Duplex rRT-PCR was established for two goose astrovirus (GAstV) genotypes.

The assay showed high specificity and sensitivity, and good reproducibility.

The assay was validated in experimental infections and clinical samples.

This rRT-PCR method will improve the molecular epidemiology of GAstV.

Goose astrovirus (GAstV) is a highly infectious pathogen that causes gout in goslings (<15 old) with typical symptoms of white urate disposition on the surface of the visceral organs and articular cavity, and a high mortality rate up to 50 %. To establish a real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay for the rapid detection of the two GastV genotypes(GAstV-1 and GAstV-2), two pairs of primers and a pair of matching TaqMan probes were designed based on conserved regions of the ORF1b gene. The established duplex rRT-PCR assay showed no cross-reactivity with 10 other common waterfowl pathogens. The minimum detection limit was 10 copies/reaction for both GAstV-1 and GAstV-2. To validate the assay, 36 cloacal swabs from experimentally infected goslings and 33 field clinical samples were tested. The assay results of the experimentally infected goslings matched the infection scheme. The positive rates of GAstV-1 and GAstV-2 in the field clinical samples were 36.36 % and 54.55 %, respectively, and the co-infection rate of the two viruses was 21.21 % based on the duplex rRT-PCR assay. In conclusion, the established assay represents a specific, sensitive, and convenient tool for detecting GAstV-1, GAstV-2, and their co-infections, and for conducting epidemiological surveys.

goose-origin H9N2 subtype avian influenza virus

conventional reverse transcription-polymerase chain reaction

coefficient of variation

muscovy duck parvovirus

goose-origin Newcastle disease virus

Riemerella anatipestifer

real-time reverse transcription-polymerase chain reaction

Goose astrovirus-1

Goose astrovirus-2

Duplex real-time quantitative reverse transcription-PCR

TaqMan probe

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