Makondo K, Amiridis G S, Jeffcoate I A, O'Shaughnessy P J, Boyd J S, Paterson C, Robertson L
Department of Veterinary Anatomy and Physiology, Glasgow Veterinary School, UK.
Anim Reprod Sci. 1997 Dec 5;49(2-3):125-33. doi: 10.1016/s0378-4320(97)00071-7.
The aim of this study was to carry out first trimester fetal sex diagnosis using the polymerase chain reaction (PCR) to amplify DNA from bovine fetal cells recovered by transvaginal ultrasound-guided uterine puncture and fetal fluid aspiration. For sex determination, a nested, allele-specific, PCR amplification of the bovine zfx and zfy gene fragments was utilised. The PCR assay was validated using fetal fluids recovered from uteri post mortem. Cells were harvested from the fetal fluids, genomic DNA extracted and the PCR assay applied. A technique which was developed for transvaginal ultrasound-guided follicle aspiration was modified to recover fetal fluid from live animals. Small volumes of fetal fluid (0.5-5 ml) were recovered between days 61-97 of gestation and the PCR assay applied. The gender determined by PCR of fetal fluid cells was in all cases confirmed by visual inspection (n = 15 abattoir specimens) or ultrasound scanning (n = 7 live animals). Fetal death, attributed to the introduction of intrauterine infection, occurred in 4/4 cows in the first series of aspirations but in only 1/3 heifers in the second series of aspirations.
本研究的目的是利用聚合酶链反应（PCR）对经阴道超声引导下子宫穿刺和羊水抽吸所获得的牛胎儿细胞中的DNA进行扩增，以实现孕早期胎儿性别诊断。为了确定性别，采用了对牛zfx和zfy基因片段进行巢式、等位基因特异性PCR扩增的方法。使用从死后子宫中回收的羊水对PCR检测进行验证。从羊水中采集细胞，提取基因组DNA并进行PCR检测。对一种为经阴道超声引导下卵泡抽吸而开发的技术进行了改进，以从活体动物中回收羊水。在妊娠第61 - 97天之间回收少量羊水（0.5 - 5毫升）并进行PCR检测。通过对羊水细胞进行PCR确定的性别在所有情况下均通过肉眼检查（n = 15份屠宰场标本）或超声扫描（n = 7头活体动物）得到证实。在第一系列抽吸中，4/4头母牛因引入宫内感染而发生胎儿死亡，但在第二系列抽吸中，仅1/3头小母牛出现这种情况。
