梯型熔解温度等温扩增检测食品中植物源成分的内标方法构建
目的 为梯型熔解温度核酸等温扩增(ladder-type melting temperature isothermal amplification,LMTIA)检测食品中植物源特异性基因建立内标方法,并用于检测肉制品中植物成分.方法 针对植物转录间隔区(internal transcribed spacer,ITS)的共有序列,设计LMTIA引物,优化反应温度,考查方法灵敏度,并评估淀粉制品、蜂蜜、食用油DNA提取方法与LMTIA检测方法的适应性,以H2O、鸭肉、鸡肉、羊肉、猪肉和牛肉基因组为阴性对照测试LMTIA方法的假阳性.结果 所建立的LMTIA方法在最优温度57 ℃时,灵敏度为10 p...
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| Published in | 食品安全质量检测学报 Vol. 12; no. 21; pp. 8439 - 8445 |
|---|---|
| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
许昌学院河南省食品安全生物标识快检技术重点实验室,许昌461000%河南科技大学食品与生物工程学院,洛阳471000
2021
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| Subjects | |
| Online Access | Get full text |
| ISSN | 2095-0381 |
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| Abstract | 目的 为梯型熔解温度核酸等温扩增(ladder-type melting temperature isothermal amplification,LMTIA)检测食品中植物源特异性基因建立内标方法,并用于检测肉制品中植物成分.方法 针对植物转录间隔区(internal transcribed spacer,ITS)的共有序列,设计LMTIA引物,优化反应温度,考查方法灵敏度,并评估淀粉制品、蜂蜜、食用油DNA提取方法与LMTIA检测方法的适应性,以H2O、鸭肉、鸡肉、羊肉、猪肉和牛肉基因组为阴性对照测试LMTIA方法的假阳性.结果 所建立的LMTIA方法在最优温度57 ℃时,灵敏度为10 pg/μL玉米基因组与10 pg/μL红枣基因组,可检出肉制品掺入0.1％的玉米淀粉,所选淀粉制品、蜂蜜、食用油DNA提取方法均适用于LMTIA检测,并且所有阴性对照样品在测试中均未出现假阳性.结论 本研究成功建立了LMTIA检测食品中植物源特异性基因的内标方法,排除了DNA提取引起的LMTIA检测的假阴性,可用于肉制品中植物成分定性检测. |
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| AbstractList | 目的 为梯型熔解温度核酸等温扩增(ladder-type melting temperature isothermal amplification,LMTIA)检测食品中植物源特异性基因建立内标方法,并用于检测肉制品中植物成分.方法 针对植物转录间隔区(internal transcribed spacer,ITS)的共有序列,设计LMTIA引物,优化反应温度,考查方法灵敏度,并评估淀粉制品、蜂蜜、食用油DNA提取方法与LMTIA检测方法的适应性,以H2O、鸭肉、鸡肉、羊肉、猪肉和牛肉基因组为阴性对照测试LMTIA方法的假阳性.结果 所建立的LMTIA方法在最优温度57 ℃时,灵敏度为10 pg/μL玉米基因组与10 pg/μL红枣基因组,可检出肉制品掺入0.1％的玉米淀粉,所选淀粉制品、蜂蜜、食用油DNA提取方法均适用于LMTIA检测,并且所有阴性对照样品在测试中均未出现假阳性.结论 本研究成功建立了LMTIA检测食品中植物源特异性基因的内标方法,排除了DNA提取引起的LMTIA检测的假阴性,可用于肉制品中植物成分定性检测. |
| Author | 张萌 郭孝辉 肖付刚 孙军涛 王德国 王永真 张永清 |
| AuthorAffiliation | 许昌学院河南省食品安全生物标识快检技术重点实验室,许昌461000%河南科技大学食品与生物工程学院,洛阳471000 |
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| Author_FL | WANG De-Guo GUO Xiao-Hui ZHANG Meng ZHANG Yong-Qing XIAO Fu-Gang WANG Yong-Zhen SUN Jun-Tao |
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| DocumentTitle_FL | Establishment of internal standard method for detection of plant-derived components in food with ladder-type melting temperature isothermal amplification |
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| Keywords | 梯型熔解温度等温扩增;植物源成分;特异性基因;内标方法;DNA提取方法 |
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| PublicationTitle | 食品安全质量检测学报 |
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| Publisher | 许昌学院河南省食品安全生物标识快检技术重点实验室,许昌461000%河南科技大学食品与生物工程学院,洛阳471000 |
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| Title | 梯型熔解温度等温扩增检测食品中植物源成分的内标方法构建 |
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