Engineer L, Johnson R E, Bhol K C, Ahmed A R
Department of Oral Medicine and Diagnostic Sciences, Harvard School of Dental Medicine, Boston, MA 02115, USA.
Exp Dermatol. 2000 Aug;9(4):271-4. doi: 10.1034/j.1600-0625.2000.009004271.x.
Immunoblot assays have been developed to characterize the autoantigens and to detect autoantibodies in muco-cutaneous autoimmune vesiculo-bullous diseases using different substrates. However the results have been inconsistent, because availability and standardization of different substrates has been a major problem. The aim of this study was to develop an immunoblot assay using bovine gingival lysate as substrate because it is easily and readily available as well as inexpensive. Sera from patients with different vesiculo-bullous diseases were studied. These included 25 patients with pemphigus vulgaris (PV), 8 with paraneoplastic pemphigus (PNP), 12 with pemphigus foliaceus (PF), 25 with bullous pemphigoid (BP), and 22 with cicatricial pemphigoid (CP). Serum samples from 40 normal human volunteers were also studied. The autoantibody titers were determined based on the binding pattern of each disease and compared to those obtained by routine indirect immunofluorescence (IIF). Our observations suggest that the titers from immunoblot assays were significantly higher than titers obtained by IIF (P<0.0001). When the autoantibody titers were compared using bovine gingival lysate and human epidermal lysate as substrate, statistically significant differences were not observed. The use of bovine gingival lysate as a substrate will facilitate the rapid and early serological diagnosis of patients with vesiculobullous diseases. It may also be of benefit to laboratory investigators studying these autoantibodies.
免疫印迹分析已被开发出来，用于鉴定自身抗原，并使用不同底物检测黏膜皮肤自身免疫性水疱大疱性疾病中的自身抗体。然而，结果并不一致，因为不同底物的可用性和标准化一直是一个主要问题。本研究的目的是开发一种以牛牙龈裂解物为底物的免疫印迹分析方法，因为它易于获得、价格低廉。研究了不同水疱大疱性疾病患者的血清。这些患者包括25例寻常型天疱疮（PV）、8例副肿瘤性天疱疮（PNP）、12例落叶型天疱疮（PF）、25例大疱性类天疱疮（BP）和22例瘢痕性类天疱疮（CP）。还研究了40名正常人类志愿者的血清样本。根据每种疾病的结合模式确定自身抗体滴度，并与通过常规间接免疫荧光（IIF）获得的滴度进行比较。我们的观察结果表明，免疫印迹分析的滴度显著高于IIF获得的滴度（P<0.0001）。当以牛牙龈裂解物和人表皮裂解物为底物比较自身抗体滴度时，未观察到统计学上的显著差异。使用牛牙龈裂解物作为底物将有助于水疱大疱性疾病患者的快速和早期血清学诊断。这对研究这些自身抗体的实验室研究人员也可能有益。
