马尔堡病毒双重实时荧光定量PCR检测方法的建立OA

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.