Krajčíková Kristína, Glinská Gabriela, Tomečková Vladimíra
Department of Medical and Clinical Biochemistry, Faculty of Medicine, Pavol Jozef Šafárik University in Košice, Košice, Slovakia.
The Ophthalmologic Practice PROOKO, Košice, Slovakia.
Taiwan J Ophthalmol. 2021 Jun 15;12(1):88-92. doi: 10.4103/tjo.tjo_14_21. eCollection 2022 Jan-Mar.
Human tears contain more than 1500 proteins that could be diagnostically relevant. To date, numerous candidates on a biomarker of protein origin were identified for ocular and systemic diseases. However, the suitable sampling method is still the subject of discussion. To address the need for a description of sampling methods properties for possible clinical analyses, we studied a total protein concentration and electrophoretic pattern of tear fluid collected by capillary tubes, Schirmer strips, cellulose microsponges, and flushing. The total protein concentration was 4.339 μg/μL ± 1.905 μg/μL, 0.967 μg/μL ± 0.117 μg/μL, 0.022 μg/μL ± 0.016 μg/μL, and 0.008 μg/μL ± 0.006 μg/μ for the capillary tubes, Schirmer strips, flushing, and cellulose microsponges, respectively. Sodium dodecyl sulfate polyacrylamide electrophoresis showed the different patterns of tear proteins obtained by the above-mentioned sampling methods. These differences could originate from the use of a bigger amount of extraction reagent that was not used in the case of capillary tubes, and retention of the proteins by strips and sponges. Taken together, capillary tubes, Schirmer strips, cellulose microsponges, and flushing represent sensitive and convenient sampling methods for tear fluid collection. For the isolation of proteins from strips and sponges, and for the flushing, less than 100 μL of a reagent should be used to ensure the sufficient concentration of the biomarkers in a trace amount.
人类眼泪含有1500多种可能具有诊断意义的蛋白质。迄今为止，已鉴定出许多蛋白质来源的眼部和全身性疾病生物标志物候选物。然而，合适的采样方法仍是讨论的主题。为满足对可能的临床分析中采样方法特性描述的需求，我们研究了用毛细管、泪液试纸、纤维素微海绵和冲洗法收集的泪液的总蛋白浓度和电泳图谱。毛细管、泪液试纸、冲洗法和纤维素微海绵收集的泪液总蛋白浓度分别为4.339μg/μL±1.905μg/μL、0.967μg/μL±0.117μg/μL、0.022μg/μL±0.016μg/μL和0.008μg/μL±0.006μg/μL。十二烷基硫酸钠聚丙烯酰胺电泳显示了上述采样方法获得的泪液蛋白质的不同图谱。这些差异可能源于使用了比毛细管法更多的提取试剂，以及试纸和海绵对蛋白质的保留。综上所述，毛细管、泪液试纸、纤维素微海绵和冲洗法是收集泪液的灵敏且方便的采样方法。从试纸和海绵中分离蛋白质以及冲洗时，应使用少于100μL的试剂，以确保痕量生物标志物有足够的浓度。
