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Katrina L Counihan¹, Siddhartha Kanrar¹, Shannon Tilman¹

¹Eastern Regional Research Center, United States Department of Agriculture, Agricultural Research Service, Wyndmoor, PA 19038, USA.

Foods (Basel, Switzerland)

|March 28, 2024

概括

长时间读取的测序显示,可以很快地检测出在碎牛肉中产生Shiga毒素的细菌大肠杆菌 (STEC). 这种方法可以显著减少识别食物传播病原体所需的时间,提高食品安全.

科学领域:

食品微生物学食品微生物学
基因组学就是基因组学.
病原体检测检测病原体检测

背景情况:

食品传递的病原体,包括产生Shiga毒素的大肠杆菌 (STEC),对公共健康构成重大风险.
目前在肉类中使用的STEC检测方法耗时,通常需要四天以上.
长读全基因组测序为更快的病原体识别提供了潜在的途径.

研究的目的:

评估长读测序用于检测土牛肉中STEC的有效性.
优化测序参数并确定STEC病毒性基因的检测极限.
评估选择性测序和丰富策略,以加强STEC检测.

主要方法:

利用牛津纳米孔技术的MinION测序器进行长读全基因组测序.
优化了测序参数,包括1kb的最小读取长度.
确定了STEC病毒性基因 (eae,stx1,stx2,fliC,wzx,wzy,rrsC) 在纯种植物和有刺的碎牛肉中的检测极限.
评估选择性测序和在修改的三角豆 (mTSB) 中进行丰富.

主要成果:

所有目标STEC病毒性基因在测序后1小时内在纯种植物中被检测出来.
病毒性基因的高可靠性检测只用12.5 ng的STEC DNA来实现.
选择性测序改善了刺牛肉的检测,尽管发现了牛DNA干扰.
与未缩样本相比,mTSB的丰富导致病毒性基因检测增加了100倍.

结论:

长读测序是一种可行的方法,用于在纯种植中快速检测STEC.
需要进一步的协议开发来克服诸如牛DNA干扰在复杂的矩阵中的挑战,如碎牛肉.
优化的长读测序协议可以在食品安全应用中实现更快,更少的劳动密集型STEC检测.

关键词:

食品测试食品测试食品测试通过食物传播的病原体开发方法开发方法的方法.病毒性基因是病毒性基因.进行全基因组测序.

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