دورية أكاديمية
Quantitative Evaluation of the Impact of Ethylenediaminetetraacetic Acid Pretreatment on Single-Antigen Bead Assay
| العنوان: | Quantitative Evaluation of the Impact of Ethylenediaminetetraacetic Acid Pretreatment on Single-Antigen Bead Assay |
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| المؤلفون: | Chang Liu, MD, PhD, Sue Pang, BA, Donna Phelan, BA, Daniel C. Brennan, MD, Thalachallour Mohanakumar, PhD |
| المصدر: | Transplantation Direct, Vol 3, Iss 8, p e194 (2017) |
| بيانات النشر: | Wolters Kluwer, 2017. |
| سنة النشر: | 2017 |
| المجموعة: | LCC:Surgery |
| مصطلحات موضوعية: | Surgery, RD1-811 |
| الوصف: | Background. Ethylenediaminetetraacetic acid (EDTA) pretreatment has been shown to overcome complement interference in the single-antigen bead (SAB) assay. However, a quantitative evaluation of its impact on the assay for preemptive application to diverse clinical samples is still lacking. Methods. Serum samples from 95 renal transplant candidates were tested with and without EDTA-pretreatment in parallel. Changes in mean fluorescence intensity (MFI) values were analyzed to determine the impact of EDTA-pretreatment and the characteristics of complement interference. Results. MFI values from EDTA-treated and untreated sera showed good correlations (r = 0.99) and were linear after excluding outliers (slopes, 1; intercepts, −63.7 and −24.2 for class I and II, respectively). Using an assay cutoff of 2000 MFI, positive/negative assignments were concordant for 99% of the 9215 class I beads and 9025 class II beads tested. As defined by an MFI increment above 4000 after EDTA pretreatment, complement interference affected 172 class I beads in 12 samples (12.6%) and 60 class II beads in 7 samples (7.4%), and the findings were supported in 83% and 86% of these samples by dilution studies. In a case study, EDTA pretreatment prevented falsely low MFI values and facilitated the interpretation of titration curves. Finally, EDTA pretreatment reduced the coefficient of variance (CV) by 2.1% and 2.4% for class I and II beads respectively (P < 0.0001). Conclusions. It is safe to preemptively treat all clinical samples with EDTA before SAB assay to prevent false negative results or falsely low MFI values. EDTA pretreatment has the added benefit of improved assay precision. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2373-8731 00000000 |
| Relation: | http://journals.lww.com/transplantationdirect/fulltext/10.1097/TXD.0000000000000709; https://doaj.org/toc/2373-8731 |
| DOI: | 10.1097/TXD.0000000000000709 |
| URL الوصول: | https://doaj.org/article/85b2e70e0d124f3a9a23fd0d816e846d |
| رقم الأكسشن: | edsdoj.85b2e70e0d124f3a9a23fd0d816e846d |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 23738731 00000000 |
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| DOI: | 10.1097/TXD.0000000000000709 |