Bessho T, Taira S, Ikuma K, Shigeta M, Koyama K, Isojima S
Nihon Sanka Fujinka Gakkai Zasshi. 1984 Mar;36(3):391-6.
In order to apply the early pregnancy factor (EPF) to early diagnosis of fertilization, the establishment of optimal conditions for assay of EPF was attempted, and then EPF in the sera of contracepted and conceived women 4 to 6 days after ovulation were measured. For assay of EPF, 0.25 ml of 1:2 step diluted anti-human lymphocyte serum (ALS), 0.05ml of guinea pig serum as complement and 0.1ml of lymphocytes suspension (1 X 10(7)/ml) pretreated with test serum were mixed and then incubated at 37 degrees C for 90 min. To this mixture 0.1ml of sheep red blood cell suspension (2 X 10(9)/ml) was added and the rosette formation was counted after centrifugation. The rosette inhibition titer (RIT) was expressed as reciprocal of ALS dilutions which resulted in less than 75% of rosette formation as compared with the control. RITs of the conceived women who were assayed on the 5 th day after ovulation were in the range from 16 to 32 X 10(3), while that of the control contraceptive women who were assayed on the same day was in the range from 2 to 4 X 10(3). The sterile women who received AIH but failed to conceive all showed less than 4 X 10(3) as RIT. These results suggest that the assay of EPF is valuable in detecting the early stage of fertilization and possibly may help to differentiate the impairment of embryo implantation from non-fertilization of the ovum as a cause of sterility.
为了将早孕因子（EPF）应用于受精的早期诊断，尝试建立EPF检测的最佳条件，然后测定排卵后4至6天避孕和受孕女性血清中的EPF。检测EPF时，将0.25ml 1:2逐步稀释的抗人淋巴细胞血清（ALS）、0.05ml豚鼠血清作为补体以及0.1ml经检测血清预处理的淋巴细胞悬液（1×10⁷/ml）混合，然后在37℃孵育90分钟。向该混合物中加入0.1ml绵羊红细胞悬液（2×10⁹/ml），离心后计数玫瑰花结形成情况。玫瑰花结抑制效价（RIT）表示为与对照组相比导致玫瑰花结形成少于75%的ALS稀释倍数的倒数。在排卵后第5天检测的受孕女性的RIT范围为16至32×10³，而在同一天检测的对照避孕女性的RIT范围为2至4×10³。接受人工授精但未受孕的不育女性的RIT均低于4×10³。这些结果表明，EPF检测在检测受精早期阶段有价值，并且可能有助于区分作为不育原因的胚胎着床障碍与卵子未受精。
