Comparative evaluation of freeze-dried and liquid antigens in the direct agglutination test for serodiagnosis of visceral leishmaniasis (ITMA-DAT/VL)

Abstract

Objective: The direct agglutination test (DAT) for visceral leishmaniasis (VL) with liquid (LQ) antigen is known to be only moderately reproducible because of inter-observer and batch-to-batch variability as well as its sensitivity to temperature and shaking during transport. We evaluated a DAT with freeze-dried (FD) antigen and compared it with the LQ antigen version. Methods: Blood samples of clinical VL suspects and healthy endemic controls were collected in Sudan, Nepal and India. Both test versions were performed in duplicate in the respective countries and in the reference laboratory. Interbatch variability and stability tests were conducted and agreement was examined within and between centres on a dichotomic scale by Cohen's kappa as well as on a continuous scale through Bland-Altman plots. Results: The FD antigen remains fully active even after storage at 45 °C for 24 months. Using a cut-off titre of 1 : 6400, the agreement between the FD and the LQ formats was excellent. Conclusion: The major advantages of FD antigen are its better stability at higher temperatures and its longer shelf life, which make it much more suitable than the LQ version for use in the field.