Prezentace aplikace PowerPoint

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A<= /div>

c<= /div>

ŠUM

λ1

λ2

λ1

λ2

c<= span style=3D'font-family:"Times New Roman";mso-ascii-font-family:"Times New Ro= man"; mso-hansi-font-family:"Times New Roman";font-size:76%;color:#FF33CC; mso-color-index:5;position:relative;top:.37em;mso-text-raise:-25%'>1<c<= span style=3D'font-family:"Times New Roman";mso-ascii-font-family:"Times New Ro= man"; mso-bidi-font-family:"Times New Roman";mso-hansi-font-family:"Times New Ro= man"; font-size:76%;color:#FF33CC;mso-color-index:5;position:relative;top:.37em; mso-text-raise:-25%'>2

Metoda λ1 má vyšší analytickou

citlivost= než λ2 (tgα1 > tgα2)=

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Mez detekce (limit of detectio= n- LD)

nnejmenš= ;í množství analytu, které může být ve vzorku kvalitativně dokázáno

n = ; LD =3D blank + 3*sbl = ; &n= bsp; SD =3D s

n =

nhodnota vyjadřuje schopnost analytického systému rozlišit přístrojový šum od analytického signál= u

n<= span style=3D'text-shadow:auto'>číselně definuje přístrojový šum

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D

ŠUM

c

<= span style=3D'mso-spacerun:yes'> X ± sbl = ;

<= span style=3D'mso-special-format:lastCR;display:none'>

L= d =3D průměr blanku + 3*sbl

λ 1

λ 2

c0 c0 c0

Ld λ<= /span>1

Ld λ<= /span>2

=

=

<= span style=3D'mso-special-format:lastCR;display:none'>

<= span style=3D'mso-special-format:lastCR;display:none'>

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=

výpočet   → experiment → validace s RM*

nNej= nižší množství analytu číselně stanovitelné s přijatelnou = hodnotou nejistoty odlišitelné hodnotou signálu od p&#= 345;ístrojového šumu (signálu blanku).


        = ;            &n= bsp;       Lq =3D blank + 6*sbl       (někteří autoři 10* sbl)

        = ;            &n= bsp;





§V praxi realizována hodnotou nejnižšího kalibrátoru= = a nižší koncentrac= e jsou hodnoceny: „< Lq“

        = ;            &n= bsp;      * RM =3D referenční materiál=

Mez stanovitelnosti (Lq)

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Analytick&e= acute; metody o vyšší analytické citlivosti vykazují nižší meze detekce a stanovitelnosti

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Analytická specifičnost = metody

<= span style=3D'font-family:Arial;text-shadow:auto'>nVlastnost metody, která vyjadřuje, jak stanovení analytu ovlivňuje (ruší, interferuje) přítomnost dalších látek endogenního či exogenního původu ve vyšetřovaném biologickém materiálu

= nAnalytická interference látek

<= nobr>= nAnalytická interference samotné matrice

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Analytická specifično= st

nAnalytická interference způsobuje systematickou chybu měření

n

nHb, lipidy, bilirubin, antikoagulancia při = bch analýze sér

nkřížové ICH reak= ce (aldosteron vs kortison)

nkyselina askorbová při ch= emické analýze moči

nléky podobné struktury při to= xikologické analýze

n

= nAnalytickou specifičnost zjistíme porovnáním s referenční metodou (hodnotíme % odchylku s rostoucím přídavkem interferentu vůči povoleným mezím)

n
nInterference klinické= (diuretika, indukce EE P450,…)

n

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Analytická interference bilirubinu
(od koncentrace 0,5 g/l)

D

Přidaný bilirub= in g/l

0 0,1 0,2 0,3 0,4 0,5 0,6

x

+ 2SD

- 2SD

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Výpočet výtěžnosti (recovery)

n         Naměřená hodnota cholesterolu

n

nv séru (hodnota cílová):        = ;   5,86 mmol/l

n<= span style=3D'text-shadow:auto'>ve frakci LDL + HDL LP:       4,25 mmol/l

n<= span style=3D'text-shadow:auto'>ve frakci CHM + VLDL LP:   1,18 mmol/l

        = ;            &n= bsp;    100 * (frakce 1 + frakce 2= )         543

Výtěžnost<= span style=3D'font-family:"Times New Roman";mso-ascii-font-family:"Times New Ro= man"; mso-hansi-font-family:"Times New Roman";font-size:114%;text-shadow:auto; color:white;mso-color-index:1'> =3D -------------------------- =3D ------- =3D 93%

        = ;            &n= bsp;         sérová hodnota        = ;   5,86

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Linearita metody

<= span style=3D'font-size:75%;text-shadow:auto'>nRozsah kalibrační funkce s lineárním průběhem,

<= span style=3D'font-size:75%;text-shadow:auto'>nJde o určitou <= b>část<= b> (úsek) každé funkce!

<= span style=3D'font-size:75%;text-shadow:auto'>nVýsledek se kvantifikuje směrnicí přímky y =3D B*x + A= 3;

<= span style=3D'font-size:75%;color:yellow;mso-color-index:3;text-shadow:auto; visibility:hidden'>n

<= span style=3D'font-size:75%;color:yellow;mso-color-index:3;text-shadow:auto'>nOvěře= ní linearity

sadou RM v příslušné koncentračn&iacut= e; řadě

= ; - vytvo= 45;ením ředící řady z nativního materiálu vhodným

= ; rozpouštědlem (negativní vzorek, fyziologický

= ; roztok, 0. kalibrátor,…)

= ; &n= bsp;

= ; Správnost ředění != !

= ; Shoda průběhů kalibrátor vs biologický materi&aacut= e;l!!=

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Pracovní rozsah=

U lineární= ch metod, resp. při měření v lineární oblasti dána dolní a horn&i= acute; mez linearity (záruka spolehlivosti stanovení analytu) <= /span>

U <= /span>nelineárních metod o pracovním rozsahu metody rozhoduje rozsah hodnot komerčních kalibrátorů = ;

<= nobr>n

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= ; Validita (platnost, spolehlivost)

= ; Celková analytická chyba = (TE, total error)

Rozdíl výsledku měření od hodnoty veličiny v RM.

<= /div>

Kvantifikace systematické= chyby (<= /span>bi= as) a náhodné = ;

chyby analýzy (CVa) měření (statist. spolehlivost 95%):

= ; &n= bsp; Odhad analytické nejistoty (Uc)&= #13;

= ;

= ; Uc =3D 2*CVa + = |b| <= /div>

= ;

= ; Uc =3D 2*Ö(CVa2 + |b| 2)

<= span style=3D'mso-special-format:lastCR;display:none'>

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Content-Location: file:///C:/A37AC9E1/kontrola_soubory/slide0233.htm Content-Transfer-Encoding: quoted-printable Content-Type: text/html; charset="us-ascii" Prezentace aplikace PowerPoint

Absolutní (definitivní) metoda

gravimetrie, odměrné titrace, coulometrie = ;

= ; Bias 0% CVa 0%

= ; &n= bsp; = Referenční metoda

důkladně ustanovený, prostudovaný, definovaný

certifikovaný a validovaný měříc&iac= ute; postup

HPLC-MS, sekvenování DNA, HPCE, AAS

= ; Bias 0% CVa ~ 1%

= ; &n= bsp; Standardizovan= á

= ; rutinní metoda

Bias dle návaznosti CVa 1-10%

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Kalibrát= ory v klinické biochemii

nPrimárn&iac= ute; kalibrátor – analyt v rozpouštědle

= ; &n= bsp; = ; - výhoda: čistota, stabilita

= ; &n= bsp; = ; - nevýhoda: nezohledňují matrici

= ; použi= tí pro: kalibrace referenčních metod

n<= /span>

<= span style=3D'font-size:88%;color:yellow;mso-color-index:3;text-shadow:auto'>nSekundárn&i= acute; kalibrátor – analyt v matricovém základu

(komutabilní s biologickými materiály) s hodno= tami

určenými referenčními metodami či odvozené z

certifikovaných RM; <= /span>pro rutinní kalibrace (a EHK)

n<= /span>

<= span style=3D'font-size:88%;color:yellow;mso-color-index:3;text-shadow:auto'>nMultikalibrá= ;tor <= /span>– sekundární kalibrátor obsahující více

= ; &n= bsp; rozpuštěných analytů

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Komutabilita

=3D shoda vlastností referenčn&iacu= te;ho materiálu (kalibrátoru, kontroly)= s reálnými vzorky v průbě= hu měření

… aby materiál byl co nejvíc nativní

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Požadavky na referenční materiál

Komutabilita

Stabilita

Neinfekčnost

Návaznost laboratorních m= etod =

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Kdy kalibrovat a rekalibrovat?=

nNový stroj, nová aplikace, nové analytické podmínky

nPožadavek výrobce (četnost dle stability &= #269;inidel, nové šarže)

nPo opravě a údržbě stroje

nVelká systematická chyba měření (EHK) =

Základem kalibrace je = preciznost: uskladnění, pipety, rozpouštědla, doba rozpouš= ;tění, dublety, čtení pokynů a informací výrobce kalibrátoru

K =3D 1,25

<= span style=3D'mso-special-format:lastCR;display:none'>

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Kontrola kvality
(pr&a= acute;ce v laboratoři)

nVnitřní kontrola kvality (VKK, IQC)

Informuje o stabilitě analytického systému

Základem dlouhodobé hodnocení CVa (%)

nExterní kontrola kvality (EHK, EQA)

systematické chybě

Základem hodnocen&ia= cute; bias (a výpočet z skore)

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Vnitřní kontrola kvali= ty

/VKK, IQC/

nZajišťuje stabilitu celé analytické fáze laboratorního měření a spolehlivost výsledku&= #13;

n2 materiály nejlépe 2x denně na začátku směny

nReferenční meze kontrolních materiálů

nPoužívat stejnou předem otestovanou šarži po 6-12M

Testován&iacut= e; v sérii 20 měření (průměr, bias, CV= a, ±3s)

n1x měsíčně průměr a SEM

nPrůběžně diagr= am Levey-Jenningsův

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Levey-Jenningsův graf VKK= 3;

+3s

+2s

+1s

-1s

-2s

-3s<= /div>

Série (měsíční průměry měření)

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Westgardova kontrolní pravidla VKK<= span style=3D'mso-spacerun:yes'> 13s, 22s, R3s, 10x

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Série (měsíční průměry měření)

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Content-Transfer-Encoding: quoted-printable Content-Type: text/html; charset="us-ascii" Prezentace aplikace PowerPoint

Externí hodnocení kvality <= b>/EHK, EQA/

nMezilaborator= ní porovnání analytických metod, reagencií a přístrojů

nanalýzy identických vzorků

nHodnocení analytů <= span style=3D'text-shadow:auto;color:white;mso-color-index:1'>v delší časové ose

(nejčastěji= 1-4x ročně)

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<= span style=3D'mso-special-format:lastCR;display:none'>

Klinick&ea= cute; laboratoře ČR

Organiz= átor EHK

* Hodnocení účastníka

* Identifikace chyb

* Hodnocení časových trendů

* Doklad o účasti

* Certifikát úspěšnosti

Matematické zpracování výsledků= = ;

<= span style=3D'mso-special-format:lastCR;display:none'>

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Hodnocení úspěšnosti v EHK

DL Cíl= ová hodnota = HL

Umax

UTV

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Jak se hodnotí úspěšnost laboratoře?

<= /div>

<= span style=3D'mso-spacerun:yes'> - dle odchylky od cílové hodnoty

<= span style=3D'font-family:Arial;font-size:75%;color:yellow;mso-color-index:3; text-shadow:auto'>n<= span style=3D'font-family:Arial;mso-ascii-font-family:Arial;mso-hansi-font-fami= ly: Arial;font-size:75%;text-shadow:auto;color:yellow;mso-color-index:3'>C&= iacute;lová hodnota RM zí= ;skaná změřením analytu

= ; referen= 9;ní metodou (např. glukosa)

n<= span style=3D'font-family:Arial;mso-ascii-font-family:Arial;mso-hansi-font-fami= ly: Arial;font-size:75%;text-shadow:auto;color:yellow;mso-color-index:3'>C&= iacute;lová hodnota RM zí= ;skaná v několika expertních

laboratořích metodou doporučenou IFCC (enzym= y)

= ;

<= span style=3D'font-family:Arial;font-size:75%;color:#CCFFCC;text-shadow:auto'><= span style=3D'mso-special-format:bullet;color:yellow;mso-color-index:3;position= :absolute; left:-3.99%;top:.3em;font-family:Wingdings;font-size:75%'>n<= span style=3D'font-family:Arial;mso-ascii-font-family:Arial;mso-hansi-font-fami= ly: Arial;font-size:75%;text-shadow:auto;color:#CCFFCC'>Cílová hodnota= získaná jako průměr účastníků EHK

= ; &n= bsp; používajících tutéž analytickou metodu <= /div>

= ; (hormony met= odou RIA firmy Amersham)

(zde nelze hodnotit správnost metody,= 3;

jen porovnatelnost !!!)

------=_NextPart_01CA626C.98BFB110 Content-Location: file:///C:/A37AC9E1/kontrola_soubory/slide0357.htm Content-Transfer-Encoding: quoted-printable Content-Type: text/html; charset="us-ascii" Prezentace aplikace PowerPoint

= ; Validita (platnost, spolehlivost)

= ; Celková analytická chyba = (TE, total error)

Rozdíl výsledku měření od hodnoty veličiny v RM.

<= /div>

Kvantifikace systematické= chyby (<= /span>bi= as) a náhodné = ;

chyby analýzy (CVa) měření (statist. spolehlivost 95%):

= ; &n= bsp; Odhad analytické nejistoty (Uc)&= #13;

= ;

= ; Uc =3D 2*CVa + = |b| <= /div>

= ;

= ; Uc =3D 2*Ö(CVa2 + |b| 2)

<= span style=3D'mso-special-format:lastCR;display:none'>

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Content-Location: file:///C:/A37AC9E1/kontrola_soubory/slide0353.htm Content-Transfer-Encoding: quoted-printable Content-Type: text/html; charset="us-ascii" Prezentace aplikace PowerPoint

Příklad hodnocení Uc= (%)

= nOdhaduje nejistoty rutinního měření glukosy v séru dle výsledků VKK a <= span style=3D'font-family:"Times New Roman";mso-ascii-font-family:"Times New Ro= man"; mso-hansi-font-family:"Times New Roman";font-size:114%;text-shadow:auto; color:yellow;mso-color-index:3'>EHK

1. Reprodukovatelnost při VKK: CVa =3D 1,2%

2. Pravdivost (správnost, bias) při EHK: b =3D 3,0%

= ; CVb =3D |b| / Ö3 =3D 1,7% (3… n-dní měření)

3. Nejistota hodnoty glukosy v RM: UTV =3D 0,5%&= #13;

4. Uc =3D 2x Ö= ; (CVa2 + CVb2 + UTV2) =3D 4,3%

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Hodnocení Uc dle z-skóre v rámci EHK

Předpoklady výpočtu:

- n&a= acute;va= znost na referen&#= 269;n<= /span>&i= acute; metodu: b&re= g;0<= /span>

- je známa dlouhodobá SD metody v dané laboratoři

- laboratoř je účastníkem EHK (a dána cílová hodn= ota TV)

= ; &n= bsp; = ; &n= bsp; = ; &n= bsp; = ; &n= bsp;

= ; &n= bsp; = ; naměřená hodnota - TV

= ; z skóre =3D -------------------------------

= ; &n= bsp; = ; &n= bsp; SD metody

Z-skóre >3 =3D hrubá chyba

Z-skóre £ 1 =3D metoda naprosto v pořádku

nejobjektiv= nější ukazatel dlouhodobé kvality výsledků labor. metody =

<= /div>

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Co patří do znaků proveditelnosti ?

nČas odezvy mezi příjmem materiálu a <= /span>výdejem výsledku po celých= 24 hodin

nPř&iac= ute;mé a nepřímé náklady a platba ZP =

nAnalytické, technické, ekonomické a log= istické zabezpečení laboratoře&n= bsp;

= ; viz laboratorní dokumentace

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Analytické cíle

nRozvoj technologie – přesnějš&iacut= e; analýzy

n (↓ CV) <= /div>

nNové referenční metody a komutabilnější materiály – vyšší náv= aznost (↓ bias)

nNové statistické metody a výpočetní technika

nSnižování nejistoty v pre- a post-ana= lyt. Fázi

nSnižován&i= acute; nejistoty vs. cena vyšetření

n = ; &n= bsp; = … existují smysluplné hranice ?

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Analytické cíle – odvození složek nejistot z biologických rozptylů analytů

CVi =3D intraindividuální biologický rozptyl

        = ; vypočítaný tzv. chronobiologickými studiemi



CVg =3D interindividuální (skupinový) rozptyl =

<= /span>

CVc =3D celkový biologický rozp= tyl =3D CVi + CVg

<= /span>

        = ; CVa &= pound; 0,5 * CVi        = ;   |b| £ 0,25 * CV<= span style=3D'font-family:"Times New Roman";mso-ascii-font-family:"Times New Ro= man"; mso-hansi-font-family:"Times New Roman";font-size:67%;text-shadow:auto; color:yellow;mso-color-index:3;position:relative;top:.37em;mso-text-raise: -25%'>c

<= /span>

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Zákl= adem standardní laboratorní práce je především:

1.

1.Pečlivě číst příbalové informace výrobce

2.

2.Provádět kontrolu kvality (KK, QC)
3.

3.Vést záznamy o technickém stavu přístrojů

4.

4.Vést pečlivou laboratorní dokumentaci

5.

5.

5.

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document.oncontextmenu=3Dparent._CM; document.onkeydown =3D _KDH; document.ondragstart=3DCancel document.onselectstart=3DCancel self.focus() } function Redirect( frmId ) { var str=3Ddocument.location.hash,idx=3Dstr.indexOf('#'), sId=3DGetSldId() if(idx>=3D0) str=3Dstr.substr(1); if( window.name !=3D frmId && ( sId !=3D str) ) { obj =3D GetObj("Main-File") window.location.href=3Dobj.href+"#"+sId return 1 } return 0 } var MHTMLPrefix =3D CalculateMHTMLPrefix(); function CalculateMHTMLPrefix() { if ( document.location.protocol =3D=3D 'mhtml:') { href=3Dnew String(document.location.href) Start=3Dhref.indexOf('!')+1 End=3Dhref.lastIndexOf('/')+1 if (End < Start) return href.substring(0, Start) else return href.substring(0, End) } return ''; } function GetTags(base,tag) { if(g_supportsPPTHTML) return base.all.tags(tag); else return base.getElementsByTagName(tag); } function UpdNtsPane(){ if(frames["PPTNts"]) PPTNts.location.replace( MHTMLP= refix+GetHrefObj( gCurSld ).mNtsHref ) } function UpdNavPane( sldIndex ){ if(gNavLoaded) PPTNav.UpdNav() } function UpdOtNavPane(){ if(gOtlNavLoaded) PPTOtlNav.UpdOtlNav() } function UpdOtlPane(){ if(gOtlLoaded) PPTOtl.UpdOtl() } function SetHasNts( fVal ) { if( gHasNts !=3D fVal ) { gHasNts=3DfVal UpdNavPane() } } function ToggleOtlText() { gOtlTxtExp=3D!gOtlTxtExp UpdOtlPane() } function ClearMedia() { if (PPTSld.pptSound) PPTSld.pptSound.loop =3D 0; } function FullScreen() { if ( PPTSld.g_animUseRuntime ) PPTSld.document.body.pause(); ClearMedia(); var href =3D ( document.location.protocol =3D=3D 'mhtml:') ? FULLSCR_HREF = : FULLSCR_HREF+"#"+GetHrefObj(gCurSld).mSldHref; if(PPTNav.event.ctrlKey) { var w =3D (window.screen.availWidth * 1.0) / 2.0 var h =3D w * (PPTSld.g_origH * 1.0) / PPTSld.g_origW win =3D window.open( MHTMLPrefix+href,null,"toolbar=3D0,resizable=3D1,top= =3D0,left=3D0," + "width=3D"+ w + ",height=3D" + h ); if( win.document.body && PPTSld.g_animUseRuntime ) win.document.body.PPTSldFrameset=3Dwindow; } else { win =3D window.open( MHTMLPrefix+href,null,"fullscreen=3Dyes" ); if( win.document.body && PPTSld.g_animUseRuntime ) win.document.body.PPTSldFrameset=3Dwindow; } } function ToggleVNarration() { rObj=3DPPTSld.document.all("NSPlay") if( rObj && !PPTSld.g_animUseRuntime ) { if( (rObj.playState =3D=3D 1)||(rObj.playState =3D=3D 0) ) rObj.Play() else if( rObj.playState =3D=3D 2 ) rObj.Pause() else return; } else if( PPTSld.g_animUseRuntime ) { narObj =3D PPTSld.document.all("narrationID") if( narObj ) narObj.togglePause() } } function GetCurSldNum() { obj=3DGetHrefObj(gCurSld) if( obj.mOrigVis =3D=3D 1 ) return obj.mSldIdx else return gCurSld } function GetNumSlds() { if( GetHrefObj(gCurSld).mOrigVis =3D=3D 1 ) return GetSldList().mNumVisSlds; else return GetSldList().mList.length } function GetSldNum( href ) { for(ii=3D0; ii 1 ) PopSldList(); else if( !IsFramesMode() ) { if( gLoopCont ) GoToFirst() else EndShow() } } function GoToPrevSld() { ii=3DgCurSld-1 if( ii > 0 ) { obj=3DGetHrefObj(ii) while ( obj && ( obj.mVis =3D=3D 0 ) && ( ii>0 ) ) obj=3DGetHrefObj(--ii) if( ii =3D=3D 0 ) ii=3D1 GoToSldNum(ii) } } function GoToFirst(){ GoToSld( GetHrefObj(1).mSldHref ) } function GoToLast() { ii=3DGetSldList().mList.length if( ii !=3D gCurSld ) GoToSld( GetHrefObj(ii).mSldHref ) } function GoToSldNum( num ) { if( PPTSld.event ) PPTSld.event.cancelBubble=3Dtrue obj =3D GetHrefObj( num ) obj.mVis=3D1 gPrevSld=3DgCurSld gCurSld =3D num; PPTSld.location.replace(MHTMLPrefix+obj.mSldHref) if( IsFramesMode() ) { UpdNavPane(); UpdOtlPane(); UpdNtsPane() } } function GoToSld( href ) { if( PPTSld.event ) PPTSld.event.cancelBubble=3Dtrue GetHrefObj( GetSldNum(href) ).mVis=3D1 PPTSld.location.replace(MHTMLPrefix+href) } function SldUpdated( id ) { if( id =3D=3D GetHrefObj(gCurSld).mSldHref ) return gPrevSld=3DgCurSld gCurSld=3DGetSldNum(id) if( IsFramesMode() ) { UpdNavPane(); UpdOtlPane(); UpdNtsPane() } } function PrevSldViewed(){ GoToSld( GetHrefObj(gPrevSld).mSldHref ) } function HasPrevSld() { return ( gIsEndShow || ( gCurSld !=3D 1 && GetHrefO= bj( gCurSld-1 ).mVis =3D=3D 1 )||( GetCurSldNum() > 1 ) ) } function HasNextSld() { return (GetCurSldNum() !=3D GetNumSlds()) } function CloseWindow() { if( HideMenu() ) return; var event =3D PPTSld.event; if( !IsFramesMode() && event && (event.keyCode=3D=3D27 || event.keyCode=3D= =3D32 || event.type=3D=3D"click" ) ) window.close( self ); CatchNumKeys( self, event ); } function Unload() { gIsEndShow=3D0; } function SetupEndShow() { gIsEndShow=3D1; PPTSld.document.body.scroll=3D"no"; PPTSld.document.onkeypress=3DCloseWindow; PPTSld.document.onclick=3DCloseWindow; PPTSld.document.oncontextmenu=3D_CM; } function EndShow() { if( IsFramesMode() ) return if( PPTSld.event ) PPTSld.event.cancelBubble=3Dtrue doc=3DPPTSld.document var dir =3D doc.body.dir if( dir !=3D "rtl" ) dir =3D "ltr"; doc.open() doc.writeln('

' + ENDSHOW_MESG + '
') doc.close() } function SetSldVisited(){ GetSldList().mList[gCurSld-1].mVisited=3Dtrue } function IsSldVisited(){ return GetSldList().mList[gCurSld-1].mVisited } function hrefList( sldHref, visible, advDelay, advClk ) { this.mSldHref=3D this.mNtsHref =3D sldHref this.mOrigVis=3D this.mVis =3D visible this.mVisited=3D false this.mAdvDelay=3D advDelay this.mAdvOnClk=3D advClk } function SldList(arr,curSld,fEnd) { this.mCurSld =3D curSld; this.mList =3D new Array(); var idx =3D 1; for(ii=3D0;ii 0) { PushSldList(sldList,fEnd); gCurSld =3D 1; } else if( PPTSld.event ) PPTSld.event.cancelBubble=3Dtrue } function PushSldList(arr,fEnd) { var ii =3D gSldStack.length; gSldStack[ii] =3D new SldList(arr,gCurSld,fEnd); GoToSld( gSldStack[ii].mList[0].mSldHref ); } function PopSldList() { if (gSldStack[gSldStack.length-1].fEndShow) EndShow() else { gCurSld =3D gSldStack[gSldStack.length-1].mCurSld; gSldStack[gSldStack.length-1] =3D null; gSldStack.length--; var sldList =3D gSldStack[gSldStack.length-1]; GoToSld( sldList.mList[gCurSld - 1].mSldHref ); } } var custShowList=3Dnew Array(); function ImgBtn( oId,bId,w,action ) { var t=3Dthis t.Perform =3D _IBP t.SetActive =3D _IBSetA t.SetInactive=3D _IBSetI t.SetPressed =3D _IBSetP t.SetDisabled=3D _IBSetD t.Enabled =3D _IBSetE t.ChangeIcon =3D null t.UserAction =3D action t.ChgState =3D _IBUI t.mObjId =3D oId t.mBorderId=3D bId t.mWidth =3D w t.mIsOn =3D t.mCurState =3D 0 } function _IBSetA() { if( this.mIsOn ) { obj=3Dthis.ChgState( gHiliteClr,gShadowClr,2 ) obj.style.posTop=3D0 } } function _IBSetI() { if( this.mIsOn ) { obj=3Dthis.ChgState( gFaceClr,gFaceClr,1 ) obj.style.posTop=3D0 } } function _IBSetP() { if( this.mIsOn ) { obj=3Dthis.ChgState( gShadowClr,gHiliteClr,2 ) obj.style.posLeft+=3D1; obj.style.posTop+=3D1 } } function _IBSetD() { obj=3Dthis.ChgState( gFaceClr,gFaceClr,0 ) obj.style.posTop=3D0 } function _IBSetE( state ) { var t=3Dthis GetObj( t.mBorderId ).style.visibility=3D"visible" if( state !=3D t.mIsOn ) { t.mIsOn=3Dstate if( state ) t.SetInactive() else t.SetDisabled() } } function _IBP() { var t=3Dthis if( t.mIsOn ) { if( t.UserAction !=3D null ) t.UserAction() if( t.ChangeIcon ) { obj=3DGetObj(t.mObjId) if( t.ChangeIcon() ) obj.style.posLeft=3Dobj.style.posLeft+(t.mCurState-4)*t.mWidth else obj.style.posLeft=3Dobj.style.posLeft+(t.mCurState-0)*t.mWidth } t.SetActive() } } function _IBUI( clr1,clr2,nextState ) { var t=3Dthis SetBorder( GetObj( t.mBorderId ),clr1,clr2 ) obj=3DGetObj( t.mObjId ) obj.style.posLeft=3Dobj.style.posLeft+(t.mCurState-nextState)*t.mWidth-obj= .style.posTop t.mCurState=3DnextState return obj } function TxtBtn( oId,oeId,action,chkState ) { var t=3Dthis t.Perform =3D _TBP t.SetActive =3D _TBSetA t.SetInactive=3D _TBSetI t.SetPressed =3D _TBSetP t.SetDisabled=3D _TBSetD t.SetEnabled =3D _TBSetE t.GetState =3D chkState t.UserAction =3D action t.ChgState =3D _TBUI t.mObjId =3D oId t.m_elementsId=3D oeId t.mIsOn =3D 1 } function _TBSetA() { var t=3Dthis if( t.mIsOn && !t.GetState() ) t.ChgState( gHiliteClr,gShadowClr,0,0 ) } function _TBSetI() { var t=3Dthis if( t.mIsOn && !t.GetState() ) t.ChgState( gFaceClr,gFaceClr,0,0 ) } function _TBSetP() { if( this.mIsOn ) this.ChgState( gShadowClr,gHiliteClr,1,1 ) } function _TBSetD() { this.ChgState( gFaceClr,gFaceClr,0,0 ) this.mIsOn =3D 0 } function _TBSetE() { var t=3Dthis if( !t.GetState() ) t.ChgState( gFaceClr,gFaceClr,0,0 ) else t.ChgState( gShadowClr,gHiliteClr,1,1 ) t.mIsOn =3D 1 } function _TBP() { var t=3Dthis if( t.mIsOn ) { if( t.UserAction !=3D null ) t.UserAction() if( !t.GetState ) return if( t.GetState() ) t.SetPressed() else t.SetActive() } } function _TBUI( clr1,clr2,lOffset,tOffset ) { SetBorder( GetObj( this.mObjId ),clr1,clr2 ) Offset( GetObj( this.m_elementsId ),lOffset,tOffset ) } function Offset( obj, top, left ){ obj.style.top=3Dtop; obj.style.left=3Dle= ft } function SetBorder( obj, upperLeft, lowerRight ) { s=3Dobj.style; s.borderStyle =3D "solid" s.borderWidth =3D 1 s.borderLeftColor =3D s.borderTopColor =3D upperLeft s.borderBottomColor=3D s.borderRightColor =3D lowerRight } function GetBtnObj(){ return gBtnArr[window.event.srcElement.id] } function BtnOnOver(){ b=3DGetBtnObj(); if( b !=3D null ) b.SetActive() } function BtnOnDown(){ b=3DGetBtnObj(); if( b !=3D null ) b.SetPressed() } function BtnOnOut(){ b=3DGetBtnObj(); if( b !=3D null ) b.SetInactive() } function BtnOnUp() { b=3DGetBtnObj() if( b !=3D null ) b.Perform() else Upd() } function GetNtsState(){ return parent.gNtsOpen } function GetOtlState(){ return parent.gOtlOpen } function GetOtlTxtState(){ return parent.gOtlTxtExp } function NtsBtnSetFlag( fVal ) { s=3Ddocument.all.item( this.m_flagId ).style s.display=3D"none" if( fVal ) s.display=3D"" else s.display=3D"none" } function _BSetA_Border(){ b =3D gBtnArr[this.mObjId]; if( b !=3D null ) b.S= etActive() } function _BSetI_Border(){ b =3D gBtnArr[this.mObjId]; if( b !=3D null ) b.S= etInactive() } function _BSetP_Border(){ b =3D gBtnArr[this.mObjId]; if( b !=3D null ) b.S= etPressed() } function _BSetA_BorderImg() { b =3D gBtnArr[this.mBorderId] if( b !=3D null && this.mIsOn && !b.GetState() ) { obj=3Dthis.ChgState( gHiliteClr,gShadowClr,2 ) obj.style.posTop=3D0 } } function _BSetI_BorderImg() { b =3D gBtnArr[this.mBorderId] if( b !=3D null && this.mIsOn && !b.GetState() ) { obj=3Dthis.ChgState( gFaceClr,gFaceClr,1 ) obj.style.posTop=3D0 } } var gHiliteClr=3D"THREEDHIGHLIGHT",gShadowClr=3D"THREEDSHADOW",gFaceClr=3D"= THREEDFACE" var gBtnArr =3D new Array() gBtnArr["nb_otl"] =3D new TxtBtn( "nb_otl","nb_otlElem",parent.ToggleOtlPan= e,GetOtlState ) gBtnArr["nb_otlElem"] =3D new TxtBtn( "nb_otl","nb_otlElem",parent.ToggleOt= lPane,GetOtlState ) gBtnArr["nb_nts"] =3D new ImgBtn( "nb_nts","nb_ntsBorder",10,parent.ToggleN= tsPane ) gBtnArr["nb_nts"].SetActive =3D _BSetA_BorderImg; gBtnArr["nb_nts"].SetInactive =3D _BSetI_BorderImg; gBtnArr["nb_ntsBorder"] =3D new TxtBtn( "nb_ntsBorder","nb_ntsElem",parent.= ToggleNtsPane,GetNtsState ) gBtnArr["nb_ntsElem"] =3D new TxtBtn( "nb_ntsBorder","nb_ntsElem",parent.To= ggleNtsPane,GetNtsState ) gBtnArr["nb_prevBorder"] =3D gBtnArr["nb_prev"]=3D new ImgBtn( "nb_prev","n= b_prevBorder",30,parent.GoToPrevSld ) gBtnArr["nb_nextBorder"] =3D gBtnArr["nb_next"]=3D new ImgBtn( "nb_next","n= b_nextBorder",30,parent.GoToNextSld ) gBtnArr["nb_sldshw"]=3D new ImgBtn( "nb_sldshw","nb_sldshwBorder",18,parent= .FullScreen ) gBtnArr["nb_sldshwBorder"] =3D new TxtBtn( "nb_sldshw","nb_sldshwBorder",pa= rent.FullScreen,null ) gBtnArr["nb_sldshwBorder"].SetActive =3D _BSetA_Border; gBtnArr["nb_sldshwBorder"].SetInactive =3D _BSetI_Border; gBtnArr["nb_sldshwText"] =3D new TxtBtn( "nb_sldshw","nb_sldshwText",parent= .FullScreen,null ) gBtnArr["nb_sldshwText"].SetActive =3D _BSetA_Border; gBtnArr["nb_sldshwText"].SetInactive =3D _BSetI_Border; gBtnArr["nb_voice"] =3D gBtnArr["nb_voiceBorder"] =3D new ImgBtn( "nb_voice= ","nb_voiceBorder",18,parent.ToggleVNarration ) gBtnArr["nb_otlTxtBorder"] =3D gBtnArr["nb_otlTxt"]=3D new ImgBtn( "nb_otlT= xt","nb_otlTxtBorder",23,parent.ToggleOtlText ) gBtnArr["nb_ntsBorder"].m_flagId=3D "nb_nts" gBtnArr["nb_ntsBorder"].SetFlag =3D NtsBtnSetFlag gBtnArr["nb_otlTxt"].ChangeIcon=3D GetOtlTxtState var sNext=3D"Další",sPrev=3D"Předchozí",sEnd=3D"Kon= ec prezentace",sFont=3D"Arial",sArrow=3D"Šipka",sFreeform=3D"Voln&yacu= te; tvar",sRect=3D"Obdélník",sOval=3D"Ovál" function ShowMenu() { BuildMenu(); var doc=3DPPTSld.document.body,x=3DPPTSld.event.clientX+doc.scrollLeft,y= =3DPPTSld.event.clientY+doc.scrollTop m =3D PPTSld.document.all.item("ctxtmenu") m.style.pixelLeft=3Dx if( (x+m.scrollWidth > doc.clientWidth)&&(x-m.scrollWidth > 0) ) m.style.pixelLeft=3Dx-m.scrollWidth m.style.pixelTop=3Dy if( (y+m.scrollHeight > doc.clientHeight)&&(y-m.scrollHeight > 0) ) m.style.pixelTop=3Dy-m.scrollHeight m.style.display=3D"" } function _CM() { if( !parent.IsFullScrMode() ) return; if(!PPTSld.event.ctrlKey) { ShowMenu() return false } else HideMenu() } function BuildMenu() { if( PPTSld.document.all.item("ctxtmenu") ) return var mObj=3DCreateItem( PPTSld.document.body ) mObj.id=3D"ctxtmenu" mObj.style.visibility=3D"hidden" var s=3DmObj.style s.position=3D"absolute" s.cursor=3D"default" s.width=3D"120px" SetCMBorder(mObj,"menu","black") var iObj=3DCreateItem( mObj ) SetCMBorder( iObj, "threedhighlight","threedshadow" ) iObj.style.padding=3D2 CreateMenuItem( iObj,sNext,M_GoNextSld,M_True ) CreateMenuItem( iObj,sPrev,M_GoPrevSld,M_HasPrevSld ) CreateSeparator( iObj ) CreateMenuItem( iObj,sEnd,M_End,M_True ) mObj.style.visibility=3D"visible" } function Cancel() { window.event.cancelBubble=3Dtrue; window.event.returnVa= lue=3Dfalse } function Highlight() { ChangeClr("activecaption","threedhighlight") } function Deselect() { ChangeClr("threedface","menutext") } function Perform() { e=3DPPTSld.event.srcElement if( e.type=3D=3D"menuitem" && e.IsActive() ) e.Action() else PPTSld.event.cancelBubble=3Dtrue } function ChangeClr( bg,clr ) { e=3DPPTSld.event.srcElement if( e.type=3D=3D"menuitem" && e.IsActive() ) { e.style.backgroundColor=3Dbg e.style.color=3Dclr } } function M_HasPrevSld() { return( parent.HasPrevSld() ) } function M_GoNextSld() { if( gIsEndShow ) M_End(); else GoToNextSld() } function M_GoPrevSld() { if( gIsEndShow ) { gIsEndShow=3D0; history.back();= PPTSld.event.cancelBubble=3Dtrue; } else GoToPrevSld() } function M_True() { return true } function M_End() { window.close( self ) } function CreateMenuItem( node,text,action,eval ) { var e=3DCreateItem( node ) e.type=3D"menuitem" e.Action=3Daction e.IsActive=3Deval e.innerHTML=3Dtext if( !e.IsActive() ) e.style.color=3D"threedshadow" e.onclick=3DPerform e.onmouseover=3DHighlight e.onmouseout=3DDeselect s=3De.style; s.fontFamily=3DsFont s.fontSize=3D"9pt" s.paddingLeft=3D2 } function CreateSeparator( node ) { var sObj=3DCreateItem( node ) SetCMBorder(sObj,"menu","menu") var s=3DsObj.style s.borderTopColor=3D"threedshadow" s.borderBottomColor=3D"threedhighlight" s.height=3D1 s.fontSize=3D"0px" } function CreateItem( node ) { var elem=3DPPTSld.document.createElement("DIV") node.insertBefore( elem ) return elem } function SetCMBorder( o,ltClr,rbClr ) { var s=3Do.style s.backgroundColor=3D"menu" s.borderStyle=3D"solid" s.borderWidth=3D1 s.borderColor=3DltClr+" "+rbClr+" "+rbClr+" "+ltClr } ------=_NextPart_01CA626C.98BFB110 Content-Location: file:///C:/A37AC9E1/kontrola_soubory/fullscreen.htm Content-Transfer-Encoding: quoted-printable Content-Type: text/html; 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charset="us-ascii" Prezentace aplikace PowerPoint ------=_NextPart_01CA626C.98BFB110 Content-Location: file:///C:/A37AC9E1/kontrola_soubory/outline.htm Content-Transfer-Encoding: quoted-printable Content-Type: text/html; charset="us-ascii"

Po= známky

Prezentac= e

Osnova

1

ANALYTICKÉ ZNAKY METOD
KONTROLA KVALITY V KLINICKÉ LABORATOŘI

Martin Beránek

UKBD FN HK

Přednáška pro FaF

2

Analytické= ; znaky metody jsou testovány v rámci validačního proce= su

3

Analytické= ; znaky metod

Diagnostická citlivost a specifičnost

Analytická citlivost

Mez detekce a mez stanovitelnosti

Analytická specifičnost

Výtěžnost

Linearita

Pracovní rozsah

Přesnost a správnost (=3D spolehlivost, validita)

Proveditelnost

4

"nejmen#= 3;í množství analytu"

nejmenší množství analytu, které lze ještě s určitou spolehlivostí stanovit

nejmenší množství analytu, které zp&#= 367;sobí měřitelnou výchylku (signál na detektoru)

       = ;            &n= bsp;            = ;      ΔD

       = ;          tgα =3D -----------   …. směrnice kalibrační funkce

       = ;            &n= bsp;            = ;      Δc

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Mez detekce (lim= it of detection- LD)

nejmenší množství analytu, které může být ve vzorku kvalitativně dokázáno

       = ;             L_D =3D blank + 3*s_bl_{=
;            &n=
bsp;
SD =3D s}

hodnota vyjadřuje schopnost analytického systému = rozlišit přístrojový šum od analytického signálu

číselně definuje přístrojový šum

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Mez stanovitelno= sti (Lq)

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Analytické= ; metody o vyšší analytické citlivosti vykazují nižší meze detekce a stanovitelnosti

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Analytick&aacut= e; specifičnost metody

Vlastnost metody, která vyjadřuje, jak stanovení analytu ovlivňuje (ruší, interferuje) přítomnost dalších látek endogenního či exogenního původu ve vyšetřovaném biologickém materiálu
Analytická interference látek

Analytická interference samotné matrice

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Analytick&aacut= e; specifičnost

Analytická interference způsobuje systematickou chybu měření

Hb, lipidy, bilirubin, antikoagulancia při bch analýze s= ér

křížové ICH reakce (aldosteron vs kortison)<= /li>
kyselina askorbová při chemické analýze moči

léky podobné struktury při toxikologické analýze

Analytickou specifičnost zjistíme porovnání= ;m s referenční metodou (hodnotíme % odchylku s rostoucím přídavkem interferentu vůči povoleným mezím)

Interference klinické (diuretika, indukce EE P450,…)

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Analytick&aacut= e; interference bilirubinu
(od koncentrace 0,5 g/l)

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Výpo= 9;et výtěžnosti (recovery)

       = ; Naměřená hodnota cholesterolu

v séru (hodnota cílová):       = ;    5,86 mmol/l

ve frakci LDL + HDL LP:       4,25 mmol/l

ve frakci CHM + VLDL LP:   1,18 mmol/l

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Linearita metod= y

Rozsah kalibrační funkce s lineárním průběhem,

Jde o určitou část (úsek) každé = funkce!

Výsledek se kvantifikuje směrnicí přímky    y =3D B*x + A

Ověření linearity

       = ; sadou RM v příslušné koncentrační řadě

       = ;    - vytvořením ředící řady z nativního materiálu vhodným

       = ;      rozpouštědlem (negativní vzorek, fyziologický

       = ;      roztok, 0. kalibrátor,…)

       = ;     Správnost ředění !!

       = ;     Shoda průběhů kalibrátor vs biologický materiál!!

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Pracovní= rozsah

    U line&aa= cute;rních metod, resp. při měření v lineárn&iacut= e; oblasti dána dolní a horní mez linearity (záruka spolehlivosti stanovení analytu)

   U nelineá= ;rních metod o pracovním rozsahu metody rozhoduje rozsah hodnot komerčních kalibrátorů

       při měření hodnocení: „pod= /nad pracovním rozsahem“

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"Absolutn&= iacute; (definitivní"

      Absolutní (definitivní) metoda

        gravimetrie, odměrné titrace, coulometrie

       = ;            Bias 0%   C= Va 0%

       = ;            &n= bsp; Referenční metoda

důkladně ustanovený, prostudovaný, definovaný

      certifikovaný a validovaný měřící postup

    HPLC-MS, sekvenování DNA, HPCE, AAS

       = ;            Bias 0%    CVa ~ 1%

       = ;            &n= bsp;    Standardizovaná

       = ;        rutinní metoda

      Bias dle návaznosti   CVa 1-10%

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     Kalibrá= tory v klinické biochemii

Primární kalibrátor – analyt v rozpouštědle

       = ;            &n= bsp;            = ;   - výhoda: čistota, stabilita

       = ;            &n= bsp;            = ;   - nevýhoda: nezohledňují matrici

       = ;      použití pro: kalibrace referenčních m= etod

Sekundární kalibrátor – analyt v matricovém základu

      (komutabilní s biologickými materiály) s hodnotami

      určenými referenčními metodami či odvozené z

      certifikovaných RM;&n= bsp; pro rutinní kalibrace (a EHK)

Multikalibrátor – sekundární kalibrátor obsahující více

       = ;            &n= bsp;            rozpuštěných analytů

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Komutabilita

=3D shoda vlastností referenčního materiálu (kalibrátoru, kontroly) s reálnými vzorky v průběhu měření

… aby materiál byl co nejvíc nativní

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Kdy kalibrovat a rekalibrovat?

Nový stroj, nová aplikace, nové analytick&eacut= e; podmínky

Požadavek výrobce (četnost dle stability činid= el, nové šarže)

Po opravě a údržbě stroje

Velká systematická chyba měření (EHK)=

22

Kontrola kvalit= y
(práce v laboratoři)

Vnitřní kontrola kvality (VKK, IQC)

    Informuje= o stabilitě analytického systému

    Zá= kladem dlouhodobé hodnocení CVa (%)

Externí kontrola kvality (EHK, EQA)

   Informuje o por= ovnatelnosti laboratoře, tedy o systematické chybě

   Základem hodnocení bias (a výpočet z skore)

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Westgardova kontrolní pravidla VKK        1_3s, 2_2s, R_3s, 10x

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Hodnocení úspěšnosti v EHK

Cílová hodnota (TV) ± 2 SD

(interval spolehlivosti 95%)

Toleranční rozpětí TE_max (HL R= 11; DL)

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"Cí= lová hodnota RM získaná změřením..."

Cílová hodnota RM získaná změřením analytu

       = ;       referenční metodou (např. glukosa)

Cílová hodnota RM získaná v několika expertních

      laboratořích metodou doporučenou IFCC (enzym= y)

Cílová hodnota získaná jako prům= 83;r účastníků EHK

       = ;            &n= bsp;     používajících tutéž analytickou metodu

       = ;        (hormony metodou RIA firmy Amersham)

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Co patří do znaků proveditelnosti ?

Čas odezvy mezi příjmem materiálu a výdejem výsledku po celých 24 hodin

Přímé a nepřímé náklady= a platba ZP

Analytické, technické, ekonomické a logistické zabezpečení laboratoře

       = ;       viz laboratorní dokumentace

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Analytick&eacut= e; cíle

Rozvoj technologie – přesnější analýzy

   (↓ CV)
Nové referenční metody a komutabilnější materiály – vyšší návaznost (↓ bias)

Nové statistické metody a výpočetní technika

Snižování nejistoty v pre- a post-analyt. F&aacut= e;zi

Snižování nejistoty vs. cena vyšetření

       = ;            &n= bsp;      … existují smysluplné hranice ?

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Základem standardní laboratorní práce je především:

Pečlivě číst příbalové informace výrobce

Provádět kontrolu kvality (KK, QC)

Vést záznamy o technickém stavu přístrojů

Vést pečlivou laboratorní dokumentaci

= = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = =
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1	ANALYTICKÉ ZNAKY METOD KONTROLA KVALITY V KLINICKÉ LABORATOŘI Martin Beránek UKBD FN HK Přednáška pro FaF
2	Analytické= ; znaky metody jsou testovány v rámci validačního proce= su
3	Analytické= ; znaky metod Diagnostická citlivost a specifičnost Analytická citlivost Mez detekce a mez stanovitelnosti Analytická specifičnost Výtěžnost Linearita Pracovní rozsah Přesnost a správnost (=3D spolehlivost, validita) Proveditelnost
4	"nejmen#= 3;í množství analytu" nejmenší množství analytu, které lze ještě s určitou spolehlivostí stanovit nejmenší množství analytu, které zp&#= 367;sobí měřitelnou výchylku (signál na detektoru) = ; &n= bsp; = ; ΔD = ; tgα =3D ----------- …. směrnice kalibrační funkce = ; &n= bsp; = ; Δc
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6	Mez detekce (lim= it of detection- LD) nejmenší množství analytu, které může být ve vzorku kvalitativně dokázáno = ; L_D =3D blank + 3*s_bl_{= ; &n= bsp; SD =3D s} hodnota vyjadřuje schopnost analytického systému = rozlišit přístrojový šum od analytického signálu číselně definuje přístrojový šum
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8	Mez stanovitelno= sti (Lq)
9	Analytické= ; metody o vyšší analytické citlivosti vykazují nižší meze detekce a stanovitelnosti
10	Analytick&aacut= e; specifičnost metody Vlastnost metody, která vyjadřuje, jak stanovení analytu ovlivňuje (ruší, interferuje) přítomnost dalších látek endogenního či exogenního původu ve vyšetřovaném biologickém materiálu Analytická interference látek Analytická interference samotné matrice
11	Analytick&aacut= e; specifičnost Analytická interference způsobuje systematickou chybu měření Hb, lipidy, bilirubin, antikoagulancia při bch analýze s= ér křížové ICH reakce (aldosteron vs kortison)<= /li> kyselina askorbová při chemické analýze moči léky podobné struktury při toxikologické analýze Analytickou specifičnost zjistíme porovnání= ;m s referenční metodou (hodnotíme % odchylku s rostoucím přídavkem interferentu vůči povoleným mezím) Interference klinické (diuretika, indukce EE P450,…)
12	Analytick&aacut= e; interference bilirubinu (od koncentrace 0,5 g/l)
13	Výpo= 9;et výtěžnosti (recovery) = ; Naměřená hodnota cholesterolu v séru (hodnota cílová): = ; 5,86 mmol/l ve frakci LDL + HDL LP: 4,25 mmol/l ve frakci CHM + VLDL LP: 1,18 mmol/l
14	Linearita metod= y Rozsah kalibrační funkce s lineárním průběhem, Jde o určitou část (úsek) každé = funkce! Výsledek se kvantifikuje směrnicí přímky y =3D B*x + A Ověření linearity = ; sadou RM v příslušné koncentrační řadě = ; - vytvořením ředící řady z nativního materiálu vhodným = ; rozpouštědlem (negativní vzorek, fyziologický = ; roztok, 0. kalibrátor,…) = ; Správnost ředění !! = ; Shoda průběhů kalibrátor vs biologický materiál!!
15	Pracovní= rozsah U line&aa= cute;rních metod, resp. při měření v lineárn&iacut= e; oblasti dána dolní a horní mez linearity (záruka spolehlivosti stanovení analytu) U nelineá= ;rních metod o pracovním rozsahu metody rozhoduje rozsah hodnot komerčních kalibrátorů při měření hodnocení: „pod= /nad pracovním rozsahem“
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17	"Absolutn&= iacute; (definitivní" Absolutní (definitivní) metoda gravimetrie, odměrné titrace, coulometrie = ; Bias 0% C= Va 0% = ; &n= bsp; Referenční metoda důkladně ustanovený, prostudovaný, definovaný certifikovaný a validovaný měřící postup HPLC-MS, sekvenování DNA, HPCE, AAS = ; Bias 0% CVa ~ 1% = ; &n= bsp; Standardizovaná = ; rutinní metoda Bias dle návaznosti CVa 1-10%
18	Kalibrá= tory v klinické biochemii Primární kalibrátor – analyt v rozpouštědle = ; &n= bsp; = ; - výhoda: čistota, stabilita = ; &n= bsp; = ; - nevýhoda: nezohledňují matrici = ; použití pro: kalibrace referenčních m= etod Sekundární kalibrátor – analyt v matricovém základu (komutabilní s biologickými materiály) s hodnotami určenými referenčními metodami či odvozené z certifikovaných RM;&n= bsp; pro rutinní kalibrace (a EHK) Multikalibrátor – sekundární kalibrátor obsahující více = ; &n= bsp; rozpuštěných analytů
19	Komutabilita =3D shoda vlastností referenčního materiálu (kalibrátoru, kontroly) s reálnými vzorky v průběhu měření … aby materiál byl co nejvíc nativní
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21	Kdy kalibrovat a rekalibrovat? Nový stroj, nová aplikace, nové analytick&eacut= e; podmínky Požadavek výrobce (četnost dle stability činid= el, nové šarže) Po opravě a údržbě stroje Velká systematická chyba měření (EHK)=
22	Kontrola kvalit= y (práce v laboratoři) Vnitřní kontrola kvality (VKK, IQC) Informuje= o stabilitě analytického systému Zá= kladem dlouhodobé hodnocení CVa (%) Externí kontrola kvality (EHK, EQA) Informuje o por= ovnatelnosti laboratoře, tedy o systematické chybě Základem hodnocení bias (a výpočet z skore)
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25	Westgardova kontrolní pravidla VKK 1_3s, 2_2s, R_3s, 10x
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28	Hodnocení úspěšnosti v EHK Cílová hodnota (TV) ± 2 SD (interval spolehlivosti 95%) Toleranční rozpětí TE_max (HL R= 11; DL)
29	"Cí= lová hodnota RM získaná změřením..." Cílová hodnota RM získaná změřením analytu = ; referenční metodou (např. glukosa) Cílová hodnota RM získaná v několika expertních laboratořích metodou doporučenou IFCC (enzym= y) Cílová hodnota získaná jako prům= 83;r účastníků EHK = ; &n= bsp; používajících tutéž analytickou metodu = ; (hormony metodou RIA firmy Amersham)
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33	Co patří do znaků proveditelnosti ? Čas odezvy mezi příjmem materiálu a výdejem výsledku po celých 24 hodin Přímé a nepřímé náklady= a platba ZP Analytické, technické, ekonomické a logistické zabezpečení laboratoře = ; viz laboratorní dokumentace
34	Analytick&eacut= e; cíle Rozvoj technologie – přesnější analýzy (↓ CV) Nové referenční metody a komutabilnější materiály – vyšší návaznost (↓ bias) Nové statistické metody a výpočetní technika Snižování nejistoty v pre- a post-analyt. F&aacut= e;zi Snižování nejistoty vs. cena vyšetření = ; &n= bsp; … existují smysluplné hranice ?
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36	Základem standardní laboratorní práce je především: Pečlivě číst příbalové informace výrobce Provádět kontrolu kvality (KK, QC) Vést záznamy o technickém stavu přístrojů Vést pečlivou laboratorní dokumentaci