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• The study establishes a novel assay for detection of canine astrovirus. • The method in this study is highly sensitive, specific, and reliable. • The method is of great significance for the clinical detection of canine astrovirus. In this study, a TaqMan-based real-time polymerase chain reaction (PCR) method to detect canine astrovirus in clinical samples was developed. Primers and probes were designed to target conserved regions of the complete viral genome sequence. The results showed that the proposed method can detect a minimum of 101 copy numbers. No cross-reactivity with other canine and feline viruses was observed. The coefficient of variation was <5%. Evaluation of the clinical samples showed that quantitative PCR had a 5.26 % higher positive detection rate than conventional PCR. These results indicate that the method developed in this study is highly reliable and suitable for veterinary clinical diagnosis and epidemiological investigations. [ABSTRACT FROM AUTHOR]