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从急性性患者的样中对脊髓灰质炎病毒的直接检测协议的评估

Minami Kikuchi Ueno¹, Kouichi Kitamura¹, Yorihiro Nishimura¹

¹Department of Virology II, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan.

|October 28, 2023

概括

直接检测方法为急性性 (AFP) 病例的脊髓灰质炎病毒 (PV) 监测提供了更快的替代方案. 一种针对VP4-VP2的方法显示出更高的灵敏度,在所有测试样本中检测到PV.

科学领域:

病毒学病毒学
分子生物学分子生物学
公共卫生公共卫生

背景情况:

目前的脊髓灰质炎监测依赖于从急性软性 (AFP) 病例的便样本中分离病毒.
现有的方法在报告时间表,测试灵敏度和脊髓灰质炎病毒 (PV) 隔离物制方面面临挑战.
目前正在探索直接检测 (DD) 方法作为潜在的替代方案.

研究的目的:

评估以前报告的脊髓灰质炎病毒 (PV) 直接检测 (DD) 方法的灵敏度和可序列性.
优化直接检测协议,以增强光伏检测和测序.

主要方法:

评估了针对 PV 基因组的 VP1 或 VP4-VP2 区域的直接检测 (DD) 协议.
优化了针对VP1测序的整个囊区域的RT-PCR协议.
对桑格和纳米孔测序的评估灵敏度 (检测极限) 和可测序性.

主要成果:

一个优化的VP1目标协议显示了高灵敏度 (LOD=82副本) 和更快的反应时间 (2.5小时).
针对VP1的DD方法在60-80%的阳性AFP样本中检测到PV,混合样本中缺少小PV菌株.
针对VP4-VP2的DD方法显示了更高的灵敏度 (LOD=25个副本),并在所有测试样本中检测到PV.

结论:

直接检测 (DD) 方法在AFP监测中有效检测脊髓灰质炎病毒 (PV).
针对VP4-VP2的DDD方法显示,它是目前监控方法的敏感替代方案.
进一步提高DD方法灵敏度对于替代传统病毒隔离技术至关重要.

关键词:

全球脊髓灰质炎实验室网络直接检测可以直接检测.纳米孔测序的测序非脊髓灰质炎肠道病毒脊髓灰质炎病毒是什么?

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