Development of a polymerase chain reaction assay to identify Eimeria maxima oocysts.

摘要

Coccidiosis, is an endemic poultry disease, which easily spreads from farm to farm. Three detection methods exist to determine Eimeria species. Problems arise when the methods produce inconclusive results. A new method, the random-amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR)-based assay, was used to initiate screening for polymorphic deoxiribonucleic acids of Eimeria maxima. The genomic DNA was obtained from poultry propagated, recovered, and sporulated oocysts. The oocysts were crushed with glass beads and subsequently purified by phenol/chloroform extraction and ethanol precipitation. PCR amplifications using 28 arbitrary oligonucleotides were performed. Characteristic banding patterns of Eimeria maxima were generated through agarose gel electrophoresis. The Eimeria maxima DNA bands were then compared to banding patterns produced by Eimeria tenella DNA. The result was 23 unique bands generated from 10 of the 31 primers. This work demonstrates the potential of PCR-based assays for identification and detection of Eimeria maxima.