Highly sensitive detection of pathogens is effective for screening meat during quarantine inspection and export. The “micro-amount of virion enrichment technique” (MiVET) was recently developed, which is a new method combining virus concentration with immunomagnetic beads and simple RNA extraction with sodium dodecyl benzenesulfonate (SDBS) for the specific and sensitive detection of avian influenza viruses (AIVs). AIV subtypes H3N2 and H4N2 were used to spike the surface of chicken breast meat samples. The modified MiVET protocol was tested by comparing it against three different homogenate preparation conditions, as well as in samples with added α-amylase and collagenase to digest inhibitors. The performance of the modified MiVET was evaluated by real-time RT-PCR assay targeting the matrix gene. Compared with conventional RNA extraction, the modified MiVET reproducibly concentrated AIVs in chicken meat samples with 100–1000-fold improvement by 60 s-hand homogenization. The 30 s- and 60 s-stomacher homogenizations resulted 100-fold and 10–100-fold improvement, respectively. The modified MiVET required < 60 min from homogenate preparation to final RNA elution. Further, use of the modified MiVET also decreased the rate of false-negative results. The modified MiVET is effective for the rapid and highly sensitive detection of AIVs in chicken meat samples, and can be applied to quarantine and export inspection at airports and seaports.

中文翻译：

---

改进的微量病毒体富集技术（MiVET）在检测鸡肉样品中的甲型禽流感病毒中的应用。

对病原体的高度灵敏检测可有效隔离检疫和出口肉。最近开发了“微量病毒粒子富集技术”（MiVET），这是一种将病毒浓度与免疫磁珠结合并通过十二烷基苯磺酸钠（SDBS）进行简单RNA提取的新方法，用于特异性和灵敏地检测禽流感病毒（ AIVs）。使用AIV亚型H3N2和H4N2对鸡胸肉样品的表面进行加标处理。通过将改良的MiVET方案与三种不同的匀浆制备条件进行比较，并在添加了α-淀粉酶和胶原酶以消化抑制剂的样品中进行了比较，测试了改良的MiVET方案。通过针对基质基因的实时RT-PCR分析评估了修饰的MiVET的性能。与传统的RNA提取相比，经过修饰的MiVET可重复浓缩的鸡肉样品中的AIV，经过60 s手工均质处理可提高100-1000倍。30 s和60 s气孔均质化分别提高了100倍和10-100倍。从匀浆制备到最终RNA洗脱，修饰的MiVET需<60分钟。此外，使用改良的MiVET还可以降低假阴性结果的发生率。改良的MiVET可有效快速，灵敏地检测鸡肉样品中的AIV，并可用于机场和港口的检疫和出口检验。从匀浆制备到最终RNA洗脱，修饰的MiVET需<60分钟。此外，使用改良的MiVET还可以降低假阴性结果的发生率。改良的MiVET可有效快速，灵敏地检测鸡肉样品中的AIV，并可用于机场和港口的检疫和出口检验。从匀浆制备到最终RNA洗脱，修饰的MiVET需<60分钟。此外，使用改良的MiVET还可以降低假阴性结果的发生率。改良的MiVET可有效快速，灵敏地检测鸡肉样品中的AIV，并可用于机场和港口的检疫和出口检验。