ABSTRACT

Identification of meat and meat products is significant for economic, religious, or public health reasons. This study focused on the detection and differentiation of meat animal species. A total of sixty-four meat samples (10 samples of each like beef, buffen, chevon, and mutton; twenty-four samples of pork) were analyzed for these purposes. PCR-RFLP of 12S rRNA gene with four restriction enzymes was performed on individual meat samples as well as mixture of different meat. Species specific primers as well as sequencing of DNA were also used to detect and verify the RFLP results. PCR amplification yielded a 456bp fragments from 100% samples. AluI digested beef DNA into 359 and 97bp, chevon and mutton DNA into 246 and 210bp fragments and pork DNA into 290 and 166bp fragments. However, it cannot digest buffen DNA. On the other hand HhaI digested only buffen DNA into 247 and 209bp fragments but not others. ApoI restricts only mutton DNA and yielded 329 and 127bp fragments but not others. Nevertheless, BspTI yielded 323 and 133bp fragments only from chevon. Besides, mixed samples (1:1) of beef-buffen and chevon-mutton can be differentiated by AluI & HhaI and BspTI & ApoI, respectively. Chord analysis based on our sequence and respective sequences of studied animals from GenBank indicated that the cutting sites are conserved in these species of animals. Phylogenetically each of the species was clustered separately. This technique may be useful for meat animal species detection and differentiation either from individual meat and mixed meat samples.

摘要

出于经济、宗教或公共健康的原因，肉和肉制品的识别具有重要意义。本研究侧重于肉类动物物种的检测和区分。出于这些目的，总共分析了 64 个肉类样品（牛肉、牛肉、山羊和羊肉各 10 个样品；猪肉 24 个样品）。12S rRNA 基因与四种限制性内切酶的 PCR-RFLP 对单个肉类样品以及不同肉类的混合物进行。物种特异性引物以及 DNA 测序也用于检测和验证 RFLP 结果。PCR 扩增从 100% 样品中产生 456bp 片段。铝我将牛肉 DNA 消化为 359 和 97 bp，将山羊和羊肉 DNA 消化为 246 和 210 bp 片段，将猪肉 DNA 消化为 290 和 166 bp 片段。但是，它不能消化 buffen DNA。另一方面，Hha我只将 buffen DNA 消化成 247 和 209bp 片段，而不是其他片段。Apo I 仅限制羊肉 DNA 并产生 329 和 127bp 片段，但不限制其他片段。尽管如此，BspT I 仅从 chevon 中产生了 323 和 133bp 的片段。此外，牛-buffen和chevon-mutton的混合样品（1：1）可以通过Alu I＆Hha I和BspT I＆Apo进行区分我分别。基于我们的序列和来自 GenBank 的研究动物的相应序列的和弦分析表明，切割位点在这些动物物种中是保守的。在系统发育上，每个物种被分别聚类。该技术可用于从单个肉类和混合肉类样品中检测和区分肉类动物物种。