摘要
为评价奶山羊接种布鲁氏菌S2疫苗后全血和羊乳样本中疫苗菌体驻留情况及检样中提取DNA的最佳得率,分别采集布鲁氏菌抗体呈阳性和阴性的全血和羊乳样本各10份,提取全血及羊乳中的全基因组DNA,根据实验室建立的实时荧光定量PCR(qPCR)方法检测全血和羊乳中是否含有布鲁氏菌疫苗菌;分析全血和羊乳qPCR检测中Ct值差异,评价2种样本中的菌体载量。结果显示,从羊乳不同组分中提取全基因组DNA,以离心后沉淀中菌体核酸含量最高。在10只布鲁氏菌抗体呈阳性的全血样本中,均检测到S2疫苗菌;在10只布鲁氏菌抗体呈阳性的羊乳样本中,9份样本检测到布鲁氏菌S2疫苗株核酸信号,羊乳Ct值低于全血的Ct值。布鲁氏菌抗体呈阴性的全血和羊乳样本中,3份样本检测到S2疫苗株核酸信号。结果表明,该研究成功检测到奶山羊全血和羊乳中S2疫苗菌体驻留,试验结果可为奶山羊布鲁氏菌病的免疫防控提供参考。
This study aimed to evaluate the persistence of Brucella S2 vaccine strain in whole blood and milk samples from dairy goats after immunization and to determine the optimal DNA yield for bacterial detection.A total of 10 whole blood and 10 milk samples were collected from Brucella antibody-positive and antibody-negative goats,respectively,and whole genome DNA was extracted from these samples.Subsequently,a laboratory-established real-time quantitative PCR(qPCR)assay was employed to detect Brucella vaccine nucleic acids,and an additional comparison made based on cycle threshold(Ct)values was analyzed to evaluate the bacterial load for both sample types.The results revealed that whole genome DNA extracted from the precipatate after centrifugation of milk samples yielded the highest nucleic acid content.Brucella S2 vaccine nucleic acids were detected in all 10 Brucella antibody-positive whole blood samples and in 9 out of 10 antibody-positive milk samples.Moreover,Ct values obtained from milk samples were lower than those from whole blood.In the antibody-negative group,three samples(including both whole blood and milk)tested positive for Brucella S2 vaccine nucleic acids.These findings demonstrate successful identification of the Brucella S2 vaccine strain in both whole blood and milk of dairy goats using a valid clinical method.This study presents a practical approach for monitoring and controlling brucellosis in dairy goats.
作者
张雅婷
陈兰
陈昕迪
王腾宇
曹天宇
于宇
张慧忠
王文龙
ZHANG Ya-ting;CHEN Lan;CHEN Xin-di;WANG Teng-yu;CAO Tian-yu;YU Yu;ZHANG Hui-zhong;WANG Wen-long(Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease of Ministry of Agriculture/College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot,Inner Mongolia,010010,China;Ordos Animal Disease Prevention and Control Center,Ordos,Inner Mongolia,017000,China;Dongsheng District Agricultural and Livestock Product Quality and Safety Workstation,Ordos,Inner Mongolia,017000,China)
出处
《动物医学进展》
北大核心
2025年第9期46-51,共6页
Progress In Veterinary Medicine
基金
内蒙古自治区科技重大专项(2021ZD0023)。
