EIAV Effectiveness Detection by Nested PCR Comparing two Different Samples: PBMCs and Bronchoalveolar Wash

Abstract

Problem statement: Equine Infectious Anemia (EIA) is a chronic, relapsing infectious disease of horses caused by Equine Infectious Anemia Virus (EIAV). The objective of this experiment was compared EIAV effectiveness detection in Peripheral Blood Mononuclear Cells (PBMCs) and Bronchoalveolar wash (BAW) from naturally infected horses. Approach: Fifty seven DNA samples isolated from PBMCs and BAW of naturally infected animals were used for a nested PCR amplifying a 408 bp gag gene fragment region. Equine Dermal cells (ED) infected and non infected by EIAV was used as nested PCR negative and positive control. Horses’ blood samples were firstly tested as positive in Agar Gel Immunodiffusion Assay (AGID). Results: Results demonstrated 89% (51/57) of gag gene amplification in PBMCs samples and only 47% (27/57) of gag gene amplification in BAW samples. Conclusion: The nested PCR assay used in the present study detected more EIAV positive samples in PBMC than in BAW, indicating PBMCs as a reliable source for EIAV diagnosis.