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RhoGam, also known as Rh immunoglobulin, is the primary intervention for unsensitized Rh-negative mothers during the third trimester to prevent Rh sensitization. If an Rh-negative mother is exposed to Rh-positive fetal blood cells, she could develop antibodies against the Rh factor, leading to complications in current or future pregnancies. By administering RhoGam during the third trimester, healthcare providers effectively inhibit the mother's immune response to Rh-positive red blood cells, thereby reducing the risk of hemolytic disease of the newborn in subsequent pregnancies.

This intervention is specifically important to carry out around the 28th week of gestation. Other options listed, such as ultrasound and amniocentesis, are valuable diagnostic tools but do not address the prevention of Rh sensitization. While the glucose tolerance test is essential for assessing gestational diabetes, it does not pertain to the risk of Rh incompatibility. Therefore, RhoGam plays a crucial, preventive role for unsensitized Rh-negative mothers in the third trimester.