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Parasitology Research

, Volume 110, Issue 4, pp 1403–1410 | Cite as

Comparative analysis of detection limits and specificity of molecular diagnostic markers for three pathogens (Microsporidia, Nosema spp.) in the key pollinators Apis mellifera and Bombus terrestris

  • Silvio ErlerEmail author
  • Stefanie Lommatzsch
  • H. Michael G. Lattorff
Original Paper

Abstract

Global pollinator decline has recently been discussed in the context of honey and bumble bee infections from various pathogens including viruses, bacteria, microsporidia and mites. The microsporidian pathogens Nosema apis, Nosema ceranae and Nosema bombi may in fact be major candidates contributing to this decline. Different molecular and non-molecular detection methods have been developed; however, a comparison, especially of the highly sensitive PCR based methods, is currently lacking. Here, we present the first comparative quantitative real-time PCR study of nine Nosema spp. primers within the framework of primer specificity and sensitivity. With the help of dilution series of defined numbers of spores, we reveal six primer pairs amplifying N. apis, six for N. bombi and four for N. ceranae. All appropriate primer pairs detected an amount of at least 104 spores, the majority of which were even as sensitive to detect such low amounts as 103 to ten spores. Species specificity of primers was observed for N. apis and N. bombi, but not for N. ceranae. Additionally, we did not find any significant correlation for the amplified fragments with PCR efficiency or the limit of detection. We discuss our findings on the background of false positive and negative results using quantitative real-time PCR. On the basis of these results, future research might be based on appropriate primer selection depending on the experimental needs. Primers may be selected on the basis of specificity or sensitivity. Pathogen species and load may be determined with higher precision enhancing all kinds of diagnostic studies.

Keywords

Colony Collapse Disorder Nosema Species Nosema Infection Pollinator Decline Theoretical Detection Limit 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

We thank the group of Paul Schmid-Hempel (ETH Zurich) for providing N. bombi-infected bumble bees and Denise Kleber for technical assistance within the lab. Financial support was granted by the Bundesministerium für Bildung und Forschung (BMBF) program FUGATO-Plus (FKZ: 0315126 to HMGL).

Supplementary material

436_2011_2640_MOESM1_ESM.pdf (516 kb)
ESM 1 (PDF 515 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Silvio Erler
    • 1
    Email author
  • Stefanie Lommatzsch
    • 1
  • H. Michael G. Lattorff
    • 1
  1. 1.Institut für Biologie, Molekulare ÖkologieMartin-Luther-Universität Halle-WittenbergHalle (Saale)Germany

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