Despite several psychometric advantages over the 50-item Autism Spectrum Quotient, an instrument used to measure autistic traits, the abridged AQ-28 and its cross-cultural validity have not been examined as extensively. Therefore, this study aimed to examine the factor structure and measurement invariance of the AQ-28 in 818 Dutch (M[subscript age] = 37.4, 581 females, 233 males, 4 others) and 437 Malaysian (M[subscript age] = 23.0, 328 females, 99 males, 10 others) participants from the general population. The hierarchical structure of the AQ-28 showed fair and good fit in Malaysia and in the Netherlands, respectively. A multi-group invariance analysis supported that the AQ-28 is cross-culturally invariant. Malaysians (M = 68.63, SD = 8.33) scored significantly higher than Dutch participants (M = 51.48, SD = 10.30) on the AQ-28 while gender was controlled for. While the measurement invariance suggests that the AQ-28 functions similarly in Malaysia and the Netherlands in terms of structure, exploratory analyses showed 11 items with differential item functioning. Hence, while the AQ-28 possesses a stable factor structure and appears to measure the same latent traits in Malaysia and the Netherlands, some items potentially display cultural bias which, in turn, might explain the differences in AQ scores.