Dezhbord Mehrangiz, Esmaelizad Majid, Khaki Pejvak, Fotohi Fariba, Zarehparvar Moghaddam Athena
Razi Vaccine and Serum Research Institute, Karaj, Iran.
J Infect Dev Ctries. 2014 Jun 11;8(6):688-93. doi: 10.3855/jidc.3174.
Leptospirosis, caused by infection with pathogenic Leptospira species, is one of the most prevalent zoonotic diseases in the world. Current leptospiral vaccines are mainly multivalent dead whole-cell mixtures made of several local dominant serovars. Therefore, design and construction of an efficient recombinant vaccine for leptospirosis control is very important. OmpL1 is an immunogenic porin protein that could be of special significance in vaccination and serodiagnosis for leptospirosis.
Three strains belonging to pathogenic L. interrogans were analyzed. The specific primers for proliferation of the ompL1 gene were designed. The amplified gene was cloned. In order to investigate the ompL1 nucleotide sequence and homological analysis of this gene, ompL1 genes cloned from standard vaccinal Leptospira serovars prevalent in Iran were sequenced and cloned.
PCR amplification of the ompL1 gene using the designed primers resulted in a 963 bp ompL1 gene product. The PCR based on the ompL1 gene detected all pathogenic reference serovars of Leptospira spp. tested. Based on alignment and phylogenetic analysis, although the ompL1 nucleotide sequence was slightly different within three vaccinal serovars (100%-85% identity), amino acid alignment of the OmpL1 proteins revealed that there would be inconsiderable difference among them.
The ompL1 gene of the three isolates was well conserved, differing only by a total of 6 bp and the proteins by 2 amino acids. The cloned gene could be further used for expression and recombinant OmpL1 as an efficient and conserved antigen, and may be a useful vaccine candidate against leptospirosis in our region.
钩端螺旋体病由致病性钩端螺旋体属感染引起,是世界上最普遍的人畜共患病之一。目前的钩端螺旋体疫苗主要是由几种当地优势血清型制成的多价死全细胞混合物。因此,设计和构建一种有效的重组疫苗来控制钩端螺旋体病非常重要。外膜脂蛋白L1(OmpL1)是一种具有免疫原性的孔蛋白,在钩端螺旋体病的疫苗接种和血清学诊断中可能具有特殊意义。
对三株问号钩端螺旋体进行分析。设计用于扩增ompL1基因的特异性引物。对扩增的基因进行克隆。为了研究ompL1核苷酸序列及其同源性分析,对从伊朗流行的标准疫苗钩端螺旋体血清型中克隆的ompL1基因进行测序和克隆。
使用设计的引物对ompL1基因进行PCR扩增,得到一个963bp的ompL1基因产物。基于ompL1基因的PCR检测了所有测试的致病性钩端螺旋体参考血清型。基于比对和系统发育分析,虽然三个疫苗血清型内的ompL1核苷酸序列略有不同(同一性为100%-85%),但OmpL1蛋白的氨基酸比对显示它们之间的差异不大。
三株分离株的ompL1基因高度保守,总共仅相差6个碱基对,蛋白质相差2个氨基酸。克隆的基因可进一步用于表达,重组OmpL1作为一种高效且保守的抗原,可能是我们地区抗钩端螺旋体病的有用候选疫苗。