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Simple Summary: Children who inherit a TP53 (tumor suppressor) mutation tend to develop adrenocortical tumor (ACT) in the initial years of life, requiring genetic counseling and DNA testing after birth. In 2001, a TP53 mutation (R337H) was identified in Paraná State (South Brazil) and was later confirmed to be highly frequent in South Brazil, where the pediatric ACT incidence is up to 20 times higher than that in other countries. We evaluated anxiety and depression in mothers of these newborns to determine the effect of genetic testing. We found that anxiety (but not depression) is presented by mothers of R337H-positive newborns for less than 4 months. These findings were more common in mothers with previous mental health vulnerabilities. The anxiety test scores must be differentiated, and appropriate psychological support should be provided for these mothers and for mothers of newborns inheriting other types of TP53 mutation in other countries. Counselling and genetic testing (CGT) after neonatal screening may increase depression and anxiety (DA) levels during cancer surveillance. This study assessed the DA scores in mothers of newborns from Paraná state, Southern Brazil, carrying the TP53 p.R337H variant. To understand and adjust DA conditions during term of pregnancy, we initially detected sociodemographic covariates [marital status (MS), number of children (NC), and/or education level (EL): MS-NC-EL] on an independent group of pregnant women (not subjected to genetic testing). The Hospital Anxiety and Depression Scale (HADS) was used to assess risk factors in pregnant (cross-sectional analysis) and unrelated mothers (at 2-month intervals, longitudinal study) of TP53 p.R337H-tested newborns (three sessions of HADS analysis) using Wilcoxon (Mann–Whitney) and Kruskal–Wallis nonparametric tests. Lower anxiety levels were observed in mothers of noncarriers (without MS-NC-EL = 6.91 ± 1.19; with MS-NC-EL = 6.82 ± 0.93) than in mothers of p.R337H carriers in the first session (without MS-NC-EL = 6.82 = 8.49 ± 0.6025, with MS-NC-EL = 6.82 = 9.21 ± 0.66). The anxiety levels significantly decreased 4 months after CGT (third session) in mothers of p.R337H carriers. We did not find a significant change in depression scores. Mothers with mental health instability requiring medications need periodical psychological support during and after CGT. [ABSTRACT FROM AUTHOR]