EurekaMag.com logo
+ Site Statistics
References:
53,869,633
Abstracts:
29,686,251
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Hemadsorption and elisa nitrocellulose replica methods for identification of colonization factor antigen positive escherichia coli colonies and for isolation of colonization factor antigen negative mutants



Hemadsorption and elisa nitrocellulose replica methods for identification of colonization factor antigen positive escherichia coli colonies and for isolation of colonization factor antigen negative mutants



Journal of Clinical Microbiology 24(4): 615-619



Methods were developed that allow demonstration of individual colonies carrying colonization factor antigen (CFA) I or CFA/II or E8775-type antigen in mixed bacterial cultures on solid media. These methods are based on mannose-resistant hemadsorption or CFA enzyme-linked immunosorbent assay (ELISA) on nitrocellulose replicas of the cultures allowing simultaneous analysis of up to 200 colonies per plate. The sensitivity and specificity of the CFA ELISA nitrocellulose replica method were 97 and 99%, respectively, for CFA/I-carrying colonies and 99 and 100% for CFA/II-positive colonies; corresponding figures for the quicker and simpler hemadsorption modification were somewhat lower. Both methods seem to be useful for studying excretion of CFA-carrying bacteria in feces, as indicated by studies in rabbits infected with enterotoxin-producing Escherichia coli in a nonligated-intestine model. By initially absorbing CFA-carrying bacteria on erythrocytes and then performing nitrocellulose replicas of agar colonies of the nonabsorbed bacteria, CFA-deficient mutants could be identified by the hemadsorption method, as well as by the CFA ELISA. Treatment of CFA-carrying bacteria with antiserum against CFA and complement also resulted in enrichment of spontaneous CFA-deficient mutants that could be identified by the replica methods. Several stable CFA-deficient mutants from enterotoxin-producing E. coli carrying CFA/I, CFA/II, or E8775 were isolated by these approaches.

(PDF emailed within 1 workday: $29.90)

Accession: 005553628

Download citation: RISBibTeXText



Related references

Hemadsorption and enzyme-linked immunosorbent assay nitrocellulose replica methods for identification of colonization factor antigen (CFA)-positive Escherichia coli colonies and for isolation of CFA-negative mutants. Journal of Clinical Microbiology 24(4): 615-619, 1986

Use of nonradioactive DNA hybridization for identification of enterotoxigenic Escherichia coli harboring genes for colonization factor antigen I, coli surface antigen 4, or putative colonization factor O166. Journal of Clinical Microbiology 30(7): 1823-1828, 1992

Genetic relationship of putative colonization factor O166 to colonization factor antigen I and coli surface antigen 4 of enterotoxigenic Escherichia coli. Infection and Immunity 60(9): 3799-3806, 1992

Genetic relationship of putative colonization factor o166 to colonization factor antigen i and coil surface antigen i and coli surface antigen 4 of enterotoxigenic escherichia coli. Infection & Immunity 60(9): 3799-3806, 1992

Positive regulation of colonization factor antigen I (CFA/I) production by enterotoxigenic Escherichia coli producing the colonization factors CS5, CS6, CS7, CS17, PCFO9, PCFO159:H4 and PCFO166. Journal of General Microbiology 137(8): 1963-1970, 1991

Re challenge of volunteers with entero toxigenic st plus lt plus colonization factor antigen type 1 escherichia coli evidence for immunity induced by colonization factor antigen type 1. Clinical Research 26(6): 771A, 1978

Production and characterization of monoclonal antibodies to a pilus colonization factor (colonization factor antigen III) of human enterotoxigenic Escherichia coli. Infection and Immunity 57(11): 3452-3457, 1989

Fimbriae of colonization factor antigen ii positive enterotoxigenic escherichia coli. Pediatric Research 18(10): 1061, 1984

Roles of different coli surface antigens of colonization factor antigen II in colonization by and protective immunogenicity of enterotoxigenic Escherichia coli in rabbits. Infection and Immunity 58(2): 341-346, 1990

Detection of colonization factor antigen I-positive enterotoxigenic Escherichia coli with a cloned polynucleotide probe. Journal of Clinical Microbiology 28(10): 2264-2268, 1990

Infection with colonization factor antigen I-expressing enterotoxigenic Escherichia coli boosts antibody responses against heterologous colonization factors in primed subjects. Epidemiology & Infection. 119(3): 391-393,., 1997

Infection with colonization factor antigen I-expressing enterotoxigenic Escherichia coli boosts antibody responses against heterologous colonization factors in primed subjects. Epidemiology and Infection 119(3): 391-393, 1998

Identification of Novel Components Influencing Colonization Factor Antigen I Expression in Enterotoxigenic Escherichia coli. Plos One 10(10): E0141469-E0141469, 2016

Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans. Infection and Immunity 44(2): 409-420, 1984

Induction of colonization factor antigen I and coli surface antigen 4 of enterotoxigenic Escherichia coli Relevance for vaccine production. Vaccine 11(2): 221-226, 1993