USE OF A SEQUENCE CHARACTERIZED AMPLIFIED REGIONS (SCARS) - MULTIPLEX PCR METHOD TO IDENTIFY EIMERIA SPECIES OF CHICKENS FROM WIDELY DISTRIBUTED GEOGRAPHIC AREAS

摘要

A Random Amplified Polymorphic DNA-Sequence Characterized Amplified Regions (RAPD-SCAR) maker based Multiplex-Polymerase Chain Reaction (PCR) was evaluated as a potential method of differentiating species of Eimeria oocysts in chickens. Oocysts of chicken coccidia were obtained from different geographical areas of 3 continents (Africa, North America and Europe) and genomic DNA was extracted. PCR was performed using pairs of RAPD-SCAR based primers that specifically amplify genomic DNA from six species of Eimeria found in chickens (E. maxima, E. tenella, E. acervulina, E. mitis, E. necatrix and E. brunetti) in a single reaction tube. Results showed that the method is useful in differentiating species of coccidia obtained from Nigeria, the United Kingdom, United States of America and Canada. Samples containing more than one species of Eimeria were successfully detected by the method despite low numbers of oocysts in some of the samples (less than 1000 oocysts per ml of isolate). The method demonstrates the simultaneous detection of the 6 Eimeria species affecting chickens in a single-tube multiplex PCR either as single or mixed infections; and offers a universal tool for the epidemiological study of coccidiosis in poultry.