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Samenvatting Klinische chemie - MLT03 - Werken In Een Hemato-klinisch Lab (MLT03)
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Volledige samenvatting van het deel Klinische chemie met bijhorende afbeeldingen.
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1 Overzicht van toestellen, analyse- en detectiemethodologieVerschillende afdelingen binnen het labo
▪ Microbiologie
▪ Hematologie
▪ Pathologie
▪ Klinische chemie (schade/functies organen, stofwisselingen)
o Stalen: urine, bloed, lichaamsvochten
o Waarom?: diagnose stellen, opvolging behandeling, preventie
Verschillende parameters:
▪ Cellen (bloed) → hematologie
▪ Biochemische parameters: eiwitten, enzymen
▪ Chemische parameters: ionen, vitamines, mineralen
▪ Karakteristieken: stolling, GFR, osmolaliteit
Verschillende analyses in een klinisch labo
▪ Biochemie:
o Elektrochemie → elektroden (spanning/stroom), ISE (Na+, K+, …)
o Enzymatische assays → spectrofotometrie
o Chemische assays → spectrofotometrie
▪ Immunochemie → nefelometrie, turbidimetrie → lichtverstrooiing
→ (sandwich-, competitieve) ELISA → fluorochroom, radio-isotoop
gelabeld enzym (AF), elektrochemiluminiscentie
Enzymatische assays:
1.1.1 Verloop enzymreactie
Enzym + substraat → enzymsubstraat complex → enzym + producten
Verloop reactie volgen: toename [gevormde product] of afname [verbruikte substraat]
Product of substraat meetbaar bij bepaalde golflengte → extincties uitzetten tegen tijd →
lineair verband
Na enige tijd: curve vlakt af door verlaging substraatconcentratie, inactivatie enzymes, …
Invloedsfactoren:
▪ T ↑ → enzymreactie ↑
▪ pH: optimaal voor elk enzym verschillend → steeds in gebufferde oplossingen
werken
▪ Hoeveelheid substraat: lage conc = rechtevenredige reactiesnelheid, hoge conc =
maximale snelheid (neemt niet meer verder toe)
▪ Enzymconcentratie ↑ → enzymreactie ↑
▪ Enzym-cofactoren, vb NADH omgezet in NAD+, nodig voor reacties Mg2+, Mn2+,
Fe2+, Ca2+ en Zn2+
1.1.2 Bepalen enzymactiviteit
Eenheid: 1 µmol/min (product gevormd of substraat omgezet)
Juiste omstandigheden! (T, pH via buffer, substraat conc, aanwezigheid cofactore,
afwezigheid remmers)
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