Neonatal alloimmune thrombocytopenia (NAIT) is an important hematological disorder in neonates. The pregnant mother's immune system gets sensitized to antigens expressed on fetal platelets that have been inherited from the father and begins producing specific alloantibodies against these antigens. Some of these antibodies get transported across the placenta into the baby and can damage/destroy platelets to cause fetal/neonatal thrombocytopenia. Many of these fetuses/infants develop major clinical complications such as intracranial hemorrhages. In this article, we describe normal platelet counts in neonates, the pathogenesis and epidemiology of NAIT, specific platelet antigens that have been identified as targets in NAIT, and the approach for laboratory diagnosis of NAIT. From the perspective of a transfusion medicine service, there are two targets as follows: (a) To identify the differences in the antigenic profiles of the platelets of the mother and her fetus/infant and (b) To detect alloantibodies in the maternal serum that may be specifically reactive to these platelet antigens. Early identification of NAIT can help timely institution of appropriate treatment. In this project, we reviewed the laboratory profiles of infants who were diagnosed to have NAIT at our own institution and also mined the literature in the databases EMBASE, PubMed, and Scopus.