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J Med Microbiol. 2018 Sep;67(9):1321-1325. doi: 10.1099/jmm.0.000812. Epub 2018 Jul 27.

Effect of storage time and temperature on the detection of Pseudomonas aeruginosa, Acanthamoeba and Herpes Simplex Virus from corneal impression membranes.

Author information

1
1​St Paul's Eye Unit, Royal Liverpool University Hospital, Liverpool, UK.
2
2​Department of Eye and Vision Sciences, University of Liverpool, Liverpool, UK.
3
3​Department of Infection and Immunity, Liverpool Clinical Laboratories, Royal Liverpool University Hospital, Liverpool, UK.

Abstract

The effect of storage time and temperature on the recovery of pathogen DNA from polytetrafluorethylene (PTFE) was investigated. PTFE impression membranes were inoculated with Pseudomonas aeruginosa, Herpes Simplex Virus-1 (HSV-1) or Acanthamoeba and stored at -70 °C, -20 °C, +4 °C or +35 °C. PCR was performed on days 0, 1, 2, 3, 7 and months 1, 3 and 10 post-inoculation. We found no reduction in the DNA recovery of any of the studied microorganisms for the first 3 days of storage up to +35 °C. For HSV-1 and P. aeruginosa, storage for 3 months at +35 °C was associated with a significant reduction in DNA recovery (P<0.001), but not at +4 °C, -20 °C or -70 °C for 1 month for P. aeruginosa and for 10 months for HSV-1. Acanthamoeba DNA recovery was not affected by any storage parameters (P=0.203). These results will inform the investigation of microbial keratitis where access to microbiological testing is not readily available.

KEYWORDS:

Acanthamoeba; HSV-1; P. aeruginosa; corneal impression membrane; microbial keratitis

PMID:
30051806
DOI:
10.1099/jmm.0.000812
[Indexed for MEDLINE]

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