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Multiplex biosensors are highly sought-after tools in disease diagnosis. This technique involves the simultaneous sensing of multiple biomarkers, whose levels and ratios can provide a more comprehensive assessment of disease conditions compared to single biomarker detection. In most diseases like cancer due to its complexity, several biomarkers are involved in their occurrence. On the other hand, a single biomarker may be implicated in various diseases. Multiplex sensing employs various techniques, such as optical, electrochemical, and electrochemiluminescence methods. This comprehensive review focuses on optical multiplex sensing techniques, including surface plasmon resonance, localized surface plasmon resonance, fluorescence resonance energy transfer, chemiluminescence, surface-enhanced Raman spectroscopy, and photonic crystal sensors. The review delves into their mechanisms, materials utilized, and strategies for biomarker detection., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Parvin Samadi Pakchin reports financial support was provided by Iran National Science Foundation INSF grant number 99029087. If there are other authors, they 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 © 2024 Elsevier B.V. All rights reserved.)