Cancer has become a major killer threatening human health in today's society, and surface enhanced Raman spectroscopy (SERS) may provide a competitive choice for the sensitive detection of cancer‐related miRNAs. In this work, a synergistic calibrated SERS strategy based on MXene/molybdenum disulfide (MoS2)@Au nanoparticles (AuNPs) with controllable morphology is suggested for the ultrasensitive detection of miRNA‐182 by selecting the average intensity of its three own characteristic Raman peaks (at 382 cm−1 and 402 cm−1 corresponding to MoS2 and at 611 cm−1 corresponding to MXene) as a benchmark instead of additional beacon molecules. The linear goodness of fit (the determination coefficient R2) for this strategy is available in amounts up to 0.9995, which is significantly higher than that of single or double peak calibrated case, and thus improves the detection accuracy dramatically. Meanwhile, vertical MoS2 nanosheets anchored on layered MXene can provide uniformly ordered sites for accommodating suitably sized AuNPs as “hot spots” with controllable particle gap of 2.2 nm, resulting in the maximum amplified SERS signal at 1362 cm−1 generated by hairpin probe DNA with Cy5. A linear detection window from 10 am to 1 nm with an ultralow detection limit of 6.61 am for miRNA‐182 is achieved.

Surface‐enhanced Raman scattering (SERS) has received widespread attentions in biological detection since its single‐molecule sensitivity. The SERS substrate with numerous highly active hotspots can effectively capture target, whose precise fabrication is undoubtedly extremely challenging but attractive. Therefore, this work tries to achieve an ultrasensitive SERS platform based on the accurate regulations of its composition, structure, and distribution. Specifically, ordered CdS/MoS2@Au nanoparticles (AuNPs) nanopillars are fabricated by the aid of atomic layer deposition (ALD). Optimal selection and precise control of ALD cycles in MoS2 fabrication can accurately adjust the pillar diameter and obtain the most favorable minimum gap (8 nm) between pillars, which is advantageous to form multidimensional and high‐density hotspots thus generating a 9.4‐times promotion as compared with the control group. On the other hand, calculated analysis shows that appropriate spacing distance between pillars slightly larger than analyte molecule can produce considerable capillary force, which is advantageous to capture analyte and anchor it in the high‐charged‐density region for higher self‐activation of hotpots. Based on the above two favorable factors, a linear detection window from 10 × 10−18 to 1 × 10−8 m with an ultralow detection limit of 0.82 × 10−18 m for miRNA‐182 is achieved.