Background: While microscopy evaluation of skin scraping for detecting scabies has a 100% positive predictive value and specificity by definition, it has low sensitivity. Dermoscopy has not yet been widely accepted for diagnosis, and long-term clinician training is needed. Objectives: To evaluate the diagnostic validity of cytochrome c oxidase subunit 1 (cox1) gene nested PCR as an adjunctive method for diagnosing scabies. Methods: A total of 302 skin lesions from 50 patients suspected of scabies were examined with dermoscopic (DS) inspection and microscopic (MS) examination. Each samples was placed in a sterile microcentrifuge tube and stored at -80℃ for cox1 nested PCR analysis. Results: For all lesions, sensitivities of DS and MS were 55.9% and 55.2% respectively, using the cox1 nested PCR as 100%. The results of DS and MS were significantly different from that of cox1 nested PCR . Conclusion: The cox1 nested PCR can be utilized as an adjunctive method for diagnosing scabies, and it is more sensitive and accurate than dermoscopy or microscopy alone.