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

J Clin Microbiol

CBR Division, Defence Science and Technology Laboratory (Dstl), Porton Down, Salisbury, Wiltshire, United Kingdom.

Published: October 2015

AI Article Synopsis

  • Rapid inactivation of the Ebola virus (EBOV) is essential for effective testing in low-resource outbreak situations.
  • Testing revealed that Buffer AVL (commonly used for RNA extraction) was ineffective in inactivating EBOV in 67% of samples, indicating it shouldn't be relied upon for virus inactivation in diagnostics.
  • However, combining Buffer AVL with ethanol or heat treatments resulted in complete viral inactivation in all tested samples, while still allowing for the extraction of high-quality RNA for PCR analysis.

Article Abstract

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.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4572529PMC
http://dx.doi.org/10.1128/JCM.01449-15DOI Listing

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