The Human Leukocyte Antigen (HLA) system forms the central part of the immune system and is crucial in the recognition and elimination of “non-self” antigens. While this role of the HLA system is essential in the effective defense of the organism against pathogens, it is undesirable in organ and tissue transplantation because it enables the recognition of mismatched HLA molecules of the donor as being foreign and stimulates the graft rejection reaction. Organ transplantation involves the introduction of antigens that are more or less mismatched to the recipient; therefore, in order to achieve the best possible match in the HLA system between the recipient and the donor, a whole series of immunogenetic tests is performed, including crossmatching (XM). If performed before kidney transplantation, it represents the final in vitro test to rule out the presence of donor-specific antibodies, which may cause graft rejection and which may not have been detected by earlier serum screening. The beginning of XM was marked by the complement-dependent cytotoxicity (CDC) method developed by Terasaki and colleagues in 1964. Later, as a result of advances in technology and the need for methods that overcome the limitations of CDC, flow cytometry and Luminex XM assays were developed. The introduction of solid-phase technology brought a new dimension to the detection of low-level HLA antibodies and the determination of their specificities, which enabled the development and implementation of the virtual XM test (vXM). It is an in silico test that assesses the immunological match between the recipient and the organ donor based on the analysis of the specificity of the antibodies present in the recipient’s serum and the HLA typing of the organ donor. Each method has its own advantages and limitations, which are described below and need to be taken into account, considering their significant impact on clinical application in kidney transplantation.