不同线粒体DNA(mtDNA)、ND5、D-Loop和Cty-b DNA引物在检测猪DNA片段中的特异性

J. Kusnadi, N. Ashari, E. L. Arumingtyas
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引用次数: 2

摘要

聚合酶链反应(PCR)是一种准确、简便、快速的分析方法。该技术广泛应用于肉类掺假和肉类加工食品的鉴定。采用线粒体DNA (mt-DNA)引物NADH脱氢酶亚基5 (ND5)、D-Loop和细胞色素b (Cyt-b)检测猪(Sus scrofa) DNA片段的特异性。采用三对引物(ND5、D-Loop和Cyt-b)对6份肉类样品(猪肉、牛肉、山羊、羊肉和鸡肉)的DNA基因组进行PCR扩增和测序。三种引物扩增结果分别为232 bp、951 bp和404 bp。与ND5、D-Loop和Cyt-b基因序列比较,相似度分别为100%、97%和99%。结果表明,ND5、D-Loop和Cyt-b基因的mt-DNA引物可作为猪(苏氏)DNA片段检测的特异性引物。
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Specificity of Various Mitochondrial DNA (mtDNA), ND5, D-Loop, and Cty-b DNA Primers in Detecting Pig (Sus scrofa) DNA Fragments
Polymerase Chain Reaction (PCR) is an accurate, simple and fast analytical method. This technique is widely used in the identification of meat adulteration and meat-based processed food products. Three Mitochondrial DNA (mt-DNA) primers NADH Dehydrogenase sub unit 5 (ND5), D-Loop, and Cytochrome b (Cyt-b) were tested for their specificity in detecting of pig (Sus scrofa) DNA fragments. DNA genome from 6 meat samples (pork, beef, goat, lamb, and chicken) was amplified by PCR technique using three pairs of primers (ND5, D-Loop, and Cyt-b) and sequenced. The results of amplification using the three primers produced specific DNA bands with the lengths of 232 bp, 951 bp, and 404 bp, respectively. Comparison results with ND5, D-Loop, and Cyt-b gene sequences resulted in similarity values of 100%, 97%, and 99%, respectively. These showed that the mt-DNA primers of ND5, D-Loop, and Cyt-b genes can be recommended as specific primers in detecting pig (Sus scrofa) DNA fragments.
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