Noskova Libuse, Fialova Lenka, Bartos Ales, Zima Tomas
Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine, Charles University and General University Hospital in Prague, Czech Republic.
National Institute of Mental Health, Klecany, Czech Republic.
Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2017 Jun;161(2):179-186. doi: 10.5507/bp.2017.017. Epub 2017 Apr 26.
To optimise the ELISA method for the avidity of IgG antibodies against neurofilament heavy chain (NfH) and to determine the levels and avidity of anti-NfH antibodies in patients with Alzheimer's disease (AD) and a healthy control group.
Various dilutions of sera and concentrations of urea and sodium chloride as chaotropic reagents were tested in the process of the ELISA optimisation. The levels and avidity of anti-NfH antibodies were determined in 30 patients with Alzheimer's disease and 30 age-matched cognitively normal elderly adults.
Sera dilution 1:200 and urea as a chaotrope in a concentration 6 mol/L were chosen to be the most suitable for the avidity assay of anti-NfH antibodies by ELISA. The results showed no differences in either level or avidity of IgG anti-NfH antibodies between AD patients and cognitively normal persons. The levels of anti-NfH IgG antibodies inversely correlated with their avidities.
We optimised the ELISA method for the determination of anti-NfH antibody avidity determination which is suitable for research of anti-NfH antibody avidity in patients with neurological diseases associated with neurocytoskeletal defects. The determination of serum anti-NfH antibody avidity in AD patients seems to have limited diagnostic significance.
优化用于检测抗神经丝重链(NfH)IgG抗体亲和力的ELISA方法,并测定阿尔茨海默病(AD)患者和健康对照组中抗NfH抗体的水平及亲和力。
在ELISA优化过程中,测试了血清的各种稀释度以及作为离液剂的尿素和氯化钠的浓度。测定了30例阿尔茨海默病患者和30例年龄匹配的认知正常老年人中抗NfH抗体的水平及亲和力。
选择血清稀释度1:200和浓度为6 mol/L的尿素作为离液剂最适合通过ELISA检测抗NfH抗体的亲和力。结果显示,AD患者和认知正常者之间IgG抗NfH抗体的水平和亲和力均无差异。抗NfH IgG抗体的水平与其亲和力呈负相关。
我们优化了用于测定抗NfH抗体亲和力的ELISA方法,该方法适用于研究与神经细胞骨架缺陷相关的神经系统疾病患者的抗NfH抗体亲和力。测定AD患者血清抗NfH抗体亲和力的诊断意义似乎有限。