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当前位置: 首页 > 文章 > 应用母体外周血中游离DNA鉴定细毛羊胎儿性别研究 中国畜牧杂志 2016,52 (23) 10-14
Position: Home > Articles > Detection of Fine-wool Sheep Fetal Sex From Cell-free Fetal DNA in Maternal Peripheral Blood Chinese Journal of Animal Science 2016,52 (23) 10-14

应用母体外周血中游离DNA鉴定细毛羊胎儿性别研究

作  者：
赵冰茹;黄锡霞;田可川;石秋香;田月珍;石晓雷
单  位：
新疆农业大学动物科学学院;新疆畜牧科学院;新疆绒毛用羊遗传育种与繁殖实验室
关键词：
细毛羊;外周血游离DNA;SRY;性别鉴定
摘  要：
本研究针对不同妊娠时期细毛羊进行胎儿性别鉴定工作。首先对外周血浆/血清分别采用加热法和酚-氯仿法提取游离DNA。通过普通PCR扩增雄性性别决定基因(sex determining region Y,SRY)和β-actin检测基因的目的片段。鉴于部分样品未扩增出SRY基因目的片段,设计2对内外引物,运用优化后的巢式PCR体系再次针对SRY目的基因展开特异性扩增。最后对每个妊娠时间段的鉴定结果和实际生产结果进行统计,计算准确率。结果表明:加热法和酚-氯仿法均成功分离提取出妊娠母羊外周血浆/血清中的游离DNA,并扩增出β-actin检测基因片段;SRY阳性样本扩增出了SRY目的基因片段和β-actin检测基因片段,SRY阴性样本只扩增出了β-actin检测基因片段;性别鉴定的平均准确率达到81.25%(19.5/24),SRY基因检出率与妊娠时间的长短呈正比。结论,本研究建立了一种基于外周血游离DNA检测细毛羊胎儿性别的非创伤性且高效低廉的产前诊断技术,但性别鉴定的准确率还有待提高。
译  名：
Detection of Fine-wool Sheep Fetal Sex From Cell-free Fetal DNA in Maternal Peripheral Blood
作  者：
ZHAO Bing-ru;HUANG Xi-xia;TIAN Ke-chuan;SHI Qiu-xiang;TIAN Yue-zhen;SHI Xiao-lei;College of Animal Science, Xinjiang Agricultural University;The Laboratory of Genetics, Breeding and Reproduction for Xinjiang Wool & Cashmere Sheep;Institute of Animal Science, Xinjiang Academy of Animal Sciences;
关键词：
fine-wool Sheep;;peripheral blood free DNA;;SRY;;sex identification
摘  要：
The research aimed at fine-wool Sheep of different gestation period to implement f etal sex identification work. Firstly, using the heating method and phenol chloroform method extract free DNA in peripheral blood plasma/serum respectively. Amplified fragment length of male sex determination gene(sex determining region Y, SRY) and β-actin detecting gene for ordinary PCR technology was wsed. In view of some samples not amplified fragments of SRY gene, two inner and outer primers were designed and optimal system of nested PCR on SRY gene for specific amplification fragments was wsedonce again. Finally, thedata about identification results of each gestation period and actual production results were collectedto calculate accuracy. The results showed that heating method and phenol chloroform method were successfully extractedin peripheral blood plasma/serum of free DNA onpregnancy sheep and amplified fragment of β-actin detecting gene, SRY positive samples amplified fragments of the SRY gene and β-actin gene, while SRY negative samples was amplified fragments of only β-actin gene. The average accuracy of sex determination was 81.25%(19.5/24), SRY detection rate was positively with pregnancy days. In conclusion, the study establishesd a non-traumatic, efficient and cheap prenatal diagnosis technology based on peripheral blood of free DNA, but the accuracy of sex determination needs to be improved.

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