A novel method of species identification using human muscle-specific beta-enolase

A novel method of species identification using a sensitive sandwich enzyme immunoassay for human muscle-specific beta-enolase was used to analyze bloodstains and skeletal muscle. Human beta-enolase is detectable in human blood at dilutions up to 3 x 10(3) and human muscle extract (0.1 g/ml) at dilutions up to 5 x 10(6) using a sandwich enzyme immunoassay. The cross-reactivities of both blood and skeletal muscle extracts from other species were 0.01 or less. When human skeletal muscle extract was added in a final concentration of 10 mg/ml, the ratio of beta-enolase to total protein in bloodstains was approximately 1000, whereas the addition of skeletal muscle extract of other species resulted in a ratio of less than 10. Application of this method in a practical case proved that human muscle-specific beta-enolase as a marker for species identification is very effective in forensic practice.