蝙蝠生物监测研究用 Pan-SYBR Green qPCR 检测方法的建立

Development of a Pan- SYBR Green qPCR Assay for Biosurveillance Studies in Bats.

机构信息

Centre for Emerging Zoonotic and Parasitic Diseases, National Institute for Communicable Diseases, A Division of the National Health Laboratory Service, Johannesburg 2131, South Africa.

Centre for Viral Zoonoses, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, Pretoria 0001, South Africa.

出版信息

Viruses. 2023 Apr 17;15(4):987. doi: 10.3390/v15040987.

DOI:10.3390/v15040987

PMID:37112966

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10145118/

Abstract

Recent studies have indicated that bats are hosts to diverse filoviruses. Currently, no pan-filovirus molecular assays are available that have been evaluated for the detection of all mammalian filoviruses. In this study, a two-step pan-filovirus SYBR Green real-time PCR assay targeting the nucleoprotein gene was developed for filovirus surveillance in bats. Synthetic constructs were designed as representatives of nine filovirus species and used to evaluate the assay. This assay detected all synthetic constructs included with an analytical sensitivity of 3-31.7 copies/reaction and was evaluated against the field collected samples. The assay's performance was similar to a previously published probe based assay for detecting Ebola- and Marburgvirus. The developed pan-filovirus SYBR Green assay will allow for more affordable and sensitive detection of mammalian filoviruses in bat samples.

摘要

最近的研究表明，蝙蝠是多种丝状病毒的宿主。目前，尚无经过评估可检测所有哺乳动物丝状病毒的通用丝状病毒分子检测方法。在这项研究中，开发了一种针对核蛋白基因的两步通用丝状病毒 SYBR Green 实时 PCR 检测方法，用于蝙蝠中的丝状病毒监测。合成构建体被设计为 9 种丝状病毒的代表，并用于评估该检测方法。该检测方法可检测到包含的所有合成构建体，分析灵敏度为 3-31.7 拷贝/反应，并针对现场采集的样本进行了评估。该检测方法的性能与之前发表的用于检测埃博拉病毒和马尔堡病毒的探针检测方法相似。开发的通用丝状病毒 SYBR Green 检测方法将能够更经济实惠和敏感地检测蝙蝠样本中的哺乳动物丝状病毒。