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当前位置: 首页 > 文章 > 快速检测炭疽杆菌的实时荧光定量PCR方法的建立及评价 中国动物传染病学报 2019,27 (4) 43-49
Position: Home > Articles > DEVELOPMENT OF REAL-TIME FLUORESCENCE QUANTITATIVE PCR FOR RAPID DETECTION OF BACILLUS ANTHRACIS Chinese Journal of Animal Infectious Diseases 2019,27 (4) 43-49

快速检测炭疽杆菌的实时荧光定量PCR方法的建立及评价

作  者：
杜清春;董珊珊;丁奕博;张艳;李伟;王鹏
单  位：
云南省地方病防治所 云南省自然疫源性疾病防控技术重点实验室;大理大学公共卫生学院;中国疾病预防控制中心传染病预防控制所
关键词：
炭疽杆菌;实时荧光定量PCR;TJHP;BA5345
摘  要：
为建立一种能快速鉴定炭疽杆菌的检测技术,本研究根据炭疽杆菌染色体上的TJHP和BA5345 2个靶点,分别设计4对引物和2个探针,对10份已知浓度的炭疽杆菌DNA和阴性对照菌株DNA进行检测,将浓度最大的炭疽杆菌DNA进行10倍倍比稀释,用于检测4对引物及其探针的灵敏度,并选出2个针对TJHP和BA5345靶点的特异性强及灵敏度高的引物和探针进行重复性实验。结果显示,4对引物及其探针特异性较强,FP2/RP2和FP4/RP4 2对引物的灵敏度为4.513×10-5 ng/μL;该荧光定量PCR方法重复性好(t=1.178,P=0.332)。FP2/RP2和FP4/RP4 2对引物及其探针可用于炭疽杆菌感染初期的快速筛查以及临床辅助诊断。
译  名：
DEVELOPMENT OF REAL-TIME FLUORESCENCE QUANTITATIVE PCR FOR RAPID DETECTION OF BACILLUS ANTHRACIS
作  者：
DU Qing-chun;DONG Shan-shan;DING Yi-bo;ZHANG Yan;LI Wei;WANG Peng;The School of Public Health of Dali University;The Yunnan Key Laboratory of Zoonosis Control and Prevention,Yunnan Institute for Endemic Diseases Control and Prevention;National Institute of Infectious Diseases Control and Prevention, Chinese Center for Diseases Control and Prevention;
关键词：
Bacillus anthracis;;real-time fluorescence quantitative PCR;;TJHP;;BA5345
摘  要：
To develop a rapid detection method for Bacillus anthracis(B.anthracis), four pairs of primers and probes were designed according two target genes TJHP and BA5345 on the chromosomes. Then, these primers and probes were used to detect 10 known dilutions of B. anthracis samples for assay optimization. As a result, two pairs of primers and probes that had high specificity and sensitivity were selected for rapid detection of B. anthracis. The detection limit was 4.513×10-5 ng/uL for both FP2/RP2 and FP4/RP4 primers. The PCR assay developed here also showed good repeatability(t=1.178, P=0.332). These results showed that this real-time fluorescence quantitative PCR assay was suitable for rapid detection of B. anthracis in early infection and clinical diagnosis.

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