| 双重PCR快速检测百日咳杆菌 |
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| 引用本文： | 马卓娅,王和平,陈小文,邓继岿,张民,郑跃杰. 双重PCR快速检测百日咳杆菌[J]. 实用诊断与治疗杂志, 2011, 25(2) |
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| 作者姓名： | 马卓娅 王和平 陈小文 邓继岿 张民 郑跃杰 |
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| 作者单位： | 1. 深圳市儿童医院呼吸科,广东省,518026
2. 深圳市儿童医院儿科研究所,广东省,518026 |
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| 基金项目： | 深圳市重点医学专科项目 |
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| 摘 要： | 目的:以百日咳毒素S1亚基启动子ptxA-Pr基因和插入序列IS481为目的基因,建立敏感、特异的双重PCR快速检测百日咳杆菌方法.方法:运用百日咳杆菌ptxA-Pr和IS481基因序列特异性引物,采用双重PCR技术同时扩增百日咳杆茵的特异性基因ptxA-Pr和IS481.通过构建目的质粒获得阳性对照,测序并与GenBank比对序列验证扩增产物.百日咳标准茵株DNA 10倍系列稀释为模板.采用此方法扩增双基因,检测此方法敏感性.扩增肺炎克雷伯茵、肺炎链球茵、铜绿假单胞茵、金黄色葡萄球菌、大肠杆菌及阳性样本DNA,检测此方法特异性.结果:百日咳杆菌标准株和阳性样本均能同时扩增ptxA-Pr和IS481目标序列,分别为191、145 bp.构建质粒后测序结果与GenBank对比一致.每个反应体系能检测到的标准菌株核酸最小量为1.65×10<'-2>ng.其他菌株检测未出现非特异性扩增.结论:双重PCR扩增百日咳杆菌ptxA-Pr和IS481基因的方法可用采特异快速的检测百日咳杆菌.
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| 关 键 词： | 双重PCR 百日咳杆菌 快速检测 |
Duplex PCR assay for detecting specific gene sequence from pertussis bacillus |
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| MA Zhuo-ya,WANG He-ping,CHEN Xiao-wen,DENG Ji-kui,ZHANG Min,ZHENG Yue-jie. Duplex PCR assay for detecting specific gene sequence from pertussis bacillus[J]. Journal of Practical Diagnosis and Therapy, 2011, 25(2) |
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| Authors: | MA Zhuo-ya WANG He-ping CHEN Xiao-wen DENG Ji-kui ZHANG Min ZHENG Yue-jie |
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| Affiliation: | MA Zhuo-ya1),WANG He-ping1),CHEN Xiao-wen2),DENG Ji-kui1),ZHANG Min2),ZHENG Yue-jie1) 1)Department of Pulmonary Medicine,Shenzhen Children Hospital,Shenzhen 518026,China,2) Institute of Pediatrics of Shenzhen Children Hospital |
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| Abstract: | Objective To establish a rapid assay with high sensitivity and specificity based on the sequence for pertussis toxin S1 promoter ptxA-Pr and insertion element sequence IS481.Methods Oligonucleotide primers targeting the ptxA-Pr and IS481 were used to amplify in the same reaction.The amplicons were inserted into the plasmid,sequenced and compared in the GenBank.The sensitivity of pertussis bacillus assay was determined by amplifying pertussis bacillus standard bacterial strain DNA at 10 dilutions.The five ot... |
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| Keywords: | Duplex PCR pertussis bacillus rapid assay |
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