| 龙 阳1，马新华1，袁俊杰1，卢乃会1，杨卓瑜1，魏 霜2，王卫芳2.油菜茎基溃疡病菌LAMP-HNB 检测方法的建立[J].广东农业科学,2017,44(10):66-69 |
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油菜茎基溃疡病菌LAMP-HNB 检测方法的建立 |
| Establishment of LAMP-HNB method for detection of Leptosphaeria maculans in rape |
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| DOI：10.16768/j.issn.1004-874X.2017.10.011 |
| 中文关键词: 油菜 茎基溃疡病菌 环介导等温扩增 羟基萘酚蓝 检疫 |
| 英文关键词: rape Leptosphaeria maculans LAMP HNB quarantine |
| 基金项目:广东出入境检验检疫局科技计划项目（2016GDK32） |
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| 摘要点击次数: 2311 |
| 全文下载次数: 920 |
| 中文摘要: |
| 油菜茎基溃疡病菌是油菜上最严重的真菌病害之一，是我国重要的检疫性有害生物。因此为
防范其危害我国油菜产业生产安全，建立油菜茎基溃疡病菌的快速筛查方法具有重要意义。基于环介导等
温扩增法（LAMP） ，以ITS（内转录间隔区）为靶序列并结合指示剂羟基萘酚蓝（HNB）建立了一种基于
颜色判定的简单、快速和灵敏的油菜茎基溃疡病菌的快速检测方法，并对其特异性、灵敏性进行评价以及
实际样品进行检测。结果显示，该方法在62℃等温条件下仅需80 min 即可通过肉眼直接目测试验结果。
LAMP-HNB 法与标准PCR 检测方法对比显示其具有良好的特异性以及更高的灵敏度，利用该方法对10 批
油菜籽样品进行检测，共检出5 批阳性样品，与标准中PCR 检测方法结果一致。该方法的建立为油菜茎基
溃疡病菌的检疫鉴定提供了新途径。 |
| 英文摘要: |
| Leptosphaeria maculans is one of the most serious fungal diseases in rape. It is important to establish a
rapid detection method to protect the safety of rapeseed industry in China. The loop-mediated isothermal amplification
（LAMP） method was developed to detect L. maculans，and the hydroxy naphthol blue（HNB）was used as
indicator. The specificity and sensitivity of the method were tested. The result can be obtained within 80 minutes at
62℃ and judged by the eye. The results showed that LAMP-HNB method had a higher sensitivity than conventional
PCR. A total 5 positive samples for L. maculans were detected from 10 samples by the LAMP-HNB method，which
was in accordance with the result by conventional PCR. LAMP-HNB is a new method to detect L. maculans. |
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