软骨中DNA的荧光测定及基质成分干扰的校正
Fluorometric assay of DNA in cartilage and correction for interference by matrix components.
作者信息
Miller D R, Mankin H J
出版信息
Connect Tissue Res. 1981;9(2):89-94. doi: 10.3109/03008208109160245.
A fluorometric assay using 3,5-diaminobenzoic acid for analysis of DNA in alkaline-hydrolyzed calf articular cartilage is described. Additional estimates of DNA content, obtained by counting of cells in cartilage slices and of cells isolated from the tissue by digestion with bacterial collagenase, indicated that falsely high values were obtained with the fluorometric assay and with the indole procedure of Bonting and Jones. Cartilage collagen and proteoglycan were both found to contribute to this interference. Factors to correct for this interference were determined, based upon the hydroxyproline and uronic acid contents of these components. application of these corrections to DNA values obtained by fluorometric assay of the cartilage samples resulted in DNA estimates in agreement with those obtained by counting of cells.
描述了一种使用3,5 - 二氨基苯甲酸的荧光测定法，用于分析碱性水解的小牛关节软骨中的DNA。通过对软骨切片中的细胞以及用细菌胶原酶消化组织后分离出的细胞进行计数，获得了DNA含量的额外估计值，结果表明，荧光测定法和邦廷与琼斯的吲哚法都得出了错误的高值。发现软骨胶原蛋白和蛋白聚糖均对此干扰有影响。基于这些成分的羟脯氨酸和糖醛酸含量，确定了校正此干扰的因素。将这些校正应用于通过荧光测定法获得的软骨样品的DNA值，得到的DNA估计值与通过细胞计数获得的估计值一致。
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