A practical guide to Nosema qPCR

Honeybees face mounting environmental pressures, including pesticides, shrinking habitats, and a changing climate. Amid these threats lies Nosema ceranae, a microscopic gut parasite that saps bees’ energy and shortens their lives. How can we detect this pathogen accurately and early? Traditionally, researchers have relied on spore counts viewed under a microscope. While useful, this method struggles when infections are light or just beginning. To gain a more sensitive picture of infection, we can turn to molecular techniques (qPCR). However, many qPCR protocols described in the literature are incomplete, ambiguous, or missing critical practical details. Different labs often must “fill in the blanks,” leading to inconsistent results and reduced reproducibility.

Standard amplification curves, as produced with the protocol by Antoł & Ostap-Chec. Figure by Weronika Antoł.

A detailed protocol published by Weronika and Monika helps to bridge that gap. It provides a comprehensive, step-by-step guide for quantifying Nosema ceranae in individual bees using qPCR, focusing on clarity, transparency, and reproducibility. The method targets the parasite’s Hsp70 gene. As each spore contains exactly one copy, gene counts translate directly into spore numbers. The protocol also includes practical guidance often missing elsewhere, from homogenizing bee abdomens to preparing standards and troubleshooting.

A useful resource that we hope will facilitate the use of this technique in research and practice. Check it out on the protocols.io platform.