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[使用Synpor膜过滤器通过吲哚DNA过滤法分离和测定奶牛乳汁中的体细胞数量]

[Use of the Synpor membrane filter for the separation and determination of the number of somatic cells in the milk of dairy cows using the indole DNA filtration method].

作者信息

Urbanová E, Sedinová V, Skarda J, Skardová O, Jukl A

出版信息

Vet Med (Praha). 1985 Jul;30(7):409-18.

PMID:3927565
Abstract

The somatic cells of cow's milk were separated by filtering through nitrocellulose membrane filters (Synpor), pore size 2 to 5 micron. An indole reagent was added to the membrane filters with trapped cells, the mixture was warmed in boiling water bath for 20 min., then cooled, centrifuged (3500 X g, 15 min.), and optical density was measured at 490 nm. The relation 8 micrograms = 1 million cells was used for the conversion of DNA content in sample to the counts of cells. The average variation coefficient of the determination was 4.7% and the coefficient of correlation between the indole DNA filter method and direct microscopy on membrane filters was r = 0.997. Using the indole DNA filter method, the counts of somatic cells can be estimated from milk samples kept frozen or from filters with retained cells kept at 4 degrees C, or (for a short time--3 days) at 25 degrees C. The estimation is not distorted when the milk samples are stabilized with K2Cr2O7 or a mixture of K2Cr2O7 and HgCl2, if they are kept at 4 degrees C for eight days.

摘要

通过孔径为2至5微米的硝酸纤维素膜过滤器(Synpor)过滤,分离牛奶中的体细胞。将吲哚试剂添加到截留了细胞的膜过滤器中,混合物在沸水浴中加热20分钟,然后冷却、离心(3500×g,15分钟),并在490nm处测量光密度。采用8微克 = 100万个细胞的关系,将样品中的DNA含量换算为细胞计数。测定的平均变异系数为4.7%,吲哚DNA过滤法与膜过滤器直接显微镜检查之间的相关系数r = 0.997。使用吲哚DNA过滤法,可以从冷冻保存的牛奶样品、保存在4℃的截留细胞的过滤器或(短时间——3天)保存在25℃的样品中估计体细胞计数。如果牛奶样品用K2Cr2O7或K2Cr2O7与HgCl2的混合物稳定化,并在4℃保存8天,则估计结果不会失真。

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