METHOD FOR DIFFERENTIALLY DIAGNOSING DUCK HEPATITIS VIRUS TYPE 1

摘要

A differential diagnostic method of duck hepatitis virus(DHV) type 1 is provided to improve rapidness and accuracy of diagnosis, and diagnose common genes and different genes of duck hepatitis virus type 1 in the liver tissue separated from young ducks infected by the duck hepatitis virus type 1, and viruses causing other RNA viral diseases derived from fowls. Duck hepatitis virus type 1 gene has the nucleotide sequence consisting of 2C gene, capsid gene and 5'-UTR(untranslated region) gene, wherein the 2C gene has the nucleotide sequence selected from SEQ ID NOs:1, 4, 7, 10, 13, 16, 19, 22 and 25; the capsid gene has the nucleotide sequence selected from SEQ ID NOs:2, 5, 8, 11, 14, 17, 20, 23 and 26; and the 5'-UTR gene has the nucleotide sequence selected from SEQ ID NOs:3, 6, 9, 12, 15, 18, 21, 24 and 27. Duck hepatitis virus-specific primers have the nucleotide sequences of SEQ ID NOs:28 to 33 and specifically recognize the 2C gene, capsid gene and 5'-UTR gene, respectively. A differential diagnostic method of duck hepatitis virus(DHV) type 1 comprises the steps of: (1) homogenizing the liver tissue separated from young ducks infected by the duck hepatitis virus type 1 to float in 10% PBS(phosphate buffered saline); (2) obtaining the supernatant and separating RNA from the supernatant by RNA extraction method; (3) synthesizing cDNA(complementary DNA) from the separated RNA through reverse transcription method by using reverse transcriptase; (4) preparing primers specific to 2C gene, capsid gene and 5'-UTR gene from cDNA, and amplifying the genes through PCR(polymerase chain reaction); and (5) subjecting the PCR products to electrophoresis using 1.5% agarose gel.