| 贺乐1, 2，黄惠琴2*，朱军2，刘敏2，孙前光2，鲍时翔2.穿刺巴斯德芽菌荧光定量PCR检测方法的建立与应用[J].植物保护,2013,39(1):104-109. |
| 穿刺巴斯德芽菌荧光定量PCR检测方法的建立与应用 |
| Development and application of a real time PCR system for detection of Pasteuria penetrans |
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| DOI： |
| 中文关键词: 穿刺巴斯德芽菌 实时荧光定量PCR 定量检测 |
| 英文关键词:Pasteuria penetrans real time PCR quantitative detection |
| 基金项目: |
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| 摘要点击次数: 748 |
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| 中文摘要: |
| 以穿刺巴斯德芽菌16S rDNA片段为靶标，通过对荧光定量PCR反应条件的摸索，建立该菌的荧光定量PCR检测方法。所选靶点最适退火温度60 ℃，正、反向引物的最佳浓度搭配为900、300 nmol/L；以Ct值和质粒拷贝浓度对数为坐标轴建立标准曲线，回归方程为y=-3.200×logx+34.43，R2值为0.998，PCR扩增效率为1054%，对含穿刺巴斯德芽菌芽胞的土壤样品检测阈值为2×103个/g土壤。该方法敏感度高、特异性好，能够运用于穿刺巴斯德芽菌的定性、定量检测。 |
| 英文摘要: |
| Based on the 16S rDNA sequence, a real time PCR detection method for Pasteuria penetrans was developed. Its optimal annealing temperature was 60℃ and the most effective concentration of forward primer and reverse primer was 900 nmol/L and 300 nmol/L,respectively. There was a good linear relationship between log scaled copies of the plasmid and Ct values, and the regression equation was y=-3.200×logx+34.43, R2 value was 0.998, and reaction efficiency was 105.4%. The threshold for P. penetrans detection from soil sample was 2×103 spores/g soil. This system is considered to be effective and specific to be used for qualitative and quantitative detection of P. penetrans. |
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