Nsawotebba Andrew, Nabadda Susan, Nakintu Valeria, Ssewanyana Isaac, Wayengera Misaki, Kabazzi Jonathan, Balinandi Stephen, Pimundu Godfrey, Muyigi Tonny, Ayitewala Alisen, Gidudu Samuel, Otita Morgan, Mugerwa Ibrahim, Rubangakene Moses, Ikoba Sulaiman, Kalungi Sam, Wanyenze Rhoda, Ariën Kevin K, Namboozo Eunice, Kanamwanji Benedict, Ssekyondwa Steven, Kasujja Ronald, Kitwe Derrick, Dambya Catherine, Morunyanga Innocent, Bahatungire Reginald Rony, Kakooza Francis, Joloba Moses, Kagirita Atek, Muruta Allan, Kaleebu Pontiano, Kyobe Henry, Olaro Charles, Atwine Diana, Aceng Jane Ruth
Department of National Health Laboratory and Diagnostic Services, Ministry of Health, Kampala, Uganda.
Central Emergency Response and Surveillance Laboratory, Department of National Health Laboratory and Diagnostic Services, Plot 106 - 1062, Old Butabika Road, P.O. Box 7272, Kampala, Uganda.
BMC Infect Dis. 2025 Jul 2;25(1):888. doi: 10.1186/s12879-025-11271-0.
On January 30, 2025, the Uganda Ministry of Health declared an outbreak of Sudan virus disease (SVD) following laboratory confirmation from postmortem samples of a suspected case identified through routine mortality surveillance for viral hemorrhagic fevers (VHFs) at Mulago National Referral Hospital, Kampala. This report describes the laboratory procedures used to confirm the 2025 SVD index case, emphasizing the vital role of rapid diagnostics in containing VHFs.
We leveraged existing surveillance infrastructure to collect, package, transport, test, and report results for the index-confirmed case. Testing was conducted within the framework of the established laboratory quality and bio-risk management system at the Central Emergency Response and Surveillance Laboratory. The workflow consisted of sample delivery, reception, preparation, nucleic acid extraction, master mix preparation, quantitative reverse transcriptase polymerase chain Reaction (RT-qPCR), differential diagnosis for VHFs, parallel PCR testing and typing for Sudan virus (SUDV) confirmation, result validation, and reporting.
The sample tested negative for Marburg, Yellow fever, Rift Valley fever, and Crimean-Congo Hemorrhagic Fever viruses. However, it tested positive for SUDV. Four parallel tests utilizing the Real Star® Filovirus Screen RT-PCR Kit 1.0 (Altona Diagnostics GmbH, Hamburg, Germany) on CFX96 Bio-Rad platforms, with varying sample volumes (140 µL, 90 µL, 50 µL, and 30 µL diluted with nuclease-free water), confirmed the presence of SUDV. The 50 µL sample had the lowest Ct values for the Orthoebolavirus target, while the 30 µL sample had the lowest Ct values for the internal control target. The turnaround time, from sample reception to results reporting to the Ministry of Health leadership, was less than 12 h.
Mortality surveillance of VHFs is vital for the swift identification of Filovirus outbreaks in high-risk areas. Comprehensive VHFs panel testing is crucial for detecting pathogens in suspected cases. Sample dilution impacts diagnostic sensitivity, making optimized testing protocols crucial for precise molecular diagnostics. A rapid turnaround time is critical to outbreaks, enabling timely public health decisions such as case isolation, contact tracing, and resource allocation.
2025年1月30日，乌干达卫生部在坎帕拉穆拉戈国家转诊医院对通过病毒性出血热（VHF）常规死亡监测发现的一例疑似病例进行尸检样本实验室确认后，宣布苏丹病毒病（SVD）暴发。本报告描述了用于确认2025年SVD索引病例的实验室程序，强调了快速诊断在控制VHF中的关键作用。
我们利用现有的监测基础设施收集、包装、运输、检测索引确认病例并报告结果。检测在中央应急响应和监测实验室既定的实验室质量和生物风险管理系统框架内进行。工作流程包括样本递送、接收、制备、核酸提取、主混合物制备、定量逆转录聚合酶链反应（RT-qPCR）、VHF鉴别诊断、苏丹病毒（SUDV）确认的平行PCR检测和分型、结果验证及报告。
该样本对马尔堡病毒、黄热病病毒、裂谷热病毒和克里米亚-刚果出血热病毒检测呈阴性。然而，对SUDV检测呈阳性。在CFX96 Bio-Rad平台上使用Real Star®丝状病毒筛查RT-PCR试剂盒1.0（德国汉堡Altona Diagnostics GmbH公司）进行了四项平行检测，样本体积不同（分别用无核酸酶水稀释至140µL、90µL、50µL和30µL），确认存在SUDV。50µL样本的正ebolavirus靶标的Ct值最低，而30µL样本的内控靶标的Ct值最低。从样本接收到向卫生部领导层报告结果的周转时间不到12小时。
VHF的死亡监测对于在高风险地区迅速识别丝状病毒暴发至关重要。全面的VHF检测组检测对于检测疑似病例中的病原体至关重要。样本稀释会影响诊断敏感性，因此优化检测方案对于精确的分子诊断至关重要。快速周转时间对疫情暴发至关重要，能够及时做出病例隔离、接触者追踪和资源分配等公共卫生决策。
Zotero 插件