非同位素标记pAW101探针作人流胚胎组织的亲权鉴定

PATERNITY DETERMINATION OF ABORTED FETUS USING NONRADIO LABELED PAW101 PROBE

DNA限制性片段长度多态性(RFLPs)技术的经典方法是用酚抽提DNA,同位素标记探针。本文研究选用N_aCI盐析分离DNA的技术,结合地高辛配基标记pAW101探针方法,作了11例无争议的胚胎组织的亲权鉴定,同时测定4种同Ⅰ酶型,在不违反孟德尔遗规律的前提下,按Essen-Moller公式计算父权概率(均>99.73％),达到确定亲生关系的标准。本方法简单、易于掌握,结果稳定可靠,适宜于一般实验室开展工作。

The classical technique determining restriction fragment length polymorphisms(RFLPs) of human DNA involves the procedures of extracting DNA with phynol and labeling probe with radio isotope. It requires well-equipped-conditions, rather complicate processes, and may be affected by the supply and the half life of radio isotope. Besides, the phynol, chloroform, and radio isotope are harmful to human being and polluting environment. 1 his paper reports the combination isolating DNA using NaCl salting out method with labeling pAW 101 probe using Digoxigenin-dUTP to show RFLPs. With this method, the paternity testing of aborted feta were performed. Meanwhile, the phenotypes of four isosnzymes, phosphoglucomutase, esterase D, glyoxylase I, and acid phosphatase, were tested. According to the occurence frequency of RF and of the enzymes in the population of Guangdong, China, the probability of paternity (W value) were caculated by the standard procedure described by Essen-Moller. The results showed that the W value of each case was over 99.73 %, reaching the standard of including paternity. It is a rapid, effective, economic and non -harmful RFLPs technique, suits common laboratory performance and is worth whidely to spread.