The development of convenient and sensitive multi-readout immunoassay is crucial but highly challenged for meeting the demand of exactness and diversity in early clinical diagnosis. Herein, a split-type multiple stimuli-responsive biosensor was outlined combined with the outstanding superiority of luminol probe-based electrochemiluminescence (ECL) strategy, mimicking enzyme-mediated colorimetric system and portable photothermal effect-induced temperature sensing. Especially, versatile MoS nanosheets (MoS NSs) with distinguished property not only acted as dual-promoter to improve the cathodic ECL of luminol because of its good electrocatalytic activity for dissolved O and favorable photothermal effect for elevating electrode temperature, but also used as nanozyme to regulate subsequent split-type visual colorimetric sensing due to its peroxidase-like activity for the generation of oxidized 2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) in ABTS-HO colorimetric system. More importantly, the green oxidized ABTS (ABTS) also exhibited strong near-infrared (NIR) laser-triggered photothermal performance, which can be innovatively employed as sensitive photothermal agent for converting biological signals into temperature under the irradiation of NIR laser, accomplishing more simpler temperature quantitative detection by a portable thermometer. Furthermore, on account of the affinity discrepancy of MoS NSs to single-stranded and double-stranded nucleic acids, a label-free proximity hybridization-based multifunctional assay platform was proposed for target detection with human epididymis-specific protein 4 (HE4) as model protein, demonstrating good analytical performances. Significantly, this innovative work not only enriches the foundational study of multi-model biosensing based on the unitary material but also provides an unambiguous guideline for exploring more accurate and simpler point-of-care diagnosis.