Ariyoshi Rikako, Kawai Toru, Honda Takashi, Tokiyoshi Sachio
Animal Vaccine Production Department, The Chemo-Sero-Therapeutic Research Institute, Okubo, Kumamoto.
J Vet Med Sci. 2010 Jun;72(6):687-92. doi: 10.1292/jvms.09-0130. Epub 2010 Jan 22.
In this study, we attempted to establish a simple detection method for classification of IBV S1 genotypes by direct reverse transcriptase-polymerase chain reaction (RT-PCR). Then, to evaluate the usefulness of the S1 genotype-specific RT-PCR, we examined the relationship between S1 genotypes and serotypes of IBV in Japan. Sequencing of the S1 genes of IBV and phylogenetic tree analysis were conducted. On the basis of the sequencing data of the S1 genotype samples, we determined primer sets specific for each genotype. Five vaccine strains in Japan as reference strains and 46 field isolates were classified into different genetic clusters by phylogenetic tree analysis (JP-1, JP-II, JP-III, Mass and 4/91) and were matched to the results of S1 genotype-specific RT-PCR. A cross virus-neutralizing test showed that the five vaccine strains in Japan exhibited different serotypes from each other. The concordance rate of the 46 field isolates between the S1 genotypes and serotypes was 65.2%. The present study indicates that genotype-specific RT-PCR could be a convenient and useful tool for determining IBV serotypes and could contribute to the control of IBV outbreaks in Japan.
在本研究中，我们试图通过直接逆转录聚合酶链反应（RT-PCR）建立一种用于IBV S1基因型分类的简单检测方法。然后，为了评估S1基因型特异性RT-PCR的实用性，我们研究了日本IBV的S1基因型与血清型之间的关系。进行了IBV S1基因的测序和系统发育树分析。基于S1基因型样本的测序数据，我们确定了针对每种基因型的引物组。通过系统发育树分析（JP-1、JP-II、JP-III、Mass和4/91），将日本的5种疫苗株作为参考株和46株田间分离株分为不同的遗传簇，并与S1基因型特异性RT-PCR的结果相匹配。交叉病毒中和试验表明，日本的5种疫苗株彼此表现出不同的血清型。46株田间分离株的S1基因型与血清型之间的符合率为65.2%。本研究表明，基因型特异性RT-PCR可能是确定IBV血清型的一种方便且有用的工具，并有助于控制日本的IBV疫情。
