| 钟文涛 王芳妹 李白玉 蒋伟 周丛.四重PCR 反应体系的建立及在牛肉掺假检测中的应用[J].现代生物医学进展英文版,2016,16(18):3574-3577. |
| 四重PCR 反应体系的建立及在牛肉掺假检测中的应用 |
| Establishment of Quadruple PCR Reaction System: Its Application inDetection of Adulterated Beef |
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| DOI： |
| 中文关键词: 四重PCR 细胞色素b 牛肉掺假 检测 |
| 英文关键词: Quadruple PCR Cytochrome b Adulterated beef Detection |
| 基金项目:湖南省2012 年第六批省级科技计划项目（2012SK3290） |
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| 中文摘要: |
| 目的：建立马、牛、猪、鸭肉四重聚合酶链式反应（PCR）反应体系，为牛肉掺假检测提供参考。方法：选取马、牛、猪、鸭肉阳性
样本，根据线粒体基因组中细胞色素b（cytochrome b）序列，设计多重PCR引物，同时对马、牛、猪、鸭肉进行PCR及电泳检测，建
立四重PCR 反应体系，并运用该体系对市场40 份牛肉制品进行检测。结果：引物比例为马：牛：猪：鸭=1:1:2:1 时，46℃退火温度
下反应35 个循环，四重PCR效果较好；建立四重PCR反应体系，四重PCR 条带清晰且亮度较为一致，几乎没有二聚体产生，扩
增灵敏度高。根据样品检测结果，烤牛肉串、牛肉丸的掺假率较高，分别为36.4%、75%，牛排未检出除牛肉外的三种动物源成分，
但不排除有其他动物源成分的掺入。结论：四重PCR 反应体系可用于牛肉掺假检测。 |
| 英文摘要: |
| To establish the quadruple PCR reaction system of horsemeat, beef, pork and duck,in order to provide a
reference for the detection of adulterated beef. The positive samples of horsemeat, beef, pork and duck were selected. The
multiplex PCR primers were designed according to the sequence of cytochrome b in the mitochondrial genome; the horsemeat, beef, pork
and duck were detected by PCR and electrophoresis test at the same time; the quadruple PCR reaction system was established, then the
system was used to detect 40 beef products in the market. The effect of quadruple PCR was better when the ratios of primer of
horsemeat were beef: pork: duck=1:1:2:1, and 35 cycles reacted under the 46℃ annealing temperature. The quadruple PCR reaction
system was established, the strip was clear and bright, almost no dimer was generated, and the amplified sensitivity was high. The beef
products detection results showed that the adulteration rate of roast beef string,stewed beef meatballs were higher, which were 36.4%and
75%respectively, while the beefsteak had no other three kinds of animal ingredients except beef,but can not rule out the incorporation of
other animal ingredients. The quadruple PCR reaction systemcan be applied to detecting adulterated beef. |
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