ISOLATION OF THE EXTRACTABLE NUCLEAR ANTIGENS (ENA) TO DESIGN ENA-DID KITS AND DETERMINE ITS VALIDITY BY EVALUATING-OF SYSTEMIC RHEUMATOID DISEASE

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VARASTEH A.; SAGHAFI M.; BARADARAN H.; MAHMOUDI M.; REZAEI A.; TALAEI N.; RAHIMZADEH ABDI F.

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Journal: Iranian Journal of Basic Medical Sciences Year:2001 | Volume:3 | Issue:3-4 (8) Page(s): 155-164

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Title

ISOLATION OF THE EXTRACTABLE NUCLEAR ANTIGENS (ENA) TO DESIGN ENA-DID KITS AND DETERMINE ITS VALIDITY BY EVALUATING-OF SYSTEMIC RHEUMATOID DISEASE

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Keywords

EXTRACTABLE NUCLEAR ANTIGENS (ENA)Q4

DID KITQ4

Abstract

Determination of autoantibodies to nuclear antigens is important in diagnosis of systemic rheumatoid diseases. Among these autoantibodies, antibodies to extractable nuclear antigens (Sm, nRNP, SS/A and SS/B) play a more specific role in differentiated diagnosis of these diseases. In the current study we have extracted the above mentioned antigens from calf thymus and human spleen. Calf thymus was used for extraction' of Sm, nRNP and SS/B. Unclear extract was first prepared using ultracentrifugation (35,000 rpm 2 hrs); then proteins were precipitated by 30% to 60% ammonium sulfate. Sm/nRNP complex was treated by RNase for isolation of Sm. In the next stage isolated protein precipitate was treated by 60% to 80% ammonium sulfate to prepare SS/B. SS/A was prepared from human spleen cytoplasm extracts. Human splenocytes were centrifuged at 40,000 rpm for 3 hrs. Then protein precipitate was isolated using 30%to 60% ammonium sulfate, which contained SS/A. The presence of extracted antigens was confirmed by western blotting. The extracted antigens were used in designing and optimization of relevant DID diagnostic kits. These kits had 100% precision in inter and intra assays. In SLE patients the sensitivity and specificity of anti-Sm- DID KIT were 35% and 95% respectively. In MCTD patients sensivity and specificity of anti-nRNP. DID KIT were 100% and 84% respectively. In SS patients the sensitivity and specificity of anti-SS/A-DID kit were 55% and 80% respectively, while the sensitivity and specificity of anti-SS!B-DID kit were 44.8% and 85% respectively.

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APA: Copy

VARASTEH, A., SAGHAFI, M., BARADARAN, H., MAHMOUDI, M., REZAEI, A., TALAEI, N., & RAHIMZADEH ABDI, F.. (2001). ISOLATION OF THE EXTRACTABLE NUCLEAR ANTIGENS (ENA) TO DESIGN ENA-DID KITS AND DETERMINE ITS VALIDITY BY EVALUATING-OF SYSTEMIC RHEUMATOID DISEASE. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, 3(3-4 (8)), 155-164. SID. https://sid.ir/paper/52528/en

Vancouver: Copy

VARASTEH A., SAGHAFI M., BARADARAN H., MAHMOUDI M., REZAEI A., TALAEI N., RAHIMZADEH ABDI F.. ISOLATION OF THE EXTRACTABLE NUCLEAR ANTIGENS (ENA) TO DESIGN ENA-DID KITS AND DETERMINE ITS VALIDITY BY EVALUATING-OF SYSTEMIC RHEUMATOID DISEASE. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES[Internet]. 2001;3(3-4 (8)):155-164. Available from: https://sid.ir/paper/52528/en

IEEE: Copy

A. VARASTEH, M. SAGHAFI, H. BARADARAN, M. MAHMOUDI, A. REZAEI, N. TALAEI, and F. RAHIMZADEH ABDI, “ISOLATION OF THE EXTRACTABLE NUCLEAR ANTIGENS (ENA) TO DESIGN ENA-DID KITS AND DETERMINE ITS VALIDITY BY EVALUATING-OF SYSTEMIC RHEUMATOID DISEASE,” IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, vol. 3, no. 3-4 (8), pp. 155–164, 2001, [Online]. Available: https://sid.ir/paper/52528/en

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