中国人兽共患病学报2026，Vol.42Issue(3)：269-275,7.DOI:10.3969/j.issn.1002-2694.2026.00.063
马尔堡病毒双重实时荧光定量PCR检测方法的建立
Establishment of a dual real-time fluorescence quantitative PCR method for detection of marburg virus
摘要
Abstract
This study developed a dual-fluorescence RT-PCR method for rapid detection of marburg virus(MARV),a filovirus that causes hemorrhagic fever.Five primer and probe sets were designed to target variable regions of the MNP and MGP genes.The as-say was optimized,and its performance was validated with national reference materials for MARV nucleic acid detection.For valida-tion,we assessed the compliance rates of positive and negative controls,the method's precision,and its limit of detection.Evaluation of 19 national reference samples confirmed the method's high accuracy,and 100％compliance for both positive and negative controls.All 6 positive controls exhibited consistent amplification in the FAM and VIC channels,corresponding to MNP and MGP,and all seven negative controls yielded negative results.The assay demonstrated a low detection limit below 1.4×10⁴ copies/mL for the MNP gene and 2.0×10⁴ copies/mL for the MGP gene.Precision testing indicated excellent reproducibility,with coefficients of variation be-low 5％for all channels.These findings indicated that the dual-fluorescence RT-PCR method is specific,sensitive,and reproducible,thus meeting national reference criteria.The assay is therefore suitable for deployment in grassroots disease control and healthcare set-tings for effective MARV surveillance and outbreak early warning.关键词
马尔堡病毒/国家参考品/实时定量荧光PCR法/丝状病毒科Key words
marburg virus/national reference material/real-time quantitative fluorescence PCR/Filoviridae分类
医药卫生引用本文复制引用
彭博,蔡传良,石晓路,陈诗婷,任玉冰,张娟,石大伟..马尔堡病毒双重实时荧光定量PCR检测方法的建立[J].中国人兽共患病学报,2026,42(3):269-275,7.基金项目
深圳市"医疗卫生三名工程"项目(No.SZSM202211023) （No.SZSM202211023）
深圳市医学研究专项资金(No.B2404002) （No.B2404002）
深圳市科技计划资助(No.SYSPG 20241211173921049). Sanming Project of Medicine in Shenzhen(No.SZSM202211023) （No.SYSPG 20241211173921049）
Shenzhen Medical Research Fund(No.B2404002) （No.B2404002）
Shenzhen Science and Technology Program(No.SYSPG20241211173921049).Peng Bo and Cai Chuanliang contributed equally to this work and are co-first authors. （No.SYSPG20241211173921049）
