Oke Stacey L, Hurtig Mark B, Keates Robert A, Wright Jen R
Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada N1G 2W1.
Am J Vet Res. 2003 Jul;64(7):894-9. doi: 10.2460/ajvr.2003.64.894.
To develop a new 1,9-dimethylmethylene blue (DMMB) assay for measurement of sulfated glycosaminoglycan (sGAG) concentrations in equine synovial fluid (SF) by use of membrane technology and to compare the assay's ability to measure sGAG concentrations with that of 2 other established DMMB assays.
25 samples of SF collected from affected joints of 14 horses and 13 samples of SF collected from nonaffected (control) joints of 4 horses.
A solid-phase DMMB assay was developed to measure sGAG concentrations in SE Results for the assay were then compared with results obtained by use of the direct spectrophotometric method (ie, Famdale method) and microplate DMMB assay.
The solid-phase assay and direct spectrophotometric assay measured the same sGAG concentrations in identical equine SF, but those concentrations differed significantly from results obtained by use of the microplate DMMB assay. All other aspects of the solid-phase DMMB assay were comparable to both the direct spectrophotometric and microplate DMMB assays.
The new solid-phase assay can be used interchangeably with the direct spectrophotometric method to measure sGAG concentrations in equine SF samples, but it cannot be interchanged with the microplate DMMB assay. Results can be rapidly obtained with the solid-phase assay. Also, the solid-phase assay can detect nanogram quantities of sGAGs in SF, circumvent the problem of premature precipitation of sGAG-dye complexes, and provide quantitative or qualitative results. The solid-phase assay may replace other DMMB assays for measuring sGAG concentrations in SF obtained from horses.
开发一种新的1,9 - 二甲基亚甲基蓝（DMMB）检测方法，利用膜技术测定马滑膜液（SF）中硫酸化糖胺聚糖（sGAG）的浓度，并将该检测方法测定sGAG浓度的能力与另外两种已建立的DMMB检测方法进行比较。
从14匹马的患病关节采集25份SF样本，从4匹马的未患病（对照）关节采集13份SF样本。
开发一种固相DMMB检测方法来测定SF中的sGAG浓度。然后将该检测方法的结果与使用直接分光光度法（即Famdale法）和微孔板DMMB检测方法获得的结果进行比较。
固相检测方法和直接分光光度检测方法在相同的马SF中测定的sGAG浓度相同，但这些浓度与使用微孔板DMMB检测方法获得的结果有显著差异。固相DMMB检测方法的所有其他方面与直接分光光度法和微孔板DMMB检测方法相当。
新的固相检测方法可与直接分光光度法互换使用，以测定马SF样本中的sGAG浓度，但不能与微孔板DMMB检测方法互换。固相检测方法可快速获得结果。此外，固相检测方法可检测SF中纳克量的sGAG，避免sGAG - 染料复合物过早沉淀的问题，并提供定量或定性结果。固相检测方法可能会取代其他DMMB检测方法，用于测定从马身上获得的SF中的sGAG浓度。
