Genetic characterization of GRA6 genes from Toxoplasma gondii from pigs in Okinawa, Japan
Genetic characterization of GRA6 genes from Toxoplasma gondii from pigs in Okinawa, Japan
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DOI：
10.1292/jvms.68.1105
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发表时间：
2006-10-01
影响因子：
1.2
通讯作者：
Fujisaki, Kozo
中科院分区：
文献类型：
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作者：
Zakimi, Satoshi;Kyan, Hisako;Fujisaki, Kozo
采用巢式PCR-限制性片段长度多态性（RFLP）和GRA 6基因序列分析方法对冲绳县猪弓形虫病进行了研究。采用巢式PCR方法，从屠宰场采集的，头猪的91份淋巴结样本中，有33份检出了弓形虫DNA，其病变与弓形虫病样本中发现的病变相似。对33份PCR阳性标本中的29份进行了MseI限制性片段长度多态性（RFLP）分析，将分离株分为T.刚地。29例研究样本的基因分型结果如下：1型13例（44.8%），11型14例（48.3%），111型2例（6.9%）。对12株冲绳分离株的GRA 6基因进行了克隆和序列测定。发现了9个新的核苷酸序列，并在13个位置发现了冲绳分离株特异性的核苷酸替换。系统发育分析表明，GRA 6基因可分为3个主要类群，其中冲绳分离株的GRA 6基因型II和III分别与Beverley株和NED株有较近的亲缘关系。本研究结果为T.日本国内的弓形虫感染。
Toxoplasma gondii from pigs in Okinawa Prefecture was characterized by nested PCR-restriction fragment length polymorphism (RFLP) and DNA sequence analysis of the dense granule antigen GRA6 gene. By nested PCR, parasite DNA was detected in 33 out of 91 lymph node samples with lesions similar to those found in toxoplasmosis samples that had been collected from,pigs at an abattoir. RFLP analysis with MseI was successfully conducted in 29 of 33 PCR-positive samples to group the isolates into one of the three genotypes of T. gondii. Genotyping of the 29 studied samples rendered the following results: 13 of type 1 (44.8%), 14 of type 11 (48.3%), and 2 of type 111 (6.9%). The GRA6 genes of 12 Okinawa isolates were cloned and sequenced. Nine new nucleotide sequences were found, and nucleotide substitutions specific for the Okinawa isolates were found at 13 positions. Phylogenetic analysis indicated that all GRA6 sequences were divided into one of the 3 main groups, and Okinawa isolates of GRA6 genotypes II and III seemed to be closely related to the Beverley strain and the NED strain, respectively. The results from this study may provide basic and useful information for the analysis of the molecular epidemiology of T. gondii infection within Japan.