Yamagata Prefectural Okitama Livestock Hygiene Service Center, Nanyo, Japan.
Forestry and Fisheries Department, Yamagata Prefecture Agriculture, Yamagata, Japan.
Anim Sci J. 2022 Jan-Dec;93(1):e13763. doi: 10.1111/asj.13763.
The concentration of inosine 5'-monophosphate (IMP) in beef is an important factor contributing to beef palatability. A previous study suggested that single nucleotide polymorphisms (SNPs) in the ecto-5'-nucleotidase (NT5E) gene strongly affect the concentration of IMP under postmortem conditions by regulating NT5E enzymatic activity in beef. Genotyping of the NT5E gene is performed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) or real-time PCR assay. However, these conventional laboratory assays require large installed instruments. They also involve complicated procedures and are time-consuming. Here, the PCR primers and probes for the NT5E gene (rs42508588 SNP) were designed and synthesized, and we examined the rapid genotyping of the NT5E gene using a PicoGene PCR 1100 mobile PCR device. The results showed that this system enabled rapid amplification of each allele at approximately 19.4 s per cycle, with a total run time of 13 min 36 s. This device is portable and does not require a power supply, which facilitates its use not only in specific laboratories but also in meat production farms and distribution stages of beef.
肌苷 5'-单磷酸（IMP）在牛肉中的浓度是影响牛肉口感的一个重要因素。先前的研究表明，核苷酸外切酶 5'（NT5E）基因中的单核苷酸多态性（SNP）通过调节牛肉中 NT5E 酶活性，强烈影响死后条件下 IMP 的浓度。NT5E 基因的基因分型采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）或实时 PCR 分析。然而，这些传统的实验室检测需要大型安装仪器。它们还涉及复杂的程序，并且耗时。在这里，我们设计和合成了 NT5E 基因（rs42508588 SNP）的 PCR 引物和探针，并使用 PicoGene PCR 1100 移动 PCR 设备检查了 NT5E 基因的快速基因分型。结果表明，该系统能够以大约每循环 19.4s 的速度快速扩增每个等位基因，总运行时间为 13 分钟 36 秒。该设备是便携式的，不需要电源，这不仅方便在特定实验室使用，也方便在牛肉生产农场和分销阶段使用。
