APPswe/PS1dE9转基因小鼠的剖腹产净化技术研究

目的 研究感染肠道鞭毛虫转基因小鼠的生物净化技术.剖腹产方法对感染的小鼠进行净化,使其达到SPF级标准,为建立基因工程小鼠肠道鞭毛虫净化技术提供依据.方法 取感染肠道鞭毛虫的APPswe/PS1dE9阿尔茨海默病（Alzheimer＇s disease,AD）转基因小鼠（APP小鼠）,在超净工作台内行剖腹产取胎手术,用SPF级帕金森病转基因鼠代乳,术后1、4、7、10月进行微生物和寄生虫检测;PCR技术鉴定剖腹所得APP仔鼠及其繁殖群（F1代）APPswe基因型,计算F1代每窝仔鼠APPswe基因阳性率及平均阳性率,用单样本t检验分析APPswe基因阳性率是否符合孟德尔遗传规律.结果 实施剖腹产手术10例,获仔鼠75只,7日龄仔鼠存活率及离乳存活率分别为90.67%（68/75）和73.33%（55/75）;净化前APP小鼠肠道鞭毛虫感染率均为100%（5/5）,净化后的4次检测,APP小鼠SPF级所要求的病原菌及寄生虫结果均为阴性.净化后的APP仔鼠共繁殖42窝F1代,平均每窝APPswe基因阳性率为48.3%,与孟德尔遗传规律50%的理论阳性值相比无统计学差异（P＞0.05）.结论 剖腹产净化技术可成功去除该APP小鼠的肠道鞭毛虫,且对APPswe基因遗传性状未产生明显影响.

Study of Decontamination Technique among APPswe/PS1dE9 Transgenic Mice using Caesarian Section

Objective To study decontamination technique of caesarean section among flagellates infected transgenic mice, and to establish specific pathogen-free （SPF） mice strain in order to provide technical support for establishment of flagellates-eradication methods of genetically engineered mice. Methods APPswe/PSldE9 transgenie mice （APP mice） were used to conduct this research. Caesarean section was taken on pregnant APP mice to get pups under a SPF barrier environment, and the pups were transferred to female SPF Parkinson＇s Disease transgenic mice which served as a foster mother. Survival rates of pups were calculated in 7 days of age and after weaning. Pathogens were tested in the first month, and later, every 3 months since Caesarean sections. APPswe gene of APP mice was amplified by polymerase chain reaction （PCR） for genotyping. Average gene positive rate was calculated among the pups and their offspring （F1） , and one sample t test was conducted to compare the average gene positive rate with the theoretical Mendelian segregationratio （50%）. Results We performed 11 Caesarean sections on APP pregnant mice, and a total of 75 pups were collected and transferred to a foster mother. Survival rate of the pups after 7 days and weaning was 90. 67% （68/75） and 73.33% （55/75） , respectively. Pathogens tested before decontamination showed flagellates infection among APP mice was 100% （5/5） , but after decontamination, all the pathogens required for SPF all show negative results. The average APPswe gene positive rate of the dams in APP mice after decontamination was 48.3% , which was in accordance with the Mendelian segregation ratio （ P 〉 0.05 ）. Conclusion We had successfully eradicated flagellates from the APP mice by caesarean section and a foster mother technique with stable hereditary feature of APPswe gene.