Tham K M, Barnes S M, Hunter S N
Virology Section, Central Animal Health Laboratory, MAF Quality Management, Upper Hutt, New Zealand.
Virus Genes. 1999;18(3):235-42. doi: 10.1023/a:1008020303036.
A reverse transcription-polymerase chain reaction (RT-PCR) amplification assay was used to detect calicivirus gene sequences in a liver tissue derived from a feral rabbit which died of a recent outbreak of rabbit haemorrhagic disease (RHD) in New Zealand. Five pairs of primers were designed to amplify five complementary DNA genomic sequence stretching from nucleotide positions 1594 to 7071, yielding amplified fragments of 361, 340, 805,670 and 386 bp for the primer pairs RC-1/RC-2, RC-3/RC-4, RC-5/RC-6, RC-7/RC-8 and RC-9/RC-10 respectively. The identity of the amplified fragments was confirmed by chemiluminescence Southern blot hybridization and direct cycle sequencing. The nucleotide sequences of the five amplified fragments were determined and comparisons of the nucleotide and deduced amino acid sequences revealed a close genetic relationship of the New Zealand isolate 97-10372 with overseas strains of RHD virus.
采用逆转录聚合酶链反应（RT-PCR）扩增试验，检测来自一只死于新西兰近期兔出血性疾病（RHD）暴发的野生兔肝脏组织中的杯状病毒基因序列。设计了五对引物，用于扩增从核苷酸位置1594至7071延伸的五个互补DNA基因组序列，引物对RC-1/RC-2、RC-3/RC-4、RC-5/RC-6、RC-7/RC-8和RC-9/RC-10分别产生361、340、805、670和386 bp的扩增片段。通过化学发光Southern印迹杂交和直接循环测序确认扩增片段的同一性。测定了五个扩增片段的核苷酸序列，核苷酸和推导氨基酸序列的比较显示新西兰分离株97-10372与海外RHD病毒株具有密切的遗传关系。
