CLIA YUNIATI VALENTINA Immunoserology Div – Clinical Pathology Departement FK Unpad - RSHS
PENDAHULUAN Chemiluminescence adalah emisi atau pancaran cahaya oleh produk yang distimulus oleh suatu reaksi kimia atau suatu kompleks cahaya. Cahaya yang dihasilkan menunjukkan jumlah analit yang diperiksa. Chemiluminescence memiliki prinsip dasar kerja ELISA, namun menggunakan Luxogenic Substrate menggantikan Chromogenic Substrate yang biasa dipakai ELISA. Satuan energy cahaya yang dihasilkan Relative Light Unit (RLU)
The suitable SUBSTRATES for luminescent immunoassays include: The widely used ENZYMES for Luminescent Immunoassay are AP (Alkaline Phospatase) , HRP (Horse radish Peroxidase) and Pyruvate Kinase The suitable SUBSTRATES for luminescent immunoassays include: 1. Luminol 2. Acridine Esters 3. Isoluminol 4. Luciferin 5. AMPPD
CLIA butuh trigger ATAU enchancer berupa substansi asam basa seperti OH-/ H202 memicu reaksi oksidasi yang akan menghasilkan warna LUMINOL + H202 OXIDIZED LUMINOL + +H20 OXIDIIZED LUMINOL + HV (LIGHT)
CHEMILUMINESCENT IMMUNOASSAYS Can attach label to antigen or antibody Heterogenous assays use competitive and sandwich assay Competitive assays used to measure smaller analytes Sandwich assays are used to measure larger analytes
SANDWICH CLIA
COMPETITIVE LIGHTS
ADVIA XPT CENTAUR XPT CENTAUR Direct Chemiluminescence Reaction. Directly measure the lights energy without use of added steps or amplifying molecules. Assays use Acridinium Ester (AE)as the chemiluminescence label, because AE does not require the addition of catalyst or substrate XPT CENTAUR uses the dimethyl form of AE because its stability allows long reagent shelf life
HENRY CLINICAL DIAGNOSIS 23rd ED
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