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요약정보
1种布鲁氏菌微滴式数字PCR检测方法的建立 / A Droplet Digital PCR Method for Detection of Brucella
Resource Type
Academic Journal
Authors
梅力
;
王英超
;
程汝佳
;
于国际
;
范学政
;
高晓龙
;
高敏
;
秦玉明
;
李筱英
;
李巧玲
;
朱良全
;
冯小宇
;
MEI Li
;
WANG Yingchao
;
CHENG Rujia
;
YU Guoji
;
FAN Xuezheng
;
GAO Xiaolong
;
GAO Min
;
QIN Yuming
;
LI Xiaoying
;
LI Qiaoling
;
ZHU Liangquan
;
FENG Xiaoyu
Source
畜牧兽医学报 / Acta Veterinaria Et Zootechnica Sinica
. 52(6):1753-1759
Subject
布鲁氏菌
数字PCR
检测方法
Language
Chinese
ISSN
0366-6964
Online Access
Find it @ DONGA
초록
本研究旨在构建1种布鲁氏菌(Brucella)微滴式数字PCR方法(droplet digital PCR,ddPCR).以布鲁氏菌BCSP31基因为靶基因,选取保守区域设计引物和探针,构建了布鲁氏菌微滴式数字PCR方法.然后优化了ddPCR反应中的引物、探针浓度及退火温度,并进行方法的灵敏度、特异性和重复性试验.结果 表明,ddPCR的最佳引物和探针浓度分别为900 nmol·L-1和250 nmol·L-1,最佳的退火温度为58℃.ddPCR的线性良好,最低检测下限为1.12 copies·μL-1,与大肠杆菌O:157菌株等其他4种细菌无交叉反应,而且批内重复性和批间重复性都较好.综上表明,本研究建立的ddPCR方法灵敏度高、特异性强,可对布鲁氏菌感染的临床样品进行定量检测.
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