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Aflatoxin B1 (AFB1) as the most toxic mycotoxin in contaminated food can greatly threaten human health, and sensitive and selective detection of AFB1 is thus highly desired. An ultrasensitive surface-enhanced Raman spectroscopy (SERS) aptasensor was developed for AFB1 detection in peanut oil samples. SH-cDNA modified Fe 3 O 4 @Au nanoflowers acted as capture probes, SH-Apt modified Au@Ag nanospheres and commercial Cy3-Apt were used as reporter probes. Strong SERS signals of reporter probes were produced due to the recognition of AFB1 aptamer and its complementary strand (SH-cDNA). With the preferred binding of AFB1 aptamer to AFB1, reporter probes were released from capture probes, causing a linear decrease in SERS intensity. Therefore an ultralow detection limit of 0.40 pg·mL -1 in a wide linear range of 0.0001-100 ng·mL -1 was obtained and the sensibility of this SERS aptasensor was higher than that of the Cy3-Apt based SERS aptasensor. In addition, an excellent selectivity in interfering toxins and satisfactory recoveries of 96.6-115% in peanut oil samples were obtained, proving this aptasensor is a promising analytical tool in AFB1 detection., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)