Sharmin S, Ahmed S, Abu Saleh A, Rahman F, Choudhury M R, Hassan M M
Bangladesh Med Res Counc Bull. 2014 Aug;40(2):74-8. doi: 10.3329/bmrcb.v40i2.25225.
Antinuclear antibody (ANA) is useful in the diagnosis of connective tissue disorder (CTD). Association of specific autoantibodies with the immunofluorescence pattern of ANA in CTD, noted in western literature has been considered as reference in all over the world. However, in Bangladesh no such research work or data correlating the autoantibodies and their ANA patterns is found. Objective of the study was to identify an association between immunofluorescence patterns of antinuclear antibody on HEp-2 cell and more specific antinuclear reactivities (e.g. anti-dsDNA and anti-extractable nuclear antigen) in the serum samples of CTD patients. Serum samples of 152 CTD patients (Systemic lupus erythematosus, Rhumatoid arthritis, Sjogren's syndrome, Systemic sclerosis, Polymyositis, Mixed connective tissue disease) were diagnosed clinically, attending at Bangabandhu Sheikh Mujib Medical University (BSMMU) during the study period of January, 2010 to December, 2010. Samples were subjected for ANA testing by Indirect Immunofluorescence (IIF) on HEp-2 cell (ALPHADIA) in dilution of 1:40, anti-dsDNA by ELISA and anti- extractable nuclear antigen (anti-ENA) by Dot Immunoblot. Dot blot strips were tested for anti-Sm, anti-RNP, anti-SSA/Ro, anti-SSB/La, anti-Scl-70 and anti-Jo-1. Out of 152 patients 110 (72.3%) cases were ANA positive by IIF on HEp-2 cell. ANA positive sera exhibited four fluorescence patterns such as speckled (50.8%), peripheral (21.6%) , homogenous (18.1%) and nucleolar pattern (9%). Peripheral pattern and homogenous pattern was predominantly associated with anti-dsDNA (p < 0.05). Speckled pattern was significantly associated with anti-ENA (p < 0.05).The most commonly identified antinuclear autoreactivity was directed towards anti-RNP (25.7%) then anti-Scl-70 (20%), anti-SSA (14.2%) and anti-SSB (5.7%). Multiple anti-ENA reactivities were identified in 34.28% cases. Peripheral and homogenous pattern is strongly associated with anti-dsDNA and speckled pattern may predict anti-ENA (specially ribonucleoprotiens). As a definite correlation between the ANA patterns and the group of antibodies was detected by dot immunoblot, one could predict presence of certain specific auto antibodies for a particular ANA pattern identified. This may restrict on the cost of laboratory investigations in a developing country like Bangladesh. Thus, ANA-IIF method may reduce the expense of detailed immunological work-up with minimal loss in diagnostic accuracy.
抗核抗体(ANA)在结缔组织病(CTD)的诊断中很有用。西方文献中提到的特定自身抗体与CTD中ANA免疫荧光模式的关联在全世界都被视为参考。然而,在孟加拉国,尚未发现将自身抗体及其ANA模式相关联的此类研究工作或数据。本研究的目的是确定CTD患者血清样本中HEp-2细胞上抗核抗体的免疫荧光模式与更特异性的抗核反应性(如抗双链DNA和抗可提取核抗原)之间的关联。在2010年1月至2010年12月的研究期间,对在班加班杜·谢赫·穆吉布医科大学(BSMMU)就诊的152例CTD患者(系统性红斑狼疮、类风湿性关节炎、干燥综合征、系统性硬化症、多发性肌炎、混合性结缔组织病)的血清样本进行了临床诊断。样本通过间接免疫荧光法(IIF)在稀释度为1:40的HEp-2细胞(ALPHADIA)上进行ANA检测,通过酶联免疫吸附测定法检测抗双链DNA,通过斑点免疫印迹法检测抗可提取核抗原(抗ENA)。对斑点印迹条带检测抗Sm、抗RNP、抗SSA/Ro、抗SSB/La、抗Scl-70和抗Jo-1。在152例患者中,110例(72.3%)通过HEp-2细胞上的IIF检测ANA呈阳性。ANA阳性血清表现出四种荧光模式,如斑点状(50.8%)、周边型(21.6%)、均质型(18.1%)和核仁型(9%)。周边型和均质型主要与抗双链DNA相关(p<0.05)。斑点状模式与抗ENA显著相关(p<0.05)。最常见的抗核自身反应性是针对抗RNP(25.7%),其次是抗Scl-70(20%)、抗SSA(14.2%)和抗SSB(5.7%)。在34.28%的病例中发现了多种抗ENA反应性。周边型和均质型与抗双链DNA密切相关,斑点状模式可能预示抗ENA(特别是核糖核蛋白)。由于通过斑点免疫印迹法检测到ANA模式与抗体组之间存在明确的相关性,因此可以预测特定ANA模式下某些特定自身抗体的存在。这可能会限制像孟加拉国这样的发展中国家实验室检查的成本。因此,ANA-IIF方法可能会在诊断准确性损失最小的情况下降低详细免疫检查的费用。