Laboratory guidance and resources

The recommended sample type for testing BDBV is whole blood or plasma for patients, and oral swab for deceased individuals. Sampling from living or deceased patients suspected of Orthoebolavirus infection should only be performed by trained personnel using appropriate personal protective equipment. 

PUIs that test negative on blood specimens drawn less than 72 hours after symptom onset should remain in isolation and a second test should be performed on a fresh sample taken more than 72 hours following onset of symptoms. WHO guidance for diagnostics should be applied (Diagnostic testing for Ebola and Marburg virus diseases: interim guidance, 20 December 2024).

Patient management and medical needs may require additional testing besides molecular testing for BDBV such as differential diagnostics for Malaria and other diseases or blood culture etc. ECDC underlines the importance of handling samples from PUIs, probable and confirmed cases with appropriate biosafety measures according to a risk-based approach to perform sampling and any laboratory manipulation in a safe manner. Validated inactivation methods should be applied before testing is carried out. Inactivation and handling of specimens from PUI, probable or confirmed cases require a BSL 3 laboratory facility, or ─ after proper risk assessment ─ a BSL 2 laboratory facility using a Class III biosafety cabinet. WHO provides guidance on inactivation methods for different testing purposes (General Procedures for Inactivation of potentially infectious Samples with Ebola Virus - PAHO/WHO | Pan American Health Organization) as well as for the handling of samples from suspected Ebola disease (EBOD) cases . Following a risk-based approach it should be considered to draw only the minimum number of blood samples necessary as each additional tube increases exposure risk. Where possible testing for differential diagnoses should be carried out on inactivated samples using molecular testing (RT-PCR) to further reduce risks of infection. 

Where non-inactivated samples are required, appropriate risk management needs to be considered in consultation with IPC professionals, as well as microbiologists to establish correct risk management procedures in due time.  Using laboratory tests that can be carried out within the isolation unit including point of care tests/equipment can be options to carry out differential diagnosis or other required clinical testing on non-inactivated clinical samples. 

Laboratories are encouraged to verify that the tests used can detect BDBV and are not limited to Ebola virus (Orthoebolavirus zairense) or other orthoebolaviruses only. NAATs developed in-house should be verified against representative sequences from the current outbreak (e.g. see Ebola Bundibugyo - Browse | Pathoplexus). 

If the NAAT used does not distinguish between different orthoebolaviruses, positive findings should be confirmed by sequencing and/or by further testing using assays specific for BDBV. 

Further useful information on safe sample collection, sample handling and diagnostics is available in the following documents: 

• WHO interim guidance: How to safely collect blood samples by phlebotomy from patients suspected to be infected with Ebola or Marburg 
• WHO rapid advice guideline: How to safely collect oral swabs from deceased patients suspected to be infected with Ebola or Marburg 
• Diagnostic testing for Ebola and Marburg virus diseases: interim guidance, 20 December 2024
• General Procedures for Inactivation of potentially infectious Samples with Ebola Virus - PAHO/WHO | Pan American Health Organization
• Algorithm for handling of samples from suspected Ebola Virus Disease (EVD) - PAHO/WHO | Pan American Health Organization
• Safe use of personal protective equipment in the treatment of infectious diseases of high consequence

Handling of samples from PUIs, probable or confirmed cases should be conducted with appropriate biosafety measures according to a risk-based approach. Each laboratory should perform their own risk assessment based on the clinical specimens to be handled in combination with the procedures to be performed. National guidelines on laboratory biosafety should be followed. Inactivation and handling of specimens from PUIs, suspected or confirmed cases  require at least a BSL 3 laboratory facility, or ─ after proper risk assessment ─ BSL 2 laboratory facility using a Class III biosafety cabinet. Validated virus inactivation methods must be used before downstream testing of samples that may contain orthoebolaviruses. Good microbiological practice and procedure (GMPP) must be applied, including general behaviours and aseptic techniques, in line with WHO laboratory biosafety manual  and national provisions.  

Biosecurity requirements need to be fulfilled by appropriate risk management and laboratory safety procedures, having secure facilities, controlled access, competent personnel, and full specimen traceability, ensuring that biological materials are protected from misuse, loss, or unauthorised access. 

Links to WHO guidelines: 

• WHO Laboratory biosafety manual 
• WHO Laboratory biosecurity guidance

WHO’s guidance on regulations for the transport of infectious substances emphasises that all infectious samples must be classified correctly and transported in compliance with international regulations to prevent exposure during handling and transport. Patient specimens suspected of containing viable orthoebolaviruses, including BDBV, must be treated as Category A (UN2814) infectious substances. Proper labelling, documentation, and trained personnel are required to handle and ship the materials and clear coordination with the receiving laboratory must be established in advance. The overall aim is to ensure safe, traceable, and secure transport while minimising risks to transport workers, laboratory staff, and the public. 

Links to WHO guidelines:  

WHO guideline: Guidance on regulations for the transport of infectious substances, 2023-2024: applicable as from 1 October 2023  

WHO interim guidance: How to safely ship human blood samples from suspected Ebola or Marburg cases within a country by road, rail and sea 

The EURL-PH-ERZV supports Member States’ laboratories during the current Ebola disease outbreak by providing up‑to‑date diagnostic protocols, recommendations on appropriate assays for testing PUIs, probable or confirmed cases, and technical advice on biosafety, biosecurity, and laboratory procedures. It also facilitates coordination and information exchange across the EU laboratory network to support the response. 

Key activities include: 

• Diagnostic support, including primary and confirmatory testing of BDBV; 
• Provision of guidance and protocols for molecular detection of BDBV and other orthoebolaviruses; 
• Access to protocols and reagents for in-house assays used within the EURL consortium laboratories; 
• Provide information on available commercial and in-house diagnostic tests;  
• Advice on biosafety and biorisk management for handling suspected BDBV samples. 
• BDBV laboratory PCR protocol

For further information, please contact the EURL-PH-ERZV at: EURL-PH-ERZVfolkhalsomyndigheten [dot] se (EURL-PH-ERZV[at]folkhalsomyndigheten[dot]se)