Mamat U, Löbau S, Persson K, Brade H
Institut für Experimentelle Biologie und Medizin, Forschungsinstitut Borstel, Germany.
Microb Pathog. 1994 Aug;17(2):87-97. doi: 10.1006/mpat.1994.1055.
The gene gseA, involved in the expression of the genus-specific epitope of chlamydial lipopolysaccharide (LPS), was analyzed by temperature gradient gel electrophoresis (TGGE) to visualize nucleotide sequence variations among the 15 serovars of Chlamydia trachomatis. Sequence analysis showed that the TGGE melting-profile patterns were able to detect single nucleotide variations within gseA and allowed the arrangement of the serovars in groups of both identical nucleotide sequences and sequences containing identical sites of nucleotide substitutions. Compared to serovar L2, four types of patterns were obtained: (i) serotypes A and Ba; (ii) B and C (causing endemic trachoma); (iii) D through K (causing sexually transmitted oculo-genital infections); (iv) L1 through L3 (the causative agents of lymphogranuloma venereum). A total of 58 isolated of C. trachomatis of genital or conjunctival origin were tested by this method in comparison to reference strains. Forty-eight isolates (13 of type E, 16 of type F, nine of type G, and ten of type K) yielded the same melting profile as the corresponding type strain, independent of whether they were isolated from genital or ocular infections. However, ten B serotype strains of genital origin behaved in TGGE like a typical genital strain and not a trachoma strain. Thus, although gseA was found to be highly conserved among C. trachomatis, the obtained TGGE profiles of the tested strains tended to correlate with their specific site of infection.
通过温度梯度凝胶电泳(TGGE)分析参与沙眼衣原体脂多糖(LPS)属特异性表位表达的基因gseA,以观察沙眼衣原体15个血清型之间的核苷酸序列变异。序列分析表明,TGGE解链图谱模式能够检测gseA内的单核苷酸变异,并能将血清型按照相同核苷酸序列组以及含有相同核苷酸取代位点的序列组进行排列。与血清型L2相比,获得了四种模式:(i)血清型A和Ba;(ii)B和C(引起地方性沙眼);(iii)D至K(引起性传播的眼-生殖器感染);(iv)L1至L3(性病性淋巴肉芽肿的病原体)。与参考菌株相比,用该方法共检测了58株来自生殖器或结膜来源的沙眼衣原体。48株分离株(13株E型、16株F型、9株G型和10株K型)产生了与相应标准菌株相同的解链图谱,无论它们是从生殖器感染还是眼部感染中分离得到的。然而,10株来自生殖器的B血清型菌株在TGGE中的表现类似于典型的生殖器菌株,而不是沙眼菌株。因此,尽管发现gseA在沙眼衣原体中高度保守,但所获得的受试菌株的TGGE图谱倾向于与其特定感染部位相关。