.10 _0.50 _2.00 _NS _0.3₍₋_0.2_to_0.8);

p=0.2932;24h

2.0(1.3to2.7); p<0.0001;48hd

BC-6800 RT 89.8(88.2to92.7) −0.25 −0.45 −0.50 −0.35 5.70 8.80 13.4 24h −0.2(−0.8to0.5);

p=0.1688;8h

−0.2(−0.8to0.5); p<0.0001;8hd

4◦_C ₋_1.50 ₋_1.40 ₋_1.55 ₋_1.10 ₋_0.15 _0.70 _1.55 _NS _0.8_(0.3_to_1.4);

p=0.3793;36h

2.1(1.3to2.9); p<0.0001;48hd

MCHa (pg)

XN-9000 RT 30.0(29.0to30.6) 0.30 0.40 0.50 0.40 0.50 0.30 0.30 NS 1.1(0.6to1.6);

p=0.1069;48h

NE

4◦_C _0.40 _0.40 _0.40 _0.30 _0.20 _0.10 _0.20 _NS _0.6_(0.1_to_1.1);

p=0.7434;48h

NE

BC-6800 RT 31.4(30.7to32.3) −0.20 −0.30 −0.30 −0.30 −0.30 −0.50 −0.30 NS 0.9(0.2to1.6);

p=0.5672;48h

NE

4◦_C ₋_0.30 ₋_0.30 ₋_0.10 ₋_0.30 ₋_0.30 ₋_0.30 ₋_0.30 _NS _0.8_(0.1_to_1.8);

p=0.8085;48h

NE

MCHCa (g/dL)

XN-9000 RT 34.2(33.4to34.4) 0.60 0.80 1.00 0.90 −0.80 −1.90 −4.20 4h 1.7(1.1to2.6);

p<0.0001c

−2.4(−3.5to −1.3); p<0.0001;8hd

4◦_C _1.10 _0.90 _1.10 _0.80 _0.40 _0.10 ₋_0.50 ₂_h _3.1_(2.5_to_3.7);

p<0.0001c

1.4(0.6to2.2); p<0.0001;48hd

BC-6800 RT 35.6(33.4to36.2) −0.10 −0.30 −0.10 −0.20 −2.30 −3.50 −4.90 24h −0.7(−1.2to−0.2); p=0.2126;8h

−0.7(−1.2to −0.2); p<0.0001;8hd

4◦_C _0.20 _0.10 _0.30 _0.00 ₋_0.40 ₋_0.60 ₋_1.10 ₄₈_h _0.9_(0.3_to_1.4);

p=0.1027;24h

2.9(2.1to3.7); p<0.0001;48hd

RDW-CV (%)

XN-9000 RT 12.9(12.7to13.1) 0.00 0.00 0.10 0.10 0.70 1.20 1.50 24h 0.9(0.4to1.4);

p=0.9785;8h

8.7(7.6to9.8); p<0.0001;48hd

4◦_C ₋_0.10 ₋_0.10 ₋_0.10 ₋_0.10 ₋_0.30 ₋_0.30 ₋_0.30 ₂₄_h _0.9_(0.6_to_1.3);

p=0.8055;8h

2.3(1.4to3.1); p<0.0001;48hd

BC-6800 RT 12.9(12.4to13.2) 0.00 0.20 0.30 0.30 1.20 1.60 1.90 24h 0.5(0.1to0.8);

p<0.0001;2hb

2.7(2.0to3.3); p<0.0001;8hd

4◦_C _0.10 _0.20 _0.20 _0.20 ₋_0.10 _0.00 ₋_0.10 _NS _0.3₍₋_0.3_to_1.0);

p=0.0912;48h

NE

RDW-SD (fL)

XN-9000 RT 41.4(39.4to43.2) −0.10 −0.70 −0.25 −0.55 4.40 7.00 12.0 24h 10.2(8.6to11.8)

CD%datanot available

4◦_C ₋_1.10 ₋_1.20 ₋_1.10 ₋_1.10 ₋_1.20 ₋_1.10 ₋_0.50 _NS _0.9₍₋_0.4_to_2.2)

BC-6800 RT 40.5(39.5to41.5) 0.00 0.50 0.70 0.90 7.15 10.1 12.8 24h 14.9(12.5to17.4)

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Temp T0

medianvalue (95%CI)

Stableuntil[h]

B%(95%CI);

NRBC (109_/L)

XN-9000 RT 0.0(0.0to0.0) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 NS 20.0(−31.7to71.7)

4◦_C _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _NS _0.0₍₋_42.9_to_42.9)

BC-6800 RT 0.0(0.0to0.0) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 NS 0.0(0.0to0.0)

4◦_C _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _NS _0.0_(0.0_to_0.0)

RETa (109_/L)

XN-9000 RT 48.1(35.1to60.3) 0.63 −0.89 −1.05 −1.50 −3.71 −0.59 −1.77 NS 2.5(−1.5to6.6);

p=0.4964;48h

NE

4◦_C _1.01 _1.15 _0.36 _0.08 ₋_0.07 _0.27 ₋_0.11 _NS _0.3₍₋_3.9_to_4.4);

p=0.0840;48h

NE

BC-6800 RT 37.4(26.8to44.4) 0.85 1.55 0.30 −1.10 0.45 −1.20 −2.60 NS 6.1(0.3to12.0);

p=0.4344;36h

9.8(4.5to15.1); p<0.0001;48hd

4◦_C _0.00 _0.50 ₋_1.30 _0.25 _1.30 _1.65 _3.45 _NS _5.0_(1.2_to_8.8);

p=0.5394;36h

10.7(6.2to15.2); p<0.0001;48hd

IRFa (%)

XN-9000 RT 9.3(6.9to10.3) −0.81 −0.49 −1.08 −1.37 −1.94 −1.85 −2.07 24h −25.1(−38.7to−11.5)

4◦_C _0.15 ₋_0.98 ₋_0.42 ₋_0.96 ₋_0.64 ₋_0.66 _0.45 _NS _4.0₍₋_5.3_to_13.4)

BC-6800 RT 3.4(2.5to4.3) 0.20 0.20 0.20 −0.20 −0.85 −1.35 −2.00 36h −90.2(−110.8to−69.6)

4◦_C _0.0 ₋_0.80 ₋_0.60 ₋_0.70 _0.10 _0.50 _0.90 _NS _22.1_(4.4_to_39.6)

LFRa (%)

XN-9000 RT 90.7(89.7to93.1) 0.81 0.48 1.08 1.37 1.94 1.85 2.07 24h 2.1(1.0to3.2)

4◦_C ₋_0.16 _0.98 _0.42 _0.96 _0.64 _0.66 ₋_0.45 _NS ₋_0.5₍₋_1.4_to_0.4)

BC-6800 RT 96.5(95.7to97.5) −0.20 −0.20 −0.20 0.20 0.85 1.35 2.00 36h 2.1(1.5to2.7)

4◦_C _0.00 _0.80 _0.60 _0.70 ₋_0.10 ₋_0.50 ₋_0.90 _NS ₋_0.8₍₋_1.3_to₋_0.2)

MFRa (%)

XN-9000 RT 8.4(6.5to8.9) −0.72 −0.43 −1.02 −1.20 −1.58 −1.70 −1.68 24h −23.7(−37.6to−9.7)

4◦_C _0.15 ₋_0.78 ₋_0.48 ₋_0.82 ₋_0.77 ₋_0.66 _0.04 _NS ₋_0.9₍₋_11.2_to_9.5)

BC-6800 RT 3.4(2.5to4.3) 0.20 0.20 0.20 −0.20 −0.85 −1.35 −2.00 36h −90.2(−67.9to−35.5)

4◦_C _0.00 ₋_0.80 ₋_0.60 ₋_0.70 _0.10 _0.50 _0.90 _NS _22.0_(4.4_to_39.7)

HFRa (%)

XN-9000 RT 0.9(0.6to1.3) 0.00 0.00 −0.10 −0.10 −0.30 −0.10 −0.30 NS −51.1(−79.2to−23.0)

4◦_C _0.10 ₋_0.15 _0.10 ₋_0.10 _0.10 _0.10 _0.40 _NS _46.8_(21.2_to_72.4)

BC-6800 RT 0.0(0.0to0.0) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 NS 0.0(0.0to0.0)

4◦_C _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _NS _0.0_(0.0_to_0.0)

WBC:whitebloodcells;RBC:redbloodcell;RT:Roomtemperature;Hb:hemoglobin;HT:hematocrit;MCV:meancorpuscularvolume;MCH:meancorpuscularhemoglobin;MCHC:meancorpuscular hemoglobinconcentration;RDW-CV:distribution-coefficientofvariation;RDW-SD:RBCdistributionwidth-standarddeviation;NRBC:nucleatedredbloodcell;RET:reticulocyte;IRF:immature reticulocytefraction;LFR:low-fluorescencereticulocyte;MFR:medium-fluorescencereticulocyte;HFR:high-fluorescencereticulocyte;NS:Xnotsignificantthroughoutthestudyperiod;NE:not evaluated;Temp:Temperature.

a _Parameters_with_median_value_a_T0_significant_difference_between_two_analyzer_in_the_same_samples_with_p_<_0.0001.

b _Bias%_(between_baseline_T0_and_the_time_point_X)_is_lower_than_OP-Bias%.

c _Bias%_(between_baseline_T0_and_the_time_point_X)_is_always_higher_than_OP-Bias%.

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231

Table3–Samplesstabilityongroupofpathologicalsamplesfor:CBCandRETprofilesparameters.Median

Hodges–Lehmannlocationshift(X_);_Bias%_(B%)_between_baseline_(T0)_and_the_time_point₍₄_h_up_to₈_h)_at₄◦_C_and_room

temperature(RT)comparisonofOP-Bias%toCriticalDifference%(CD).

Temp T0medianvalue (95%CI)

X(TX−T0) Hodges–Lehmann

locationshift

Xwith

Stableuntil[h]

B%(95%CI);

p-value B%vs.CD%; Stableuntil[h]

4h 8h

WBC (109_/L)

XN-9000 RT 5.3(4.3to7.2) 0.00 −0.10 NS −0.8(−6.8to5.2);

p=0.2229;8h

NE

4◦_C _8.6_(6.5_to_10.1) _0.06 _0.04 _NS _0.6₍₋_4.5_to_5.6); p=0.37625;8h

NE

BC-6800 RT 5.4(4.1to7.1) 0.05 −0.10 NS 2.3(−0.9to5.5);

p=0.3274;8h

NE

4◦_C _8.5_(6.4_to_9.9) _0.20 ₋_1.22 _NS _1.8_(0.7_to_2.9); p=0.0774;4h

18.4(7.2to29.7);

p<0.0001;8ha

RBC (1012_/L)

XN-9000 RT 4.8(3.8to5.6) 0.01 −0.16 NS −4.0(−17.8to9.8);

p=0.4582;8h

NE

4◦_C _3.6_(3.5_to_3.9) ₋_0.02 ₋_0.02 _NS ₋_0.4₍₋_2.1_to_1.2); p=0.1161;8h

NE

BC-6800 RT 4.6(3.7to5.3) −0.02 −0.01 NS 0.3(−1.1to1.8);

p=0.4194;8h

NE

4◦_C _3.5_(3.3_to_3.8) ₋_0.01 _0.26 _NS _8.5₍₋_0.5_to_17.5);

p=0.0945;8h

NE

Hb (g/L)

XN-9000 RT 141.0(123.0to152.0) 0.10 −0.70 NS −5.2(−19.7to9.2); p=0.3370;8h

NE

4◦_C _113.0_(101.0_to_116.0) _0.00 _0.00 _NS _0.5₍₋_0.4_to_1.3); p=3067;8h

NE

BC-6800 RT 141.0(123.0to154.0) −0.10 −0.20 NS −0.8(−4.3to2.7); p=0.3108;8h

NE

4◦_C _111.0_(103.0_to_117.0) _0.00 _1.30 _NS _1.6_(0.07_to_3.12); p=0.3546;8h

NE

HT (%)

XN-9000 RT 40.7(35.3to46.5) 0.40 −1.40 NS −3.5(−17.4to10.3);

p=0.5044;8h

NE

4◦_C _33.9_(31.5_to_35.5) _0.00 _0.00 _NS _0.1₍₋_0.7_to_0.9);

p=0.0614;8h

NE

BC-6800 RT 39.7(34.5to44.3) 0.00 0.00 NS 1.3(−0.3to2.9);

p=0.5982;8h

NE

4◦_C _32.6_(30.6_to_34.4) _0.00 _0.02 _NS _8.6₍₋_0.5_to_17.7);

p=0.0921;8h

NE

MCV (fL)

XN-9000 RT 84.8(84.0to88.4) 0.60 0.30 NS 0.5(0.1to0.8);

p=0.4860;8h

NE

4◦_C _89.9_(87.7_to_92.5) _0.40 _0.40 _NS _0.5₍₋_0.3_to_1.4);

p=0.8606;8h

NE

BC-6800 RT 85.8(84.5to88.7) 0.60 0.80 NS 0.5(0.4to0.7);

p=0.3379;4h

1.0(0.8to1.2); p<0.0001;8ha

4◦_C _92.4_(87.1_to_93.5) _0.20 _0.10 _NS _0.1_(0.0_to_0.3);

p<0.0001;8hb

NE

MCH (pg)

XN-9000 RT 28.9(27.3to29.8) 0.07 −0.33 NS 1.2(0.1to2.4);

p=0.3425;8h

NE

4◦_C _29.6_(27.2_to_30.9) _0.20 _0.00 _NS ₋_0.1₍₋_1.0_to_0.8); p=0.0961;8h

NE

BC-6800 RT 30.3(28.9to31.4) 0.00 0.10 NS −1.1(−4.0to1.8);

p=0.2119;8h

NE

4◦_C _31.3_(28.2_to_32.4) _0.10 _0.85 _NS _0.20₍₋_0.07_to_0.05); p=0.1530;4h

(8)

232

locationshift

Xwith

Stableuntil[h]

B%(95%CI);

4h 8h

MCHC (g/dL)

XN-9000 RT 33.6(32.5to34.5) −0.10 −0.60 NS 0.5(−0.3to1.2);

p=0.8634;4h

NE

4◦_C _32.9_(32.4_to_33.5) _0.02 ₋_0.20 _NS _0.6_(0.3_to_0.9); p=0.1932;8h

NE

BC-6800 RT 35.3(34.7to35.6) −0.35 −0.40 NS −2.1(−5.0to0.8);

p=0.812;8h

NE

4◦_C _34.1_(33.7_to_34.3) _0.10 _0.85 ₈_h _0.0₍₋_0.3_to_0.3); p<0.0001b_;₄_h

3.8(1.6to6.1); p<0.0001;8ha

RDW-CV (%)

XN-9000 RT 13.3(13.1to15.9) 0.20 0.35 NS 2.8(−9.7to15.4);

p<0.7469;8h

NE

4◦_C _14.2_(13.4_to_14.6) _0.10 _0.10 _NS _0.7_(0.1_to_1.2);

p=0.4190;8h

NE

BC-6800 RT 13.4(13.0to15.6) 0.30 0.50 NS 2.1(1.8to2.4);

p<0.0001c

3.6(2.9to4.2); p<0.0001;8ha

4◦_C _13.7_(13.4_to_14.1) _0.10 _0.25 _NS _0.9_(0.5_to_1.2);

p=0.9609;4h

2.1(1.2to2.9); p<0.0001;8ha

RDW-SD (fL)

XN-9000 RT 42.3(41.1to44.3) 1.00 1.85 NS _p3.6₌_0.6544;(−9.0to₈16.2);_h

4◦_C _44.2_(43.1_to_48.4) _0.50 _0.60 _NS _1.2₍₋_0.2_to_2.5);

p=0.6629;8h

BC-6800 RT 41.5(40.3to43.3) 1.20 2.00 8h 4.3(3.6to5.1)

4◦_C _42.7_(41.9_to_47.8) _0.40 _0.90 _NS _2.0_(1.0_to_2.9)

NRBC (109_/L)

XN-9000 RT 0.0(0.0to0.0) 0.00 0.01 8h 103.3(41.9to164.8)

4◦_C _0.0_(0.0_to_0.0) _0.01 _0.02 ₄_h _129.9_(95.5_to_164.4)

BC-6800 RT 0.0(0.0to0.0) 0.00 0.00 NS 78.6(26.4to130.8)

4◦_C _0.0_(0.0_to_0.0) _0.00 _0.02 ₈_h _124.0_(78.5_to_169.6)

RET (109_/L)

XN-9000 RT 53.1(46.3to72.1) 0.11 −3.39 NS −4.4(−24.8to16.1);

p=0.3996;8h

NE

4◦_C _54.0_(46.0_to_68.1) _1.20 _1.50 _NS _3.0₍₋_0.1_to_6.1);

p=0.5427;8h

NE

BC-6800 RT 41.5(31.9to55.2) 2.30 3.55 NS 7.3(3.3to11.4);

p=0.0912;8h

NE

4◦_C _52.2_(41.1_to_60.1) _1.14 _3.45 _NS _5.4₍₋_5.2_to_16.0);

p=0.7652;8h

NE

IRF (%)

XN-9000 RT 6.6(4.1to7.9) 0.00 0.60 NS 6.1(−28.3to40.5)

CD%datanot available 4◦_C _15.5_(11.5_to_20.5) _0.20 _0.10 _NS ₋_0.1₍₋_6.5_to_6.3)

BC-6800 RT 1.7(0.9to2.5) −0.20 −0.30 NS −8.5(−28.5to11.5)

4◦_C _8.4_(5.0_to_12.9) ₋_0.60 ₋_0.80 _NS ₋_21.5₍₋_44.7_to_1.7)

LFR (%)

XN-9000 RT 93.4(92.1to95.9) 0.00 −0.60 NS −0.7(−6.5to5.0)

4◦_C _84.6_(79.5_to_88.5) ₋_0.20 ₋_0.10 _NS ₋_0.7₍₋_2.6_to_1.1)

BC-6800 RT 98.3(97.5to99.1) 0.20 0.30 NS 0.1(−0.5to0.6)

4◦_C _91.7_(87.1_to_95.0) _0.60 _0.80 _NS _0.7_(0.2_to_1.2)

MFR (%)

XN-9000 RT 6.1(4.0to6.9) 0.15 0.60 NS 6.8(−22.5to36.2)

4◦_C _12.0_(10.6_to_13.8) ₋_0.55 ₋_0.40 _NS ₋_5.6₍₋_10.7_to₋_0.5)

BC-6800 RT 1.7(0.9to2.5) −0.20 −0.30 NS −8.9(−28.9to11.1)

4◦_C _8.0_(5.0_to_11.6) ₋_0.30 ₋_0.60 _NS ₋_18.9₍₋_42.5_to_4.8)

HFR (%)

XN-9000 RT 0.5(0.1to1.0) 0.00 0.00 NS 6.9(−56.2to70.0)

4◦_C _2.8_(1.4_to_7.9) _0.40 _0.30 _NS _19.0₍₋_2.1_to_40.1)

BC-6800 RT₄_◦_C _0.00.0(0.0_(0.0to_to0.0)_1.2) 0.00_0.00 0.00_0.00 _NSNS 2.6(−1.7to6.9) −38.8(−70.2to−7.4)

WBC:whitebloodcells;RBC:redbloodcell;RT:Roomtemperature;Hb:hemoglobin;HT:hematocrit;MCV:meancorpuscularvolume;MCH: meancorpuscularhemoglobin;MCHC:meancorpuscularhemoglobinconcentration;RDW-CV:distribution-coefficientofvariation;RDW-SD: RBCdistributionwidth-standarddeviation;NRBC:nucleatedredbloodcell;RET:reticulocyte;IRF:immaturereticulocytefraction;LFR: low-fluorescencereticulocyte;MFR:medium-fluorescencereticulocyte;HFR:high-fluorescencereticulocyte;NS:Xnotsignificantthroughoutthe studyperiod;NE:notevaluated;Temp:Temperature.

a _Bias%_(between_baseline_T0_and_the_time_point_X)_is_lower_than_CD%.

b _Bias%_(between_baseline_T0_and_the_time_point_X)_is_lower_than_OP-Bias%.

(9)

rev

bras

hema

tol

hemoter.

2

0

1

6;

3

8(3)

:225–239

233

Table4–Samplesstabilityofgroupofnormalsamplesfor:DIFFandPLTprofilesparameters.MedianHodges–Lehmannlocationshift(X);Bias%(B%)betweenbaseline (T0)andthetimepoint(2hupto48h)at4◦_C_and_room_temperature_(RT)_comparison_of_OP-Bias%_to_Critical_Difference%_(CD).

X(TX−T0)Hodges–Lehmannlocationshift Xwithp<0.0001 atthetime[h]

Stableuntil[h]

B%(95%CI);

2h 4h 6h 8h 24h 36h 48h

NE (109_/L)

XN-9000 RT 4.4(3.5to4.7) 0.01 0.08 −0.02 0.00 0.21 0.06 0.00 NS 0.5_p₌_0.0273;(−3.1to₄₈4.1);_h NE

4◦_C _0.07 _0.07 _0.09 _0.06 _0.04 _0.14 _0.13 _NS _2.5₍₋_3.4_to_8.3);

p=0.4509;48h

NE

BC-6800 RT 4.0(3.0to4.6) 0.17 0.12 0.09 0.21 0.40 0.27 0.42 NS 3.5(0.9to6.1);

p=0.3950;48h

NE

4◦_C _0.02 _0.10 _0.00 _0.05 _0.23 _0.34 _0.51 _NS _11.0_(7.1_to_14.8);

p=0.1137;24h

NE

LY (109_/L)

XN-9000 RT 2.3(1.7to2.5) −0.02 −0.02 −0.07 −0.05 0.00 0.03 0.05 NS 1.1(−1.7to3.9);

p=0.0626;48h

NE

4◦_C ₋_0.03 _0.01 ₋_0.02 _0.00 ₋_0.09 ₋_0.06 ₋_0.11 _NS _7.6_(3.1_to_12.1); p=0.0885;48h

NE

BC-6800 RT 2.0(1.6to2.3) 0.01 0.00 0.02 0.02 0.02 0.00 −0.01 NS 1.0(−2.7to4.7);

p=0.1480;48h

NE

4◦_C ₋_0.01 ₋_0.02 ₋_0.03 _0.00 ₋_0.13 _0.12 _0.24 _NS _0.3₍₋_2.9_to₋_3.47); p=0.0372;8h

16.4(11.1to21.7);

p<0.0001;48hb

MOa (109_/L)

XN-9000 RT 0.6(0.6to0.9) 0.00 0.00 −0.01 −0.01 −0.12 −0.15 −0.03 NS 16.0(5.0to27.0);

p=0.0884;48h

NE

4◦_C _0.02 _0.00 _0.02 _0.02 ₋_0.01 ₋_0.06 ₋_0.17 _NS _8.8_(1.94_to₋_15.7);

p=0.5053;24h

28.1(19.1to37.1); p<0.0001;48hb

BC-6800 RT 0.5(0.4to0.7) 0.00 0.00 0.01 0.00 −0.04 −0.04 −0.12 48h 12.3(5.1to19.4);

p=0.1128;36h

33.2(24.7to41.6); p<0.0001;48hb

4◦_C _0.02 _0.03 _0.02 _0.01 ₋_0.01 ₋_0.07 ₋_0.13 ₄₈_h _3.4₍₋_1.5_to_8.2);

p=0.1816;24h

36.7(28.6to44.8); p<0.0001;48hb

EO (109_/L)

XN-9000 RT 0.2(0.1to0.3) 0.00 0.01 0.02 0.03 −0.02 −0.05 −0.01 NS 14.5(2.0to−26.9);

p=0.4507;48h

NE

4◦_C _0.00 _0.00 _0.01 _0.00 _0.01 _0.00 _0.00 _NS _0.4₍₋_11.7_to_12.5);

p=0.1175;48h

NE

BC-6800 RT 0.1(0.1to0.3) 0.01 0.00 0.01 0.01 0.00 0.01 0.00 NS _p6.1₌_0.2908;(−1.2to₄₈13.4);_h NE

4◦_C _0.01 _0.01 _0.00 _0.01 _0.01 _0.01 _0.01 _NS _10.01_(0.9_to_19.3);

p=0.966;48h

NE

BA (109_/L)

XN-9000 RT 0.1(0.0to0.1) 0.00 0.00 0.00 0.00 0.01 0.01 0.01 NS 20.2(−4.5to44.9);

p=0.3032;48h

NE

4◦_C _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _0.01 _NS _26.1_(1.3_to_50.9);

p=0.1362;48h

(10)

234

rev

bras

hema

tol

hemoter.

2

0

1

6;

3

8(3)

:225–239

Table4–(Continued)

X(TX−T0)Hodges–Lehmannlocationshift Xwithp<0.0001 atthetime[h]

Stableuntil[h]

B%(95%CI);

BC-6800 RT 0.0(0.0to0.0) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 NS _p3.8₌_0.5552;(−9.9to₄₈17.4);_h NE

4◦_C _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _NS _10.5₍₋_10.8_to_30.9); p=0.7777;48h

NE

HFC (109_/L)

XN-9000 RT 0.0(0.0to0.0) 0.00 0.00 0.00 0.00 −0.01 −0.01 −0.01 NS −88.3(−134.8to−41.9)

CD%datanot available 4◦_C _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _NS ₋_26.7₍₋_73.9_to_20.6)

BC-6800 RT 0.0(0.0to0.0) 0.00 0.00 0.00 0.00 0.00 0.00 0.00 NS −28.7(−84.4to27.1) 4◦_C _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _0.00 _NS ₋_58.0₍₋_112.1_to₋_3.9)

PLT (109_/L)

XN-9000 RT 262.0_299.0)(231.0to −1.0 3.0 −3.0 −1.0 −19.0 −15.0 −10.0 NS 5.2_p₌_0.1847;(1.9to8.5);₄₈_h NE

4◦_C ₋_14.5 ₋_16.5 ₋_20.5 ₋_19.0 ₋_21.0 ₋_19.5 ₋_23.0 _NS _6.0_(4.0_to_8.0); p<0.0001c

9.5(6.1to12.8); p<0.0001;48hb

BC-6800 RT 250.0_279.0)(227.0to 9.0 7.0 12.0 6.5 3.0 7.0 5.0 NS 4.0_p₌_0.0045;(1.2to6.8);₄₈_h NE

4◦_C _4.0 _3.0 _4.0 ₋_1.0 ₋_3.0 ₋_2.0 ₋_5.0 _NS ₋_0.6₍₋_2.9_to_1.7);

p=0.4497;48h

NE

MPVa_(fL) XN-9000

RT

10.8(10.2to11.3) 0.40 0.50 0.60 0.60 1.30 1.50 0.90 24h 3.7(2.9to4.5);p<0.0001c 9.2(6.9to1.0); p<0.0001;48hb

4◦_C _0.00 ₋_0.10 _0.20 _0.10 _0.50 _0.60 _0.90 ₄₈_h _0.8₍₋_0.4_to_1.5);

p=0.1719.8h

5.7(4.3to7.0); p<0.0001;48hb

BC-6800 RT 9.5(9.1to9.9) 0.60 0.80 1.00 0.80 0.80 0.80 1.10 4h 6.1(4.9to7.2);p<0.0001c 8.8_p_<_0.05;(5.7to₂₄11.9);_hb

4◦_C _0.60 _0.50 _0.70 _0.60 _0.90 _1.00 _1.20 ₂₄_h _1.3_(1.1_to_1.4);

p=0.5307;48h

1.3(1.1to1.4); p<0.0001;48hb

PDWa (fL)

XN-9000 RT 12.5(11.8to14.0) 0.70 0.90 1.00 0.90 1.30 3.40 2.05 24h 19.7(14.7to24.3)

4◦_C _0.20 ₋_0.20 _0.40 _0.00 _1.00 _1.30 _2.00 ₄₈_h _13.7_(10.9_to_16.4)

BC-6800 RT₄◦_C _11.30)10.95(10.40to 0.80_1.00 1.10_0.60 _1.251.20 1.10_1.30 _1.601.30 1.20_1.60 _2.101.50 ₂₄24_hh _1.71.3_(1.1(0.5_toto2.0)_2.3)

PCTa_(%) XN-9000

RT

0.28(0.25to0.33) 0.01 0.01 0.01 0.01 0.02 0.03 0.02 NS 7.2(3.9to10.5)

4◦_C ₋_0.02 ₋_0.02 _0.02 ₋_0.02 ₋_0.01 ₋_0.01 _0.00 _NS ₋_2.3₍₋_4.8_to_0.3)

BC-6800 RT₄◦_C 0.2(0.2to0.3) 0.02_0.02 0.02_0.01 0.03_0.02 _0.010.03 0.02_0.02 _0.030.02 _0.300.03 _NSNS _11.615.4_(8.5(10.5_toto_14.7)20.4)

P-LCRa (%)

XN-9000 RT 32.3(27.4to36.0) 3.65 4.20 5.00 4.45 10.70 12.00 7.40 24h 30.3(23.2to37.3)

4◦_C _0.05 ₋_0.70 _1.25 _0.40 _3.55 _5.00 _6.95 ₄₈_h _20.1_(18.1_to_22.2)

BC-6800 RT 23.4(19.8to25.5) 3.80 5.00 6.00 5.30 5.05 5.00 6.80 6h 30.1(19.1to41.0)

4◦_C _3.95 _3.00 _4.65 _4.20 _6.40 _6.90 _8.25 ₂₄_h _33.0_(28.0_to_38.0)

NE:neutrophil;LY:lymphocyte;MO:monocyte;RT:Roomtemperature;EO:eosinophil;BA:basophil;HFC:HIGHfluorescencecells;PLT:platelet;MPV:meanvolumeplatelet;PDW:PLTdistributionwidth; PCT:plateletcrit;P-LCR:PLTlargercellratio;NS:Xnotsignificantthroughoutthestudyperiod;NE:notevaluated;Temp:Temperature.

a _Parameters_with_median_value_a_T0_significant_difference_between_two_analyzer_in_the_same_samples_with_p_<_0.0001. b _Bias%_(between_baseline_T0_and_the_time_point_X)_is_lower_than_CD%.

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Table5–SamplesstabilityofgroupofpathologicalsamplesforDIFFandPLTprofilesparameters.Median

Hodges–Lehmannlocationshift(X_);_Bias%_(B%)_between_baseline_(T0)_and_the_time_point₍₄_h_up_to₈_h)_at₄◦_C_and_room

temperature(RT)comparisonofOP-Bias%toCriticalDifference%(CD).

locationshift

Xwith

Stableuntil[h]

B%(95%CI);

4h 8h

NE (109_/L)

XN-9000 RT 2.9(2.3to4.5) −0.03 −0.09 NS 0.2(−8.9to9.3);

p=0.3146;8h

NE

4◦_C _5.8_(4.2_to_7.6) _0.07 _0.03 _NS _1.0₍₋_1.2_to_3.1); p=0.015;8h

NE

BC-6800 RT 3.0(2.3to4.6) 0.01 −0.03 NS 1.2(−1.8to4.3);

p=0.0324;8h

NE

4◦_C _5.9_(4.3_to_7.7) _0.12 ₋_0.87 _NS _2.2_(0.8_to_3.6); p=0.0017;4h

18.7(6.9to30.6);

p<0.0001;8ha

LY (109_/L)

XN-9000 RT 1.6(0.9to2.2) 0.02 −0.02 NS −2.8(−33.2to27.5);

p=0.6514;8h

NE

4◦_C _1.2_(1.0_to_1.8) ₋_0.01 _0.00 _NS ₋_1.7₍₋_15.1_to_11.8);

p=0.4237;8h

NE

BC-6800 RT 1.6(0.9to2.2) −0.02 −0.04 NS 4.4(0.2to8.5);

p=0.7316;8h

NE

4◦_C _1.2_(0.9_to_1.7) ₋_0.01 ₋_0.17 _NS _0.9₍₋_1.9_to₋_3.7);

p=0.0559;4h

13.3(4.7to21.4); p<0.0001;8ha

MO (109_/L)

XN-9000 RT 0.5(0.4to0.7) 0.01 −0.02 NS −4.9(−17.0to7.2);

p=0.0618;8h

NE

4◦_C _0.8_(0.6_to_0.9) _0.01 ₋_0.02 _NS _2.3₍₋_4.2_to_8.7);

p=0.1814;8h

NE

BC-6800 RT 0.4(0.3to0.5) 0.01 −0.01 NS 5.5(−0.6to11.7);

p=0.7159;8h

NE

4◦_C _0.6_(0.5_to_0.7) _0.02 ₋_0.13 _NS _3.0₍₋_0.2_to_6.2);

p=0.0279;4h

25.1(7.9to42.3); p<0.0001;8ha

EO (109_/L)

XN-9000 RT 0.1(0.0to0.4) 0.01 0.01 NS 43.4(−16.0to102.7);

p=0.2463;8h

NE

4◦_C _0.1_(0.1_to_0.1) _0.01 _0.02 _NS _8.9_(1.7_to_16.0);

p=0.7689;4h

31.5(16.2to46.7); p<0.0001;8ha

BC-6800 RT 0.1(0.0to0.4) 0.01 0.01 NS 29.0(−6.0to65.2);

p=0.0716;4h

29.6(−6.2to 65.3); p<0.05;8ha

4◦_C _0.1_(0.1_to_0.2) _0.01 _0.00 _NS _7.3₍₋_19.7_to_34.4);

p=0.8439;8h

NE

BA (109_/L)

XN-9000 RT 0.0(0.0to0.0) 0.00 0.01 NS 32.7(−8.2to73.6);

p=0.2113;8h

NE

4◦_C _0.0_(0.0_to_0.0) _0.00 _0.01 _NS _15.5₍₋_2.0_to_33.0);

p=0.3737;4h

30.3(17.8to42.8); p<0.0001;8ha

BC-6800 RT 0.0(0.0to0.0) 0.00 0.00 NS 0.7(−15.9to17.3);

p=0.3829;8h

NE

4◦_C _0.0_(0.0_to_0.0) _0.00 _0.00 _NS _7.1₍₋_16.1_to_30.4);

p=0.9604;8h

NE

HFC (109_/L)

XN-9000 RT 0.0(0.0to0.0) 0.00 0.00 NS −40.0(−125.8to45.8)

4◦_C _0.0_(0.0_to_0.0) _0.00 _0.00 _NS ₋_21.3₍₋_54.8_to_12.3)

BC-6800 RT 0.0(0.0to0.0) 0.00 0.00 NS 33.3(−26.1to92.8)

4◦_C _0.0_(0.0_to_0.0) _0.00 _0.00 _NS _4.3₍₋_59.0_to_67.5)

PLT (109_/L)

XN-9000 RT 264(211to309) 0.8 16.4 NS 4.0(−7.6to15.5);

p=0.8577;8h

NE

4◦_C ₂₃₇₍₁₈₈_to₂₇₀₎ _1.1 ₋_1.7 _NS ₋_0.6₍₋_5.7_to_4.4);

p=0.1548;8h

NE

BC-6800 RT 249(209to286) 0.5 −8.0 NS −3.7(−7.5to0.1);

p=0.7113;8h

NE

4◦_C ₂₃₅₍₁₇₈_to₂₇₆₎ ₋_2.0 ₋_9.0 _NS ₋_8.2₍₋_17.7_to_1.3);

p=0.2672;8h

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236

locationshift

Xwith

Stableuntil[h]

B%(95%CI);

4h 8h

MPV (fL)

XN-9000 RT 10.7(10.2to11.3) 0.10 0.05 NS 0.3(−4.0to4.6);

p=0.6606;8h

NE

4◦_C _11.2_(10.6_to_12.0) _0.11 _0.30 _NS _1.0_(0.3_to_1.6); p=0.4789;4h

2.6%(1.4to3.8);

p<0.0001;8ha

BC-6800 RT 9.7(8.7to10.2) 0.30 0.40 NS 2.6(1.5to3.7);

p<0.0001b

3.7%(2.7to4.7); p<0.0001;8ha

4◦_C _9.9_(9.4_to_10.5) _0.20 _0.50 _NS _2.7_(2.1_to_3.3);

p<0.0001b

4.7%(3.7to5.9); p<0.0001;8ha

PDW (fL)

XN-9000 RT 13.0(11.3to15.1) 0.29 0.10 NS 0.3(−7.4to8.1)

4◦_C _13.2_(12.2_to_15.1) _0.20 _0.60 _NS _4.6_(2.1_to_7.2)

BC-6800 RT 15.9(15.4to16.1) −0.05 0.00 NS 0.0(−0.6to0.5)

4◦_C _15.8_(15.7_to_16.1) _0.00 _0.00 _NS _0.1₍₋_0.5_to_0.7)

PCT (%)

XN-9000 RT 0.3(0.3to0.3) 0.00 0.01 NS 4.4(−6.1to14.9)

4◦_C _0.3_(0.2_to_0.3) _0.00 _0.01 _NS _2.1₍₋_3.4_to_7.5)

BC-6800 RT 0.2(0.2to0.2) 0.00 0.00 NS −0.1(−4.0to3.8)

4◦_C _0.2_(0.2_to_0.3) _0.00 _0.00 _NS ₋_4.2₍₋_13.8_to_5.4)

P-LCR (%)

XN-9000 RT 31.3(25.9to36.5) 1.36 0.72 NS 1.7(−9.7to13.1)

4◦_C _34.3_(29.4_to_40.3) _0.74 _2.24 _NS _6.5_(3.7_to_9.4)

BC-6800 RT 23.8(15.5to29.7) −0.20 −0.30 NS 9.2(6.7to11.7)

4◦_C _24.5_(21.0_to_29.4) _1.40 _3.95 _NS _12.8_(8.8_to_16.7)

NE:neutrophil;RT:Roomtemperature;LY:lymphocyte;MO:monocyte;EO:eosinophil;BA:basophil;HFC:highfluorescencecells;PLT:platelet; MPV:meanvolumeplatelet;PDW:PLTdistributionwidth;PCT:plateletcrit;P-LCR:PLTlargercellratio;NS:Xnotsignificantthroughoutthe studyperiod;NE:notevaluated;Temp:Temperature.

a _Bias%_(between_baseline_T0_and_the_time_point_X)_is_lower_than_CD%.

b _Bias%_(between_baseline_T0_and_the_time_point_X)_is_always_higher_than_OP-Bias%.

collectioninsamplesstoredatRT(butnotinthosestoredat 4◦_C)_when_measured_with_both_analyzers._The_comparison

oftheBias%atdifferenttimepoints(TX)withtheOP-Bias% showedthatHTisstableatroomtemperatureforupto8h usingboththeXN-9000(Bias%:0.9)andBC-6800(Bias%:1.9), whereastheBias%forHTinsamplesstoredatRTremained lower than the CD% for up to 24h. The same analysis in samplesstoredat4◦_C _showed_good _stability_for_up_to₂₄_h

aftercollectionwhencomparedwiththeOP-Bias%,whereas theBias%alwaysremainedlowerthantheCD%throughout thestudyperiod(i.e.,upto48h)usingbothanalyzers.

ThevaluesofMCVandRDW-SDdisplayedsignificant differ-encesafter24hofstorageatRT,whereastheresultsremained substantiallyunchangedforupto48haftercollectionin sam-plesstoreda4◦_C._{Interestingly,}_the_Bias%_of_MCV_exceeded

theOP-Bias%after2hofstorageatRTwiththeXN-9000and after8hofstorageatRTwiththeBC-6800,respectively.The Bias%ofMCVwaslowerthantherelativeCD%forupto8h ofstorageusingbothanalyzersatRT.Atvariance,theBias% alwaysremainedlowerthanCD%throughoutthestudyperiod insamplesstoredat4◦_C._The_RDW-CV_exhibited_significant

variationsafter24hfromcollectionusingbothanalyzersat RT,whereassignificantdifferenceswereobservedafter24h ofstorageinsamplesstoredat4◦_C_using _the_XN-9000 _but

notwiththeBC-6800(Table2).Whencomparedwiththe OP-Bias%,RDW-CVvalueswerefoundtobestableforupto8h atbothRTand4◦_C_using_the_XN-9000,_and_for_up_to₂_h_at

RTwiththeBC-6800.TheBias%ofRDW-CVwasalwayslower than the OP-Bias%for up to48h ofstorageusing the BC-6800.

Atvariancewithpreviousparameters,thevaluesofMCHC displayed a specific and instrument-dependent variation. Morespecifically,significantdifferenceswereobservedafter 4hofstorageatRTand2hofstorageat4◦_C_with_the_XN-9000.

Accordingly,theBias%exceededtheOP-Bias%at2hofstorage atbothtemperatures,whereastheBias%didnotexceedthe CD%forupto8hofstorageatRTandforupto48hofstorage at4◦_C._As_regards_MCHC_values_obtained_with_the_BC-6800,

significantdifferenceswerefoundafter24hofstorageatRT andatthe48htimepointafterstorageat4◦_C._The_OP-Bias%

wasexceededafter8hofstorageatRTand24hofstorage at4◦_C,_whereas_the_Bias%_remained_lower_than_the_CD%_for

upto8hofstorageatRTandforupto48hofstorageat4◦_C

(Table2).

Alltheseparametersappearedtobesubstantiallystablefor upto8hatbothRTand4◦_C_in_the_abnormal_samples_group

usingbothanalyzers.Themostrelevantexceptionsare sum-marizedinTable3.Specifically,theNRBCmeasuredwiththe XN-9000showedsignificantvariations8haftercollectionin samplesstoredatRTand4haftercollectioninthosestored at4◦_C,_whereas_significant_variations_of_NRBC_measured_with

theBC-6800couldonlybeobservedafter8hofstorageat4◦_C.