用于阿龙山病毒检测的定量聚合酶链反应系统

Quantitative Polymerase Chain Reaction System for Alongshan Virus Detection.

作者信息

Litov Alexander G, Okhezin Egor V, Kholodilov Ivan S, Polienko Alexandra E, Karganova Galina G

机构信息

Laboratory of Biology of Arboviruses, FSASI "Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of RAS" (Institute of Poliomyelitis), 108819 Moscow, Russia.

Department of Biology, Lomonosov Moscow State University, 119234 Moscow, Russia.

出版信息

Methods Protoc. 2023 Sep 4;6(5):79. doi: 10.3390/mps6050079.

DOI:10.3390/mps6050079

PMID:37736962

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10514782/

Abstract

The recently discovered Jingmenvirus group includes viruses with a segmented genome, RNA of a positive polarity, and several proteins with distant homology to the proteins of the members of the genus Some Jingmenvirus group members, namely the Alongshan virus (ALSV) and Jingmen tick virus, are reported to be tick-borne human pathogens that can cause a wide variety of symptoms. The ALSV is widely distributed in Eurasia, yet no reliable assay that can detect it exists. We describe a qPCR system for ALSV detection. Our data showed that this system can detect as little as 10 copies of the ALSV in a sample. The system showed no amplification of the common tick-borne viruses circulating in Eurasia, i.e., the Yanggou tick virus-which is another Jingmenvirus group member-or some known members of the genus . The qPCR system was tested and had no nonspecific signal for the , , , and ticks. The qPCR system had no nonspecific signal for human and sheep serum as well. Overall, the qPCR system described here can be used for reliable and quantitative ALSV detection.

摘要

最近发现的荆门病毒群包括基因组呈节段状、RNA为正链以及几种与该属成员的蛋白质具有远缘同源性的蛋白质的病毒。据报道，荆门病毒群的一些成员，即阿龙山病毒（ALSV）和荆门蜱病毒，是蜱传播的人类病原体，可引起多种症状。ALSV广泛分布于欧亚大陆，但目前尚无可靠的检测方法。我们描述了一种用于检测ALSV的qPCR系统。我们的数据表明，该系统能够检测到样本中低至10个拷贝的ALSV。该系统对欧亚大陆流行的常见蜱传病毒，即另一种荆门病毒群成员——杨沟蜱病毒或该属的一些已知成员，均无扩增反应。该qPCR系统在对长角血蜱、全沟硬蜱、嗜群血蜱和微小牛蜱进行检测时未出现非特异性信号。该qPCR系统对人血清和羊血清也无非特异性信号。总体而言，本文所述的qPCR系统可用于可靠且定量的ALSV检测。