Greiser-Wilke I, Depner K, Fritzemeier J, Haas L, Moennig V
Institute of Virology, Veterinary School Hannover, Germany.
J Virol Methods. 1998 Nov;75(2):141-50. doi: 10.1016/s0166-0934(98)00109-8.
The commercial software program HLA SequiTyper (Amersham Pharmacia Biotech), designed originally for human leukocyte antigen typing, was adapted for rapid typing of classical swine fever (CSF) virus isolates. The program compares new sequence data with those stored in a database file and calculates the most probable assignment. For generating the CSF virus sequence database, 150 bp of the 5' nontranslated genomic region (5'-NTR) from 88 German classical swine fever virus isolates from outbreaks between 1984 and 1997 were solid-phase sequenced directly after RT-PCR amplification. Sequence alignments showed that they all belonged to the previously defined genetic group 2. Within this group, six different subgroups could be distinguished, and were designated according to the geographic location where they are either still endemic or where they appeared most commonly. The advantage of using the HLA SequiTyper program is that it reads directly the sequence files as generated by the ALF sequencer (Amersham Pharmacia Biotech), making any manipulations unnecessary. In addition, a constant quality control of the raw sequence data can be achieved, as more than one sequence from the same isolate can be evaluated at once. Using this approach, new CSF isolates can be typed within 2 days.
最初设计用于人类白细胞抗原分型的商业软件程序HLA SequiTyper(安玛西亚生物科技公司),被改编用于经典猪瘟(CSF)病毒分离株的快速分型。该程序将新的序列数据与存储在数据库文件中的数据进行比较,并计算最可能的分型。为了生成CSF病毒序列数据库,对1984年至1997年间德国88株来自疫情爆发的经典猪瘟病毒分离株的5'非翻译基因组区域(5'-NTR)的150 bp进行了RT-PCR扩增后直接进行固相测序。序列比对表明它们都属于先前定义的遗传组2。在这个组内,可以区分出六个不同的亚组,并根据它们仍然流行或最常出现的地理位置进行命名。使用HLA SequiTyper程序的优点是它可以直接读取由ALF测序仪(安玛西亚生物科技公司)生成的序列文件,无需进行任何操作。此外,由于可以一次评估来自同一分离株的多个序列,因此可以对原始序列数据进行持续的质量控制。使用这种方法,新的CSF分离株可以在2天内完成分型。