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Context: It has been hypothesized that diagonal earlobe crease (DELC), “Frank's sign” is indicative of coronary artery disease (CAD) and/or diabetes mellitus (DM). Several studies have confirmed an association between DELC and cardiac morbidity, mortality, and hypertension (HTN). However, some studies have not found any significant association. Aims: This study aims to find out the prevalence of DELC and its association with CAD, DM, and HTN. Settings and Design: Sangli-Miraj-Kupwad Corporation area. This was a cross-sectional analytical study. Subjects and Methods: Study participants: Adults from 18 to 60 years age. Inclusion criteria: willing to participate in the study Exclusion criteria: Wearing heavy ear rings and excessive normal generalized wrinkling of the skin. Sample size: Sample size 6310, determined after a pilot study revealing DELC in 1.5%. Sampling technique: Two-stage cluster sampling. Duration of study: 6 months. Study tools: Predesigned, pilot tested pro forma. Statistical Analysis: Statistical analysis was done by using SPSS 22 software. Prevalence and percentages were calculated, and Chi-square test was applied. Results: Out of 6638 participants, 179 had DELC. The prevalence of bilateral DELC was 2.7%. The prevalence was significantly high among males (4.13%) and in the 51–60 years age group (5.29%). The prevalence of Grade 3 DELC was high and 91% of young adults had Grade 3 DELC. There were 408 (6.15%) participants who gave a history of CAD, 827 (12.46%) of DM, and 670 (10.09%) HTN. Significantly high association observed between DELC and CAD, DM, and HTN. CAD, DM, and HTN were significantly associated with Grade 3. Conclusions: The prevalence of bilateral DELC was 2.7% and is significantly associated with CAD, DM, and HTN.