Salmonella is one of the major pathogenic bacteria present in contaminated water. 16-23S rRNA spacer region has been reported to be polymorphic at serovar level in Salmonella. Salmonella isolates obtained from Ganges ...Salmonella is one of the major pathogenic bacteria present in contaminated water. 16-23S rRNA spacer region has been reported to be polymorphic at serovar level in Salmonella. Salmonella isolates obtained from Ganges river water were studied for 16-23S rRNA spacer region polymorphism. Thirty three isolates belonging to eight serovars (S. Typhimurium, S. Abuja, S. Pantypridd, S. Lagos, S. Chinkual, S. Zwickau, S. Goldenberg and S. Oritamerin) were studied for the polymorphism. Out of 33 isolates, 15 different profiles were observed no serovar specific profile. Our findings indicate that 16-23S rRNA spacer region is not specific at serovar level, but can be used for differentiation of different Salmonella isolates.展开更多
Objective To set up a rapid and simple method for identificating bacteria by 16S 23SrDNA intergenic spacer regions (ISRs) Methods Polymorphic products of PCR from ISRs were selected on agarose gel and sequenced ...Objective To set up a rapid and simple method for identificating bacteria by 16S 23SrDNA intergenic spacer regions (ISRs) Methods Polymorphic products of PCR from ISRs were selected on agarose gel and sequenced directly using purified fragments by excising the gel without cloning Nucleotide sequences were compared with GenBank databases and analyzed by DNAMAN program Results There was only a single product in streptococcus genus after PCR amplification of 16S 23SrDNA ISRs Five streptococcal species were obtained from 7 strains of streptococcus Two major amplicons were consistently generated for 8 strains of Haemophilus influenzae (H influenzae) The sequence data showed that they all belonged to H influenzae type b on GenBank databases Conclusion PCR and direct sequencing of 16S 23SrDNA ISRs were very successful methods for bacterial species identification展开更多
根据GenBank中乳酸杆菌16 S^23 S rRNA基因间沉默区序列设计引物,进一步鉴定猪源乳酸杆菌分离株HZ521;并通过体外试验,以嗜酸乳酸杆菌ATCC 4356为参考菌株,探讨HZ521株的益生素作用。部分16 S^23 S rRNA基因序列同源性分析结果表明,该...根据GenBank中乳酸杆菌16 S^23 S rRNA基因间沉默区序列设计引物,进一步鉴定猪源乳酸杆菌分离株HZ521;并通过体外试验,以嗜酸乳酸杆菌ATCC 4356为参考菌株,探讨HZ521株的益生素作用。部分16 S^23 S rRNA基因序列同源性分析结果表明,该分离株属于瑞士乳酸杆菌。HZ521株具有较强的产酸能力,能耐受强酸,对Hela细胞的黏附率为87.2%,显著高于ATCC4356株(P<0.01)。表明瑞士乳酸杆菌HZ521株具有益生素特性,可作为肠道益生素的候选菌株。展开更多
To establish a rapid identification method for common pathogenic bacteria on the basis of molecular biology and to construct a preliminary Polymerase Chain Reaction-Capillary Electrophoresis - Restriction Fragment Len...To establish a rapid identification method for common pathogenic bacteria on the basis of molecular biology and to construct a preliminary Polymerase Chain Reaction-Capillary Electrophoresis - Restriction Fragment Length Polymorphism (PCR-CE-RFLP) database of bacteria isolated from clinical specimens frequently, 183 strains collected from clinical samples belonging to 12 genera and 19 species whose biochemical characterizations corresponded to the typical ones were examined. The genomic DNAs were amplified by two pairs of fluorescence labeled primers aiming at 16S rRNA gene and 16S-23S rRNA spacer region gene respectively at the same time. PCR products were then digested by restriction endonuclease HaeⅢ incompletely before taking capillary electrophoresis. The results with the PCR-CE-RFLP patterns of 16S rRNA genes were just alike within some genera, but when it comes to 16S-23S rRNA spacer region genes, each bacterium showed a unique pattern, which can be distinguished from each other easily. It seems that PCR-CE-RFLP patterns of 16S rRNA gene could only be used to classify the bacteria into family level, whereas the data of 16S-23S rRNA spacer region gene could be utilized to identify the whole microorganisms as precisely as the species level. In spite of the data of the spacer region gene alone can be sufficiently to verify the whole bacteria, we insist that the 16S rRNA gene could be of some assistant in case that there should be lots of families of bacteria, in which some similar ones, with the same RFLP data of 16S-23S rRNA spacer region gene, may coexist. This study proves that the utility of PCR-CE-RFLP is a convenient, rapid method to identify pathogenic bacteria, and is also a quick diagnosis measure for application to clinical use.展开更多
To study the genetic characterization of four strains of Borrelia burgdorferi isolated in China. PCR technique was used to amplify the 5S-23S rRNA intergenic spacer DNA from the whole cellular DNA of isolated GXLD-4, ...To study the genetic characterization of four strains of Borrelia burgdorferi isolated in China. PCR technique was used to amplify the 5S-23S rRNA intergenic spacer DNA from the whole cellular DNA of isolated GXLD-4, 9, 18 and Chang 14, and then the amplified products were cloned into plasmid pGEM-T Easy and sequenced. It was found that the 5S-23S rRNA intergenic spacer DNA of the four isolates was 242?bp, revealing the nucleotide sequence identity of more than 99%. The four isolates had higher sequence identify with Borrelia valaisiana than with other genetic groups. These four isolates most likely belong to Borrelia valaisiana genomic group.展开更多
文摘Salmonella is one of the major pathogenic bacteria present in contaminated water. 16-23S rRNA spacer region has been reported to be polymorphic at serovar level in Salmonella. Salmonella isolates obtained from Ganges river water were studied for 16-23S rRNA spacer region polymorphism. Thirty three isolates belonging to eight serovars (S. Typhimurium, S. Abuja, S. Pantypridd, S. Lagos, S. Chinkual, S. Zwickau, S. Goldenberg and S. Oritamerin) were studied for the polymorphism. Out of 33 isolates, 15 different profiles were observed no serovar specific profile. Our findings indicate that 16-23S rRNA spacer region is not specific at serovar level, but can be used for differentiation of different Salmonella isolates.
文摘Objective To set up a rapid and simple method for identificating bacteria by 16S 23SrDNA intergenic spacer regions (ISRs) Methods Polymorphic products of PCR from ISRs were selected on agarose gel and sequenced directly using purified fragments by excising the gel without cloning Nucleotide sequences were compared with GenBank databases and analyzed by DNAMAN program Results There was only a single product in streptococcus genus after PCR amplification of 16S 23SrDNA ISRs Five streptococcal species were obtained from 7 strains of streptococcus Two major amplicons were consistently generated for 8 strains of Haemophilus influenzae (H influenzae) The sequence data showed that they all belonged to H influenzae type b on GenBank databases Conclusion PCR and direct sequencing of 16S 23SrDNA ISRs were very successful methods for bacterial species identification
文摘根据GenBank中乳酸杆菌16 S^23 S rRNA基因间沉默区序列设计引物,进一步鉴定猪源乳酸杆菌分离株HZ521;并通过体外试验,以嗜酸乳酸杆菌ATCC 4356为参考菌株,探讨HZ521株的益生素作用。部分16 S^23 S rRNA基因序列同源性分析结果表明,该分离株属于瑞士乳酸杆菌。HZ521株具有较强的产酸能力,能耐受强酸,对Hela细胞的黏附率为87.2%,显著高于ATCC4356株(P<0.01)。表明瑞士乳酸杆菌HZ521株具有益生素特性,可作为肠道益生素的候选菌株。
文摘To establish a rapid identification method for common pathogenic bacteria on the basis of molecular biology and to construct a preliminary Polymerase Chain Reaction-Capillary Electrophoresis - Restriction Fragment Length Polymorphism (PCR-CE-RFLP) database of bacteria isolated from clinical specimens frequently, 183 strains collected from clinical samples belonging to 12 genera and 19 species whose biochemical characterizations corresponded to the typical ones were examined. The genomic DNAs were amplified by two pairs of fluorescence labeled primers aiming at 16S rRNA gene and 16S-23S rRNA spacer region gene respectively at the same time. PCR products were then digested by restriction endonuclease HaeⅢ incompletely before taking capillary electrophoresis. The results with the PCR-CE-RFLP patterns of 16S rRNA genes were just alike within some genera, but when it comes to 16S-23S rRNA spacer region genes, each bacterium showed a unique pattern, which can be distinguished from each other easily. It seems that PCR-CE-RFLP patterns of 16S rRNA gene could only be used to classify the bacteria into family level, whereas the data of 16S-23S rRNA spacer region gene could be utilized to identify the whole microorganisms as precisely as the species level. In spite of the data of the spacer region gene alone can be sufficiently to verify the whole bacteria, we insist that the 16S rRNA gene could be of some assistant in case that there should be lots of families of bacteria, in which some similar ones, with the same RFLP data of 16S-23S rRNA spacer region gene, may coexist. This study proves that the utility of PCR-CE-RFLP is a convenient, rapid method to identify pathogenic bacteria, and is also a quick diagnosis measure for application to clinical use.
文摘To study the genetic characterization of four strains of Borrelia burgdorferi isolated in China. PCR technique was used to amplify the 5S-23S rRNA intergenic spacer DNA from the whole cellular DNA of isolated GXLD-4, 9, 18 and Chang 14, and then the amplified products were cloned into plasmid pGEM-T Easy and sequenced. It was found that the 5S-23S rRNA intergenic spacer DNA of the four isolates was 242?bp, revealing the nucleotide sequence identity of more than 99%. The four isolates had higher sequence identify with Borrelia valaisiana than with other genetic groups. These four isolates most likely belong to Borrelia valaisiana genomic group.
