生乳中体细胞快速检测技术的对比研究

Comparative study on rapid detection of somatic cells in raw milk

目的建立一种检测生乳中体细胞含量的快速检测方法(试剂盒法)。方法本试剂盒法通过生物酶溶解奶样中的蛋白质,缓冲液稳定脂肪球和蛋白并改变体细胞通透性,细胞裂解液及荧光染料快速渗透进入体细胞并沾染DNA,培养过程中,染液进入DNA的双螺旋结构,与DNA结合后荧光量子产率明显提高,最后通过荧光信号探测系统捕捉荧光强度来测定生乳中的体细胞数量。结果该方法与快速仪器法对比,相关系数达94%以上,2种方法在统计学上无显著性差异(P>0.05)。与标准方法对比,针对同一样品的检测结果的Log差值小于0.25, 2种方法统计学上也无显著性差异(P>0.05)。对不同样品进行重复性检测,相对标准偏差均小于15%。结论该试剂盒法具有较高的准确度和精密度,检测结果与标准方法和市售仪器快速检测法无明显差异,检测时间缩短50%,提升了检测效率。由于无需大型的检测设备及配套试剂,适合牧场和企业实验室的快速定量检测。

Objective To establish a rapid detection method(kit method) for the content of somatic cells in raw milk. Methods In this kit method, proteins in milk samples were dissolved by biological enzymes. The buffer solution stabilized fat globules and proteins, and changed the permeability of somatic cells. Cell lysate and fluorescent dyes quickly penetrated into the somatic cells and stained with DNA. During the culture process, the dye solution entered the double helix structure of DNA. After binding with DNA, the fluorescence quantum yield was significantly increased. Finally, the fluorescence intensity was captured by fluorescence signal detection system to determine the number of somatic cells in raw milk. Results Compared with the rapid instrument method, the correlation coefficient was over 94%, and there was no significant difference between the two methods(P>0.05). The Log difference for the same sample was less than 0.25 compared with the standard method, and there was no significant difference between the two methods statistically(P>0.05). When the repeatability of different samples was tested, the relative standard deviation were less than 15%. Conclusion The rapid detection technology has high accuracy and precision, and there is no significant difference of the results between the standard method and the rapid detection method of the market instrument. The detection time is shortened by 50%, and the detection efficiency is improved. Because no large-scale testing equipment and matching reagents are needed, it is suitable for rapid quantitative detection in pastures and enterprise laboratories.