有限稀释微量溶血法测定小鼠血清溶血素的活性  

Determination of mouse serum hemolysin by limited dilution combined with micro hemolysis

作　　者：雷林生[1] 孙莉莎[1] 杨淑琴[1] 

机构地区：[1]第一军医大学药理学教研室

出　　处：《中国药理学通报》1999年第3期274-276,共3页Chinese Pharmacological Bulletin

摘　　要：目的检测抗绵羊红细胞（ＳＲＢＣ）抗体（溶血素）。方法将含有溶血素的血清样品适当预稀释，然后取１００μｌ加入９６孔板中进行有限的倍比稀释，每孔再加入体积分数为１５％ＳＲＢＣ和１∶１０稀释的豚鼠血清各５０μｌ，在３７℃温育１～１２ｈ后，有足够溶血素活性孔的红细胞将全部溶解，小孔液体清彻见底；溶血素活性低于足量时，只能部分溶血，受未溶解红细胞的阻挡，小孔底部只有不同程度的透明度，将印有字母“Ｒ”的测试纸放在９６孔板底部，根据字母“Ｒ”的能见程度和已知的稀释倍数判断样品的活性。结果样品的测定值与理论值成良好直线关系。对已知药物的效果测定与用分光光度法所测得结果一致。结论本方法简单、快速、准确、重复性好。AIM To determine the activities of antibodies (hemolysin) against sheep red blood cells (SRBC). METHOD Serum samples containing hemolysin were pre diluted. Then 100 μl of each diluted samples was transferred into the wells of a 96 well plate for further limited serial dilution with two fold each time. After that each well received 50 μl of 1 5% SRBCs and 1∶10 diluted guinea pig sera respectively. After incubation for 1 to 12h at 37℃ the SRBCs in the wells containing enough amount of hemolysin were completely lysed and the bottoms of the wells could be thoroughly transparent. However, the SRBCs in the wells containing less amounts of hemolysin were only partially lysed and with the blockade of the remaining intact SRBCs the transparency of the bottoms was impared to some extent. The hemolysin activities of samples were determined by the combination of the transparent degrees of the wells with the known diluted times. Under the plate a test sheet on which the capital letter “R”s had been printed was placed for transparency determination. RESULTS The assessed values were well lineally correlated with the theoretical ones and the results obtained by the method for several drugs were in accordance with those by spectrophotometry.CONCLUSION The procedure of the method is simple and the results are quickly obtained, accurate and reproducible.

关 键 词：抗体 溶血素 微量测定 免疫调节药物 药物筛选 

分 类 号：R979.5[医药卫生—药品] R965.1[医药卫生—药学]