Rahmah N, Anuar A K, A'shikin A N, Lim B H, Mehdi R, Abdullah B, Zurainee M N
School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
Biochem Biophys Res Commun. 1998 Sep 29;250(3):586-8. doi: 10.1006/bbrc.1998.9360.
Western blot analyses were performed on 444 serum specimens: 40 sera from microfilaraemic individuals, 10 sera from elephantiasis patients, 24 treated individuals, 50 sera from residents of endemic areas without anti-filarial IgG4 antibodies (endemic normals), 20 sera from amicrofilaraemic individuals with high anti-filarial IgG4 antibodies, 200 sera from healthy city-dwellers (non-endemic samples), and 100 sera from soil-transmitted helminth-infected individuals. Phast electrophoresis system was used to electrophorese Brugia malayi soluble adult worm antigen on 10-15% SDS-PAGE gradient gels followed by electrophoretic transfer onto PVDF membranes. Membrane strips were then successively incubated with blocking solution, human sera, and monoclonal anti-human IgG4 antibody-HRP, with adequate washings done in between each incubation step. Luminol chemiluminescence detection was then used to develop the blots. An antigenic band with the MW of approximately 37 kDa was found to be consistently present in the Western blots of all microfilaraemic sera, all amicrofilaraemic sera with high titres of anti-filarial IgG4 antibodies, some treated patients, and some elephantiasis patients. The antigen did not occur in immunoblots of individuals with other helminthic infections, normal endemic individuals, and city dwellers. Therefore the B. malayi antigen of with the MW of approximately 37 kDa demonstrated specific reactions with sera of B. malayi-infected individuals and thus may be useful for diagnostic application.
对444份血清标本进行了蛋白质免疫印迹分析：40份来自微丝蚴血症患者的血清、10份来自象皮肿患者的血清、24份已治疗个体的血清、50份来自流行区无抗丝虫IgG4抗体居民（流行区正常人）的血清、20份来自抗丝虫IgG4抗体水平高的无微丝蚴血症个体的血清、200份来自健康城市居民（非流行区样本）的血清以及100份来自土壤传播蠕虫感染个体的血清。使用Phast电泳系统在10 - 15% SDS - PAGE梯度凝胶上对马来布鲁线虫可溶性成虫抗原进行电泳，随后将其电转移至PVDF膜上。然后将膜条依次与封闭液、人血清以及单克隆抗人IgG4抗体 - HRP孵育，在每个孵育步骤之间进行充分洗涤。接着使用鲁米诺化学发光检测法显影印迹。在所有微丝蚴血症血清、所有抗丝虫IgG4抗体滴度高的无微丝蚴血症血清、部分已治疗患者以及部分象皮肿患者的蛋白质免疫印迹中，均持续出现一条分子量约为37 kDa的抗原条带。该抗原在其他蠕虫感染个体、流行区正常个体以及城市居民的免疫印迹中未出现。因此，分子量约为37 kDa的马来布鲁线虫抗原与马来布鲁线虫感染个体的血清呈现特异性反应，因而可能有助于诊断应用。
