AIM: To prepare a kind of magnetic iron-dextran nanopartides that was coated with anti-E.coli O157:H7 IgG, analyze its application conditions, and try to use it to isolate E.coli O157:H7 from foods. METHODS: Magnetic ...AIM: To prepare a kind of magnetic iron-dextran nanopartides that was coated with anti-E.coli O157:H7 IgG, analyze its application conditions, and try to use it to isolate E.coli O157:H7 from foods. METHODS: Magnetic iron-dextran nanopartides were prepared by the reaction of a mixture of ferric and ferrous ions with dextran polymers under alkaline conditions. The particles were coated with antiserum against E.coli O157: H7 by the periodate oxidation-borohydride reduction procedure. The oxidation time, amount of antibody coating the particles, amount of nanoparticles, incubation time and isolation time were varied to determine their effects on recovery of the organisms. Finally, the optimum conditions for isolating E.coli O157:H7 from food samples were established. RESULTS: E.coli O157:H7 can be isolated from samples within 15 min with the sensitivity of 101 CFU/mL or even less. In the presence of 108 CFU/mL of other organisms, the sensitivity is 101-102 CFU/mL. Nonspecific binding of other bacteria to the particles was not observed. Two and a half hours of enrichment is enough for the particles to detect the target from the food samples inoculated with 1 CFU/g. CONCLUSION: Isolation of target bacteria by immuno magnetic nanoparticles is an efficient method with high sensitivity and specificity. The technique is so simple that it can be operated in lab and field even by untrained personnel.展开更多
Objective: To evaluate the zoonotic potency of Escherichia coli O157:H7 through arbitrarily primed-PCR(AP-PCR) methods as one of the DNA fingerprinting methods.Methods: A total of 14 isolates consisted of 11 isolates ...Objective: To evaluate the zoonotic potency of Escherichia coli O157:H7 through arbitrarily primed-PCR(AP-PCR) methods as one of the DNA fingerprinting methods.Methods: A total of 14 isolates consisted of 11 isolates originated from human feces with renal failure symptoms, 2 isolates originated from cattle feces, and 1 control isolate were used in this study. DNA of each isolate was extracted, and their pro files were studied by using AP-PCR method with M13 F and M13 R arbitrary primers.Results: The results founded that all of 14 isolates had similarity range from 54.6% to88.5%. Isolates KL-106(3) and KL-55(6) originated from humans showed the degree of similarity with isolates SM-25(1) and SM-7(1) originated from cattle as high as 85% and77%, respectively.Conclusions: The high degree of similarity between isolates originated from cattle and human indicated the high potency of zoonoses. The results also concluded AP-PCR method as a brie fly fingerprinting method in order to trace the epidemiological of E. coli O157:H7.展开更多
The genome of the enterohemorrhagic Escherichia coli O157∶H7 EDL933 contains 177 “O”-islands (OIs). To study their potential contribution to the O157-specific pathogenicity, we surveyed the distribution of 22 OIs b...The genome of the enterohemorrhagic Escherichia coli O157∶H7 EDL933 contains 177 “O”-islands (OIs). To study their potential contribution to the O157-specific pathogenicity, we surveyed the distribution of 22 OIs by PCR and DNA hybridization in 17 isolates of Shiga toxin producing (Stx-positive) E.coli O157∶H7, and compared with their distribution in 21 isolates of Stx-negative E.coli O157 and 21 isolates of non-O157 enteric pathogens. Fourteen of 22 OIs were present in non-O157 entericpathogens analyzed. Eight of 22 OIs were found only in the 17 Shiga toxin- (Stx) positive E.coli O157∶H7 isolates, but they were absent from the 21 Stx-negative E.coli O157∶NM and O157︰Hund isolates tested. Among the 8 OIs, only OI43 or OI48 were exclusively detected in Stx-positive E.coli O157∶H7, absent from neither of Stx-negative E.coli O157 and non-O157 enteric pathogens, such as Salmonella, Shigella, Citrobacter, Vibrio cholera, enteropathogenic E.coli (EPEC), enteroadherent E.coli (EAEC), enteroinvasive E.coli (EIEC) and enterotoxingenic E.coli (ETEC). The OI43 and OI48 are 83 kb in size and identical in DNA sequences, which encode genes for urease, tellurite resistance and adherence. By analyzing their junction genes with PCR and DNA hybridization, we found that 21 Chinese isolates have OI48 only. However, for 7 Japanese patient isolates, 4 have OI43 and 3 have OI48; for American isolates, 2 have both of OI43 and OI48, 2 have OI48 only. These data confirmed the highly plasticity of the pathogenic E.coli genome. The unique presence of OI43/OI48 in Stx-positive E.coli O157∶H7 denotes its critical role in the pathogenicity specific to this pathogen.展开更多
基金Supported by the National High-technology Research and Development Program of China (863 Program), No. 2003AA302260
文摘AIM: To prepare a kind of magnetic iron-dextran nanopartides that was coated with anti-E.coli O157:H7 IgG, analyze its application conditions, and try to use it to isolate E.coli O157:H7 from foods. METHODS: Magnetic iron-dextran nanopartides were prepared by the reaction of a mixture of ferric and ferrous ions with dextran polymers under alkaline conditions. The particles were coated with antiserum against E.coli O157: H7 by the periodate oxidation-borohydride reduction procedure. The oxidation time, amount of antibody coating the particles, amount of nanoparticles, incubation time and isolation time were varied to determine their effects on recovery of the organisms. Finally, the optimum conditions for isolating E.coli O157:H7 from food samples were established. RESULTS: E.coli O157:H7 can be isolated from samples within 15 min with the sensitivity of 101 CFU/mL or even less. In the presence of 108 CFU/mL of other organisms, the sensitivity is 101-102 CFU/mL. Nonspecific binding of other bacteria to the particles was not observed. Two and a half hours of enrichment is enough for the particles to detect the target from the food samples inoculated with 1 CFU/g. CONCLUSION: Isolation of target bacteria by immuno magnetic nanoparticles is an efficient method with high sensitivity and specificity. The technique is so simple that it can be operated in lab and field even by untrained personnel.
基金Supported by the Directorate of Research and Community Services,Directorate General of Higher Education through Udayana Research Grants with contract No.21.34/UN14/SBRC/2012
文摘Objective: To evaluate the zoonotic potency of Escherichia coli O157:H7 through arbitrarily primed-PCR(AP-PCR) methods as one of the DNA fingerprinting methods.Methods: A total of 14 isolates consisted of 11 isolates originated from human feces with renal failure symptoms, 2 isolates originated from cattle feces, and 1 control isolate were used in this study. DNA of each isolate was extracted, and their pro files were studied by using AP-PCR method with M13 F and M13 R arbitrary primers.Results: The results founded that all of 14 isolates had similarity range from 54.6% to88.5%. Isolates KL-106(3) and KL-55(6) originated from humans showed the degree of similarity with isolates SM-25(1) and SM-7(1) originated from cattle as high as 85% and77%, respectively.Conclusions: The high degree of similarity between isolates originated from cattle and human indicated the high potency of zoonoses. The results also concluded AP-PCR method as a brie fly fingerprinting method in order to trace the epidemiological of E. coli O157:H7.
基金This work was supported by the Basic Research Program from Ministry of Science and Technology,China (G1999054101 to J.Xu.) and PRA program from AFCRST (B99 03) to LFW
文摘The genome of the enterohemorrhagic Escherichia coli O157∶H7 EDL933 contains 177 “O”-islands (OIs). To study their potential contribution to the O157-specific pathogenicity, we surveyed the distribution of 22 OIs by PCR and DNA hybridization in 17 isolates of Shiga toxin producing (Stx-positive) E.coli O157∶H7, and compared with their distribution in 21 isolates of Stx-negative E.coli O157 and 21 isolates of non-O157 enteric pathogens. Fourteen of 22 OIs were present in non-O157 entericpathogens analyzed. Eight of 22 OIs were found only in the 17 Shiga toxin- (Stx) positive E.coli O157∶H7 isolates, but they were absent from the 21 Stx-negative E.coli O157∶NM and O157︰Hund isolates tested. Among the 8 OIs, only OI43 or OI48 were exclusively detected in Stx-positive E.coli O157∶H7, absent from neither of Stx-negative E.coli O157 and non-O157 enteric pathogens, such as Salmonella, Shigella, Citrobacter, Vibrio cholera, enteropathogenic E.coli (EPEC), enteroadherent E.coli (EAEC), enteroinvasive E.coli (EIEC) and enterotoxingenic E.coli (ETEC). The OI43 and OI48 are 83 kb in size and identical in DNA sequences, which encode genes for urease, tellurite resistance and adherence. By analyzing their junction genes with PCR and DNA hybridization, we found that 21 Chinese isolates have OI48 only. However, for 7 Japanese patient isolates, 4 have OI43 and 3 have OI48; for American isolates, 2 have both of OI43 and OI48, 2 have OI48 only. These data confirmed the highly plasticity of the pathogenic E.coli genome. The unique presence of OI43/OI48 in Stx-positive E.coli O157∶H7 denotes its critical role in the pathogenicity specific to this pathogen.
