Ji W S, Hu J L, Qiu J W, Pan B R, Peng D R, Shi B L, Zhou S J, Wu K C, Fan D M
Chinese PLA Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
World J Gastroenterol. 2001 Dec;7(6):864-7. doi: 10.3748/wjg.v7.i6.864.
To discover the relationship between the genotype and antigen serotype of flagellin C among Salmonella strains.
Fragment of Salmonella flagellin C in plasmid pLS408 was cloned, sequenced and compared with the corresponding sequence in other strains. Salmonella strains including two typhi strains, one paratyphoid strain, one enteritidis and one typhimurium strain were isolated from outpatients. Genome DNA was purified respectively from these clinical isolates, then the corresponding flagellin C fragment was amplified by polymerase chain reaction,and the amplification products were analyzed by agarose gel electrophoresis.
The cloned fragment includes 582 nucleotides encoding the variable region and partial conservative region of Salmonella flagellin C in plasmid pLS408. With comparison to the corresponding sequences reported previously, there is only a little difference from other strains with the same flagellar serotype in both nucleotide and amino acid level. Specific PCR products were amplified in Salmonella strains with flagellar serotype H-1-d including S. muenchen, typhi and typhimurium, but not in S. paratyphoid C or S. enteritidis strains.
In this experiment, the specificity of nucleotide sequence could be found in flagellin C central variable regions as it exists in flagellar serotypes in Salmonella. It may be helpful to developing a rapid, sensitive, accurate and PCR-based method to detect Salmonella strains with serotype H-1-d.
探索沙门氏菌菌株中鞭毛蛋白C的基因型与抗原血清型之间的关系。
对质粒pLS408中的沙门氏菌鞭毛蛋白C片段进行克隆、测序,并与其他菌株的相应序列进行比较。从门诊患者中分离出包括两株伤寒菌株、一株副伤寒菌株、一株肠炎沙门氏菌和一株鼠伤寒沙门氏菌在内的沙门氏菌菌株。分别从这些临床分离株中纯化基因组DNA,然后通过聚合酶链反应扩增相应的鞭毛蛋白C片段,并通过琼脂糖凝胶电泳分析扩增产物。
克隆片段包含582个核苷酸,编码质粒pLS408中沙门氏菌鞭毛蛋白C的可变区和部分保守区。与先前报道的相应序列相比,在核苷酸和氨基酸水平上与具有相同鞭毛血清型的其他菌株只有微小差异。在具有鞭毛血清型H-1-d的沙门氏菌菌株(包括慕尼黑沙门氏菌、伤寒沙门氏菌和鼠伤寒沙门氏菌)中扩增出特异性PCR产物,但在副伤寒C沙门氏菌或肠炎沙门氏菌菌株中未扩增出。
在本实验中,在鞭毛蛋白C中央可变区可发现核苷酸序列的特异性,如同在沙门氏菌鞭毛血清型中存在的那样。这可能有助于开发一种基于PCR的快速、灵敏、准确的方法来检测血清型为H-1-d的沙门氏菌菌株。