Yilmaz Ozlem, Karaman Meral, Ergon M Cem, Bahar I Hakkı, Yuluğ Nuran
Dokuz Eylül Üniversitesi Tıp Fakültesi Mikrobiyoloji ve Klinik Mikrobiyoloji Anabilim Dalı, İnciraltı, İzmir, Turkey.
Turkiye Parazitol Derg. 2005;29(4):287-90.
Connective tissue diseases (CTD) are characterized by the presence of autoantibodies against several tissues. These autoantibodies occur against cell membrane, cell receptors, plasma proteins, and cytoplasmic and nuclear components. In laboratories, anti-nuclear antibodies (ANA) and anti-double stranded DNA (anti-dsDNA) antibodies are widely used in diagnosis of CTD. The aim of this study was to investigate the presence and accompaniment of ANA and anti-dsDNA antibodies in diagnosis of several CTD and also to study the prevalence of ANA and anti-dsDNA in a group of 88 patients with various types of CTD. ANA were detected by immunofluorescence (IFA) using HEp-2 cells (Zeus Scientific, Inc. USA) and anti-dsDNA antibodies using Crithidia luciliae (BioSystems, Spain) as substrates in immunofluorescence. ANA Western Blot (WB) Immunoassay (ImmuBlot, International Immuno-Diagnostics, USA) was also used along with the tests referred to previously. ANA was found in the sera of 84 (96.5%) patients while anti-dsDNA was detected in 7 (7.95%). Moreover different fluorescence patterns were also evaluated with ANA IFA in accordance with anti-dsDNA results. Mixed patterns in three and a homogeneous pattern in four anti-dsDNA positive patients' sera were determined on HEp-2 cell line by IFA. Seven sera which were ANA and anti-dsDNA positive with IFA were also found to be positive with WB and their ANA patterns with the specific ANA WB bands were also evaluated. It was observed that IFA results were in concordance with WB results. Our data indicated that the above findings should be controlled and evaluated with a more advanced method such as western blotting technique in order to confirm the presence of specific antibodies along with clinical outcome of the patients. As a result we think that ANA WB method is an appropriate technique in diagnosis of CTD as anti-dsDNA and ANA bands can be evaluated together with this method.
结缔组织病(CTD)的特征是存在针对多种组织的自身抗体。这些自身抗体针对细胞膜、细胞受体、血浆蛋白以及细胞质和细胞核成分产生。在实验室中,抗核抗体(ANA)和抗双链DNA(抗dsDNA)抗体被广泛用于CTD的诊断。本研究的目的是调查ANA和抗dsDNA抗体在几种CTD诊断中的存在情况及伴随情况,并研究88例不同类型CTD患者中ANA和抗dsDNA的患病率。采用免疫荧光法(IFA),以人喉癌上皮细胞(HEp-2细胞,美国宙斯科学公司)检测ANA,以利什曼原虫(西班牙生物系统公司)为底物,采用免疫荧光法检测抗dsDNA抗体。还使用了ANA蛋白质印迹(WB)免疫测定法(免疫印迹法,美国国际免疫诊断公司)以及之前提到的检测方法。84例(96.5%)患者血清中检测到ANA,7例(7.95%)检测到抗dsDNA。此外,还根据抗dsDNA结果,用ANA免疫荧光法评估了不同的荧光模式。通过免疫荧光法在HEp-2细胞系上确定了3例抗dsDNA阳性患者血清中的混合模式和4例中的均质模式。7例免疫荧光法检测ANA和抗dsDNA均为阳性的血清,蛋白质印迹法检测也为阳性,同时还评估了其具有特异性ANA蛋白质印迹条带的ANA模式。观察到免疫荧光法结果与蛋白质印迹法结果一致。我们的数据表明,上述发现应以更先进的方法如蛋白质印迹技术进行对照和评估,以便在确认患者临床结果的同时确定特异性抗体的存在。结果我们认为,ANA蛋白质印迹法是CTD诊断中的一种合适技术,因为该方法可同时评估抗dsDNA和ANA条带。