KR18 IA2Ab ELISA x1a - kronus.com
2
IA-2 Autoantibody (IA-2Ab) The KRONUS IA-2 Autoantibody (IA-2Ab) ELISA Kit is for the quantitative determination of autoantibodies to Islet Antigen-2 (IA-2) in human serum and may be useful as an aid in the diagnosis of Type 1 diabetes mellitus (autoimmune mediated diabetes). The IA-2Ab ELISA Kit is not to be used alone and is to be used in conjunction with other clinical and laboratory findings. The KRONUS IA-2Ab ELISA Kit depends on the ability of IA-2 autoantibodies to act divalently, forming a bridge between IA-2 coated on ELISA plate wells and liquid phase IA-2-biotin. The resulting antigen-antibody- antigen complexes are then quantitated by the addition of streptavidin peroxidase (SA-POD) and tetramethylbenzidine (TMB) to produce a colorogenic reaction. Stop solution is added to halt the reaction and absorbance is read using an ELISA plate reader. The absorbance of each well is directly proportional to the amount of antibody present. IA-2Ab levels are derived from a standard curve, traceable to the WHO reference preparation NIBSC 97/550 and expressed in U/mL. ASSAY CHARACTERISTICS Quantitative Calibration: 5 Calibrators, 0.75 – 350 U/mL NIBSC 97/550 Controls (Included): 1 Positive, 1 Negative TOTAL RUNNING TIME 17 Hours and 40 Minutes SPECIMEN MATRIX Human Serum REFERENCE RANGE < 7.5 U/mL: Negative ≥ 7.5 U/mL: Positive PRECISION Intra-Assay Inter-Assay Dose (U/mL) % CV Dose (U/mL) % CV 7.3 6.69 7.3 9.69 28.1 5.05 28.1 6.46 82.9 2.89 82.9 4.92 PATIENT NUMBER OF PATIENTS GROUP POSITIVE FOR IA-2 Ab % Type 1 Diabetes 139/239 58.2 LADA 10/45 22.2 Type 2 Diabetes 2/57 3.5 Rheumatoid Arthritis 1/20 5.0 Hashimoto’s thyroiditis 1/20 5.0 Graves’ Disease 0/19 0.0 Systemic Lupus Erythematosus 0/14 0.0 Metabolic Syndrome 1/35 2.8 Addison’s Disease 3/27 11.1 Celiac Disease 0/50 0.0 Healthy Blood Donors 14/718 1.9 CLINICAL SENSITIVITY & SPECIFICITY Sensitivity 58.2% Specificity 97.0% ELISA For In Vitro Diagnostic Use. Rx Only. ISLET CELL AUTOIMMUNITY ORDERING INFORMATION KR7755 — 96 Well Kit 800.4.KRONUS (800.457.6687) | 208.377.4800 | www.kronus.com | [email protected]
Transcript of KR18 IA2Ab ELISA x1a - kronus.com
IA-2 Autoantibody (IA-2Ab)
The KRONUS IA-2 Autoantibody (IA-2Ab) ELISA Kit is for the quantitative determination of autoantibodies to Islet Antigen-2 (IA-2) in human serum and may be useful as an aid in the diagnosis of Type 1 diabetes mellitus (autoimmune mediated diabetes). The IA-2Ab ELISA Kit is not to be used alone and is to be used in conjunction with other clinical and laboratory findings.
The KRONUS IA-2Ab ELISA Kit depends on the ability of IA-2 autoantibodies to act divalently, forming a bridge between IA-2 coated on ELISA plate wells and liquid phase IA-2-biotin. The resulting antigen-antibody-antigen complexes are then quantitated by the addition of streptavidin peroxidase (SA-POD) and tetramethylbenzidine (TMB) to produce a colorogenic reaction. Stop solution is added to halt the reaction and absorbance is read using an ELISA plate reader. The absorbance of each well is directly proportional to the amount of antibody present. IA-2Ab levels are derived from a standard curve, traceable to the WHO reference preparation NIBSC 97/550 and expressed in U/mL.
ASSAY CHARACTERISTICS
Quantitative Calibration: 5 Calibrators, 0.75 – 350 U/mL NIBSC 97/550
Controls (Included): 1 Positive, 1 Negative
TOTAL RUNNING TIME
17 Hours and 40 Minutes
SPECIMEN MATRIX
Human Serum
REFERENCE RANGE
< 7.5 U/mL: Negative≥ 7.5 U/mL: Positive
PRECISION
Intra-Assay Inter-Assay Dose (U/mL) % CV Dose (U/mL) % CV 7.3 6.69 7.3 9.69 28.1 5.05 28.1 6.46 82.9 2.89 82.9 4.92
PATIENT NUMBER OF PATIENTSGROUP POSITIVE FOR IA-2 Ab %Type 1 Diabetes 139/239 58.2LADA 10/45 22.2Type 2 Diabetes 2/57 3.5Rheumatoid Arthritis 1/20 5.0Hashimoto’s thyroiditis 1/20 5.0Graves’ Disease 0/19 0.0Systemic Lupus
Erythematosus 0/14 0.0Metabolic Syndrome 1/35 2.8Addison’s Disease 3/27 11.1Celiac Disease 0/50 0.0Healthy Blood Donors 14/718 1.9
CLINICAL SENSITIVITY & SPECIFICITY
Sensitivity 58.2%Specificity 97.0%
ELISA
For In Vitro Diagnostic Use. Rx Only.
ISLE
T CE
LL A
UTOI
MM
UNIT
Y
ORDERING INFORMATION
KR7755 — 96 Well Kit
800.4.KRONUS (800.457.6687) | 208.377.4800 | www.kronus.com | [email protected]
KR18_IA2Ab_ELISA.indd 1 5/18/18 12:32 PM
…Your Source for Sensitive Autoimmune Diagnostics
ASSAY PROCEDURE
Sample Volume: 50 µL per Well
Calibrators, Controls and Samples into Coated Wells: 50 µL
Reaction Enhancer: 25 µL
Overnight Incubation at 2 – 8° C
Wash Wells 3 Times
IA-2-Biotin: 100 µL
1 Hour Incubation at 2 – 8° C
Wash Wells 3 Times
SA-POD: 100 µL
20 Minute Incubation with Shaking at Room Temperature
Wash Wells 3 Times
TMB Substrate: 100 µL
20 Minute Incubation at Room Temperature
Stop Solution: 100 µL
Read Absorbance at 450nm
Total Assay Time is Approximately 17 Hours and 40 Minutes
Intended for informational purposes only. Please see Product Direction Insert for complete details, instructions, limitations, etc. 05/18Manufactured Under License to US Patents 8,129,132 B2 and 9,435,797 B2.
1. Borg H, Fernlund P & Sundkvist G, ‘Measurement of antibodies against glutamic acid decarboxylase 65 (GADA): two new 125I assays compared with [35S] GAD 65-ligand binding assay’, Clinical Chemistry, 1997, 43(5); 779-785.
2. Pardini VC, Mourão DM, Nascimento PD et al, ‘Frequency of islet cell autoantibodies (IA-2 and GAD) in young Brazilian type 1 diabetes patients’, Brazilian Journal of Medical and Biological Research, 1999, 32: 1195-1198.
3. Winter WE, Harris N & Schatz D, ‘Type 1 diabetes islet autoantibody markers’, Diabetes Technology & Therapeutics, 2002, 4(6); 817-839.
4. S. Chen et al. Sensitive non isotopic assays for autoantibodies to IA-2 and to a combination of both IA-2 and GAD65., Clinica Chimica Acta 2005 357:74-83.
5. E. Nilsson et al. Calcium addition to EDTA plasma eliminates falsely positive results in the RSR GADAb ELISAClinica Chimica Acta 388 (2008) 130-134.
6. K. Rahmati et al. A Comparison of Serum and EDTA Plasma in the Measurement of Glutamic Acid Decarboxylase Autoantibodies (GADA) and Autoantibodies to Islet Antigen-2 (IA-2A) Using the RSR Radioimmunoassay (RIA) and Enzyme Linked Immunosorbent Assay (ELISA) Kits., Clin. Lab. 2008 54:227-235.
7. C. Törn et al. Diabetes Antibody Standardization Program: evaluation of assays for autoantibodies to glutamic acid decarboxylase and islet antigen- 2. Diabetologia 2008 51:846-852
REFERENCES
KR18_IA2Ab_ELISA.indd 2 5/18/18 12:32 PM
![EA763AD-54A...(JEMI 426)] (JEM1426)J 3 ELISA ) ELISA ) ELISA ) ELISA ELISA ) ELISA ) JIS Ll 902 JIS Z291 1 JIS Z2801 JIS Z2801 JIS Z2801 JIS Z2801 JIS Z2801 JIS 2911 JIS Z2801 JIS](https://static.fdocuments.net/doc/165x107/5e93b98e09aa5216734c1831/ea763ad-54a-jemi-426-jem1426j-3-elisa-elisa-elisa-elisa-elisa-elisa.jpg)
