基因治疗奶牛乳腺炎的生物安全评价

Biosafety evaluation of gene therapy on bovine subclinical mastitis

为评价基因治疗奶牛乳腺炎的生物安全性,以3头患隐性乳腺炎的奶牛作为试验对象,通过乳房基部穿刺注射接种携带人溶菌酶基因的乳腺特异性表达重组质粒p215C3LYZ(1 600μg.头-1),24 h后以相同剂量重复注射1次。定期采集试验奶牛的乳样、粪样、尿样以及试验牛舍的垫料和水样,并利用建立的PCR进行检测。结果发现,除第2次注射24 h内可从奶样中检测到重组质粒外,其他样品均为阴性,这表明重组质粒p215C3LYZ不能在奶牛体内长期残留,也不能在环境中释放及散播。所有奶牛均于第2次注射20 d后处死,并采集血液、心、肝、脾、肺、肾、肌肉以及乳腺等样品进行PCR检测,结果表明:重组质粒p215C3LYZ在奶牛体内未发生转移。

To evaluate the clinical biosafety of gene therapy on bovine mastitis,three lactating cows with subclinical mastitis(determined by using the standard California mastitis test,CMT) were assigned to the tested group,and three healthy lactating cows(characterized by udder redness,swelling,hardness,pain and grossly abnormal milk) were set to be control.Each experimental animal was treated with 1 600 μg(400 μg each quarter) of the mammary-specific expression vector p205C3LYZ described elsewhere,used the 5′-and 3...