Etemady Afshar, Mohammdi Mohsen, Esmaelizad Majid, Alamian Saeed, Vahedi Fatemeh, Aghaeipour Khosro, Behrozikhah Ali Mohammad, Faghihloo Ebrahim, Afshar Davoud, Firuzyar Sajad, Rahimi Arian
Ph.D. of Medical Bacteriology, Department of Brucellosis, Razi Vaccine and Serum Research Institute, Karaj, Iran.
Assistant Professor, Department of Pharmaceutical Biotechnology, Faculty of pharmacy, Lorestan University of Medical sciences, Khoramabad, Iran.
Electron Physician. 2015 Oct 19;7(6):1381-6. doi: 10.14661/1381. eCollection 2015 Oct.
Brucella spp. are gram-negative, facultative intracellular bacteria pathogens responsible for brucellosis, a zoonotic disease that can cause abortion, fetal death, and genital infections in animals and undulant fever in humans. Lipopolysaccharide (LPS) is known as a major virulence factor of Brucella spp. The wboA gene is capable of encoding a glycosyltransferase that appears to play a major role in LPS biosynthesis. Hence, the characterization of this gene can help in the clarification of the pathogenicity of Brucella spp.
This study was carried out at Razi Vaccine and Serum Research Institute in 2011. Briefly, the wboA gene in B. abortus biovar 3 and B. melitensis biovar 1, the predominant biovars in Iran, were amplified by using two pairs of specific primers. Polymerase chain reaction (PCR) products were cloned into a thymine-adenine (TA) cloning vector and transformed into an E. coli DH5α before being sequenced. Multiple alignments of identified sequences were performed, with all wboA sequences deposited in the GenBank sequence database.
This study showed that a mismatch has occurred in B. melitensis biovar 1; this biovar is predominant in Iran. In contrast, the wboA gene from B. abortus biovar 3 was similar to that of other B. abortus variations.
The comparison and alignment of the wboA gene of native Brucella strains in Iran to all wboA sequences deposited in GenBank revealed that the wboA gene has changed in the long term; hence, because of its unique nucleotide pattern, the gene can be used for specific diagnosis of B. abortus and B. canis.
布鲁氏菌属是革兰氏阴性兼性细胞内细菌病原体,可导致布鲁氏菌病,这是一种人畜共患病,可引起动物流产、胎儿死亡和生殖系统感染以及人类波浪热。脂多糖(LPS)是布鲁氏菌属的主要毒力因子。wboA基因能够编码一种糖基转移酶,该酶似乎在LPS生物合成中起主要作用。因此,对该基因的表征有助于阐明布鲁氏菌属的致病性。
本研究于2011年在拉齐疫苗和血清研究所开展。简要来说,使用两对特异性引物扩增伊朗主要生物变种流产布鲁氏菌生物变种3和马尔他布鲁氏菌生物变种1中的wboA基因。聚合酶链反应(PCR)产物被克隆到胸腺嘧啶-腺嘌呤(TA)克隆载体中,并在测序前转化到大肠杆菌DH5α中。对鉴定出的序列进行多重比对,并将所有wboA序列存入GenBank序列数据库。
本研究表明,在伊朗占主导地位的马尔他布鲁氏菌生物变种1中发生了错配。相比之下,流产布鲁氏菌生物变种3的wboA基因与其他流产布鲁氏菌变种的相似。
将伊朗本地布鲁氏菌菌株的wboA基因与GenBank中存入的所有wboA序列进行比较和比对后发现,wboA基因长期以来发生了变化;因此,由于其独特的核苷酸模式,该基因可用于流产布鲁氏菌和犬布鲁氏菌的特异性诊断。