The European corn borer (Ostrinia nubilalis (Hübner)) is one of the most serious corn pest in Europe where it is controlled with pesticides, in particular, pyrethroids. First control failures with this chemical family occurred on the field in 2008 in the center of France, and the first resistance case was described in 2012. In the present study, we investigate resistance mechanisms involved in seven French populations of O. nubilalis collected in the field. Resistances to deltamethrin and lambda-cyhalothrin were confirmed, with a higher resistance ratio for lambda-cyhalothrin (63.79 compared to 7.67). Resistance to the two active compounds was correlated except for one population, indicating a high probability of cross-resistance. Analyses of the activity of three major families of detoxification enzymes in resistant individuals showed a significant increase of the average MFO activity in males of four populations (activity ratios of 2.76–5.73) and higher GST activity in females of two other populations (activity ratios 4.48 and 5.21). Molecular investigation of the sodium channel gene sequence showed the presence of the kdr mutation in a highly resistant individual. We designed a PCR-RFLP screening tool to search for this mutation in the field, and we found it in five populations but not in the susceptible one. The resistance of O. nubilalis to pyrethroids in France seems to result from a combination of resistance mechanisms, possibly as a consequence of a selection pressure with an exceptional duration (almost 40 yr old).