Buffer AVL Alone Does Not Inactivate Ebola Virus in a Representative Clinical Sample Type

• Published in: Journal of clinical microbiology 2015-10, Vol.53 (10), p.3148-3154
• Main Authors: Smither, Sophie J, Weller, Simon A, Phelps, Amanda, Eastaugh, Lin, Ngugi, Sarah, O'Brien, Lyn M, Steward, Jackie, Lonsdale, Steve G, Lever, Mark S
• Format: Article
• Language: English
• Subjects: Animals; Blood - virology; Buffers; Callithrix; Disinfectants - pharmacology; Ebolavirus - drug effects; Ebolavirus - physiology; Ethanol; Mice; Microbial Viability - drug effects; Virology; Virus Inactivation - drug effects
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/JCM.01449-15

Rapid inactivation of Ebola virus (EBOV) is crucial for high-throughput testing of clinical samples in low-resource, outbreak scenarios. The EBOV inactivation efficacy of Buffer AVL (Qiagen) was tested against marmoset serum (EBOV concentration of 1 × 10(8) 50% tissue culture infective dose per milliliter [TCID50 · ml(-1)]) and murine blood (EBOV concentration of 1 × 10(7) TCID50 · ml(-1)) at 4:1 vol/vol buffer/sample ratios. Posttreatment cell culture and enzyme-linked immunosorbent assay (ELISA) analysis indicated that treatment with Buffer AVL did not inactivate EBOV in 67% of samples, indicating that Buffer AVL, which is designed for RNA extraction and not virus inactivation, cannot be guaranteed to inactivate EBOV in diagnostic samples. Murine blood samples treated with ethanol (4:1 [vol/vol] ethanol/sample) or heat (60°C for 15 min) also showed no viral inactivation in 67% or 100% of samples, respectively. However, combined Buffer AVL and ethanol or Buffer AVL and heat treatments showed total viral inactivation in 100% of samples tested. The Buffer AVL plus ethanol and Buffer AVL plus heat treatments were also shown not to affect the extraction of PCR quality RNA from EBOV-spiked murine blood samples.