Thyroxine T4 Neonatal (NNT4) BioAssay(TM) ELISA Kit

The Quantitative Determination of Total Thyroxine Concentration in Human (Neonates) whole blood by a Microwell Enzyme Immunoassay \n\nDetermination of hypothyroidism within the first few days of birth has been recognized as the single most important diagnostic test in neonates by the American Thyroid Association The need for its early detection and treatment has resulted in the establishment of screening centers by federal and state health departments, A program of early screening of neonates for congenital hypothyroidism was started in Quebec, Canada in the early seventies. They used dry blood spots on filter paper as the sampling device. Very soon the program was followed by other major public health institutions in Canada and the US. By 1978, almost one million infants had \nbeen screened and an incidence rate of congenital hypothyroidism was established to be approximately 1 in 7000 births. Congenital hypothyroidism is probably the single most common preventable cause of mental retardation. Diagnosis and treatment of congenital hypothyroidism within the first 1-2 months after birth appears to be necessary in order to prevent severe mental retardation. This microplate enzyme immunoassay methodology provides the technician with optimum sensitivity while requiring few technical manipulations. In this method, calibrators, patient specimen, or controls, all made and dried in whole blood are first added to a microplate well. A buffer containing essential ingredients to isolate T4 from blood proteins is added. The blood from the filter paper dots is allowed to elute in the buffer. In the process, T4 (Thyroxine) dissociates from the serum (blood) proteins and binds to the antibody that is immobilized on the inside of the microwells. Excess blood is removed using a wash step. Enzyme-T4 conjugate \nis added. The enzyme labeled T4 binds to the sites on the antibody left available by the native T4 that came from the sample. After the completion of the required incubation period, excess enzyme conjugate is removed using a wash step. The activity of the enzyme present on the surface of the well is quantitated by reaction with a suitable substrate to produce color. The employment of several references, made in whole blood, of known thyroxine concentration permits construction of a dose response curve (DRC-graph) of activity and con-centration. An unknown specimen's activity can be extrapolated from the DRC. \n\nDetection Range: 0-25ug/dL \n\nSample: 50ul \n\nSpecificity: ~97% \n\nSensitivity: 0.05ug/dL \n\nTotal Time: ~75min \n \nKit Components: \nEach kit contains the following components in sufficient quantities to perform the number of tests indicated. All reactive reagents contain 0.1% sodium azide as a preservative. \n1. Microtiter Plate\n2. Conjugate\n3. Calibrator\n4. Positive Control\n5. Negative Control\n6. TMB\n7. Wash Buffer, 10X Diluted wash buffer to 1X\n8. Sample Diluent\n9. Stop Solution \n\nStorage and Stability:\nStore all components at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

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