A two-centre validation study of sterility test of corneal storage media with elimination of interfering antimicrobials in compliance with the European Pharmacopoeia

Abstract

The purpose of this study was to validate the sterility test of corneal culture and deswelling/transport media using a device for removal of antimicrobials before incubation in BACTEC™ automated system in two Italian Eye Banks. Corneal culture medium, TISSUE-C, and deswelling/transport medium, CARRY-C, were inoculated with 10–100 cfu of six European Pharmacopoeia (EP) reference strains and either treated with medical device RESEP for removal of antimicrobials (RESEP+ group) or left untreated (RESEP− group) before injection into the BACTEC Plus bottles. The same steps were repeated in the absence of inocula with tryptone soy broth samples as negative controls, and the inocula were also directly injected in the BACTEC™ bottles as growth controls. All the samples were incubated in BACTEC™ automated system for 7 days, and the time to detection of microbial growth was recorded automatically. At both the Eye Banks, in the RESEP+ groups, microbial growth was detected in 100% of samples. In the RESEP− group, the method sensitivity ranged from 66.7 ± 21.1 to 88.9 ± 6.4% for TISSUE-C samples while for CARRY-C samples the method sensitivity ranged from 94.5 ± 5.1 to 100%. The method specificity corresponded to 100% for all the groups at both Eye Banks. This two-centre validation study showed that the use of RESEP increased the sensitivity of sterility test using BACTEC™ automated system up to 100% and, consequently, allowed validation of the method for sterility testing of corneal storage and deswelling/transport media according to the EP requirements. The test could not be validated without the use of RESEP.

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Acknowledgements

We would like to thank Cristina Gianotti for the writing assistance. No external funding was received for this study.

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Correspondence to J. D’Amato Tóthová.

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J. D’Amato Tóthová and L. Giurgola are employed by the company Alchilife S.r.l., which was involved in the development of one medical device discussed in this article. R. Vignola, R. Mistò, F. Pateri and A. Pocobelli have no financial or proprietary interest in any material or method mentioned.

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Vignola, R., Mistò, R., Giurgola, L. et al. A two-centre validation study of sterility test of corneal storage media with elimination of interfering antimicrobials in compliance with the European Pharmacopoeia. Cell Tissue Bank 20, 275–285 (2019). https://doi.org/10.1007/s10561-019-09766-7

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Keywords

  • Corneal culture media
  • Antimicrobials removal neutralization
  • Eye banking
  • Microbiological testing
  • Corneal storage and transplantation
  • European Pharmacopoeia sterility test