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Evaluation of sampling technique and transport media for the diagnostics of adenoviral eye infections

Adenovirus sampling and transport

  • Laboratory Investigation
  • Published:
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Abstract

Background

Human adenoviruses (HAdV) may cause pharyngoconjunctival fever, follicular conjunctivitis or epidemic keratoconjunctivitis (EKC). Especially, outbreaks of the latter may lead to severe economic losses when preventive measures are implemented too late. Thus, a safe sampling method, proper specimen transport conditions and a fast and sensitive diagnostic technique is mandatory.

Methods

Two commercially available virus transport systems (VTS) were compared with two NaCl-moisturised sampling devices, one of which comprises Dacron-tipped plastic-shafted swabs and the other a cotton-tipped wood-shafted swab, available in most ophthalmologists’ offices. Downstream methods for specific detection of HAdV included direct immunofluorescence assay (IFA) of conjunctival swabs, virus isolation by cell culture and quantitative real-time polymerase chain reaction (qPCR). Furthermore, the influence of application of local anaesthetics prior to swabbing on subsequent detection of HAdV was investigated.

Results

Application of local anaesthetics had a positive influence on the amount of swabbed cells, thus increasing the chance of obtaining positive results by IFA. Neither isolation of HAdV by cell culture nor by qPCR was negatively influenced by this pretreatment. Surprisingly, both commercially available VTS performed significantly worse than the NaCl-moisturised swabs. This was shown with regard to virus recovery rates in cell culture as well as viral genome copy numbers in the qPCR.

Conclusions

Based on our results, the following recommendations are provided to improve sampling, transport and diagnostic techniques regarding conjunctival swabs for diagnosis of human adenovirus infection: (1) application of local anaesthetics, (2) NaCl-moisturised VTS for shipment of specimens, and (3) detection of HAdV by qPCR. The latter method proved to be superior to virus isolation by cell culture, including subsequent identification by IFA, because it is faster, more sensitive and allows simultaneous handling of a number of samples. Hence, countermeasures to prevent further virus spread in an outbreak situation can be implemented earlier, thus reducing the number of subsequent adenoviral infections.

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Acknowledgements

We acknowledge the excellent technical support of Heike Prabel, Aileen Lorber, Peter Klein, Rahime Terzioglu and Gudrun Zöller during the course of this study. We thank Dr. Albert Heim, German National Reference Laboratory for Adenoviruses, Hanover Medical School, Germany, for his helpful support in setting up the quantitative HAdV real-time PCR. The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the German Ministry of Defence or the German Government.

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Authors and Affiliations

  1. Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, 80937, Munich, Germany

    Roman Wölfel, Martin Pfeffer, Sandra Essbauer & Gerhard Dobler

  2. Bundeswehr Medical Specialist Service Section of Ophthalmology, Munich, Germany

    Sylke Nerkelun

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  1. Roman Wölfel
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  2. Martin Pfeffer
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  3. Sandra Essbauer
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  4. Sylke Nerkelun
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  5. Gerhard Dobler
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Correspondence to Roman Wölfel.

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Wölfel, R., Pfeffer, M., Essbauer, S. et al. Evaluation of sampling technique and transport media for the diagnostics of adenoviral eye infections. Graefe's Arch Clin Exp Ophthalmo 244, 1497–1504 (2006). https://doi.org/10.1007/s00417-006-0283-9

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  • DOI: https://doi.org/10.1007/s00417-006-0283-9

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