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This paper describes a single tube reverse transcription-polymerase chain reaction (RT-PCR) method for detection of African horse sickness virus (AHSV). The genomic segments 10 of viruses of the 9 AHSV serotypes were amplified. The 758bp products were digested to completion by restriction enzymes. The restriction fragment length polymorphisms of segments 7 and 10 cDNA allowed the differentiation of the nine serotypes.