New fimbrial antigenic type (E8775) that may represent a colonization factor in enterotoxigenic Escherichia coli in humans.

An enterotoxigenic strain of Escherichia coli O25:H42 (strain E8775), isolated from a patient in Bangladesh with diarrhea, caused mannose-resistant hemagglutination (MRHA) of human and bovine erythrocytes. The strain did not show slide agglutination or immunodiffusion precipitin lines with antiserum specific for the colonization factor antigen CFA/I or CFA/II. A variant E. coli strain, E8775-B, did not cause MRHA or produce enterotoxin. Electron microscopy revealed the presence of fimbriae on the surface of strain E8775 but not strain E8775-B. When strain E8775 was grown at 22 degrees C, it became MRHA negative and fimbriae were absent. An antiserum prepared against strain E8775 was absorbed with strain E8775-B to make an antiserum specific for the fimbrial antigen. Using this absorbed antiserum, we found the fimbrial antigen in 48 of 742 enterotoxigenic E. coli strains. The 48 strains belonged to serogroups O25, O115, and O167. It is suggested by analogy to the properties of previously described colonization factors that these fimbriae may play a part in the colonization of the intestinal epithelium.     

New Surface-Associated Heat-Labile Colonization Factor Antigen (CFA/II) Produced by Enterotoxigenic Escherichia coli of Serogroups O6 and O8

Enterotoxigenic Escherichia coli (ETEC) belonging to serogroups O6 and O8 do not possess the H-10407-type colonization factor antigen (CFA/I). However, these frequently isolated ETEC were found to possess a second and distinct heat-labile surface-associated colonization factor antigen, termed CFA/II. Whereas CFA/I mediates mannose-resistant hemagglutination of human group A erythrocytes, CFA/II does not. CFA/II mediates mannose-resistant hemagglutination of bovine erythrocytes, and mannose-resistant hemagglutination is rapid only at reduced temperature (4 degrees C). Because CFA/II, like CFA/I, is spontaneously lost by many ETEC isolates in the laboratory, it was possible to produce specific anti-CFA/II serum by preparing antiserum against living cells of a prototype strain (PB-176) and adsorbing this serum with living and heat-treated cells of its CFA/II-negative derivative strain PB-176-P. This serum, which neutralized the colonization factor activity of CFA/II-positive strains in infant rabbits, was employed to confirm the presence of CFA/II on ETEC which exhibited mannose-resistant hemagglutination of bovine but not human erythrocytes. CFA/II, like CFA/I, mediates adherence of the bacteria to the mucosal surface of the small intestine, as demonstrated by indirect immunofluorescence. CFA/II appears to be an important virulence factor for humans since CFA/II-positive ETEC are frequently isolated from diarrhea cases, particularly travelers' diarrhea, in Mexico; these ETEC were not uncommon in a collection of isolates from Bangladesh. The O6:H16 strain of ETEC responsible for an outbreak of diarrhea in the United States was also shown to be CFA/II positive. CFA/I and CFA/II were never found on the same serotypes of ETEC, but 98% of the heat-stable and heat-labile enterotoxin-producing ETEC belonging to the frequently isolated serogroups O6, O8, O15, O25, O63, and O78 were positive for either CFA/I or CFA/II.     

Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans.

Enterotoxigenic Escherichia coli (ETEC) of serotype O6:H16, biotype A, bearing colonization factor antigen II (CFA/II) possesses two distinct coli surface antigens, CS1 and CS3, whereas CFA/II-positive ETEC of serotype O8:H9 manifests only CS3. CS1 has been shown to be fimbrial in nature, but heretofore the morphology of CS3 has not been described. Accordingly, by immune electron microscopy we investigated the morphological characteristics of CS3 on bacterial cells and after purification. CS3 was found to consist of thin (2-nm), flexible, wiry, "fibrillar" fimbriae, visible both on bacteria (O6:H16, biotype A, and O8:H9 strains) and in the pure state. In contrast, CS1 exists as wider (6-nm), rigid fimbriae on the surface of O6:H16, biotype A, strains. By the use of antisera to CS1 and CS3 in immune electron microscopy, immunodiffusion in gel, and immunoblotting techniques, CS1 and CS3 were found to be immunologically as well as morphologically distinct. Six of nine volunteers who developed diarrhea after challenge with an O139:H28 ETEC strain bearing CS1 and CS3 had significant serological rises to purified CS1 and CS3 antigens, suggesting that both antigens are elaborated in vivo, play a role in pathogenesis, and stimulate an immune response.     

Colonization factors of enterotoxigenic Escherichia coli isolated from children with diarrhea in Argentina.

A prospective study was performed to evaluate the presence of colonization factor antigens (CFAs) in enterotoxigenic Escherichia coli (ETEC) strains isolated from 1,211 children with diarrhea in Argentina. One hundred nine ETEC strains that were isolated from seven different laboratories in various regions of the country were tested for CFAs by using monoclonal antibodies against CFA/I and E. coli surface antigens CS1, CS2, and CS3 of CFA/II and CS4 and CS5 of CFA/IV; a polyclonal antiserum against CS6 was used. The CFAs searched for were found in 52% of the ETEC strains: 23% of the strains carried CFA/I, 17% carried CFA/IV, and 12% carried CFA/II. All of the CFA/I strains produced heat-stable enterotoxin, and several of them were of the prevalent serotypes O153:H45 and O78:H12. Among the 19 strains expressing CFA/IV, 16 expressed CS5 and CS6 and produced the heat-stable enterotoxin and most were of serotype O128:H21; the remaining 3 strains produced CS6 only. No ETEC strains expressing CS4 were found. Most (11 of 13) of the CFA/II-carrying ETEC strains expressed CS1 and CS3, and 10 of them were of the O6:K15:H16 serotype and produced both heat-labile and heat-stable toxins. As many as 24 of the 109 CFA-negative ETEC strains gave mannose-resistant hemagglutination with erythrocytes from different species; 4 strains had high surface hydrophobicity, suggesting the presence of additional, as yet undefined, colonization factors in up to 25% of the ETEC isolates.     

The occurence of colonisation factors (CFA/I,CFA/II and E8775) in enterotoxigenicEscherichia coli from various countries in South East Asia

Four hundred and fifty-eight enterotoxigenic strains ofEscherichia coli (ETEC) were examined for the presence of colonisation factor antigens (CFA) I and II, and the putative colonisation factor, E8775, using an immunodiffusion technique with specific antisera. The ETEC strains had been isolated in Thailand, Bangladesh and from travellers returning to Japan from abroad. Approximately 14% of the ETEC strains possessed CFA/I and a further 13% of the strains possessed CFA/II. The E8775 antigen was found on 5% of the strains. CFA/I was found on strains of the serogroups 04, 015, 063, 078, 090, 0110, 0126, 0128, 0153 and 0? CFA/II was found on strains of the serogroups 06, 08, 09, 078, 0115, 0139, 0? and 0 rough. The E8775 antigen was found on strains of the serogroups 025, 0115 and 0167. The results of this study emphasise the need to continue the search for other mechanisms of adhesion used by ETEC strains, and in particular strains of the serogroups 027, 034, 0148 and 0166.     

Hemagglutination patterns of enterotoxigenic and enteropathogenic Escherichia coli determined with human, bovine, chicken, and guinea pig erythrocytes in the presence and absence of mannose.

A hemagglutination (HA)-typing system has been developed for the presumptive identification of enterotoxigenic Escherichia coli (ETEC) possessing the colonization factor antigens (CFA) CFA/I or CFA/II. E. coli isolates are grown on CFA agar and tested for mannose-sensitive (MS) or mannose-resistant (MR) HA of human, bovine, chicken, and guinea pig erythrocytes. CFA/I-positive ETEC exhibit MRHA with human, bovine, and chicken erythrocytes, but no HA with guinea pig erythrocytes. CFA/II-positive ETEC produce HA (MRHA) only with bovine and chicken erythrocytes. Common pili appear to be the primary MS-hemagglutinin of E. coli because the prototype strain K-12 exhibits HA (MSHA) with all but bovine erythrocytes. However, only 6.6% (23 of 351) of E. coli belonging to the classical enteropathogenic E. coli serogroups (EPEC) possessed the same HA pattern as strain K-12; 42% of the EPEC cultures (146 of 351) were similar to K-12 in producing MSHA with chicken and guinea pig erythrocytes and no HA with bovine erythrocytes, but different in that these produced either no HA or MRHA with human erythrocytes. These EPEC-associated HA patterns were assigned to a separate category, termed HA type III. Non-EPEC serogroups associated with sporadic diarrhea (i.e., the facultatively enteropathogenic E. coli, or FEEC) and 41% (19 of 46) of available Salmonella isolates also produced HA type III patterns. This observation is of considerable interest because many FEEC possess somatic antigens cross-reactive with Salmonella. Although the biochemical basis for this result has not been established, the data reported herein suggest a relationship between the HA type III phenotype and virulence (enteropathogenicity) in both the EPEC and FEEC serogroups. We propose that HA typing be used in conjunction with serotyping of E. coli to determine the degree of association of HA type III E. coli with sporadic diarrhea in infants and young children.     

Plasmids that code for production of colonization factor antigen II and enterotoxin production in strains of Escherichia coli.

Similar plasmids that code for production of colonization factor antigen II and enterotoxins were mobilized into Escherichia coli K-12 from strains of E. coli O6.H16 biotype B or C that expressed colonization factor antigen II components CS2 and CS3. Further transfer experiments demonstrated that there was production of CS2 and CS3 in an O6.H16 biotype C strain, but there was expression of only CS3 in other serogroups and in E. coli K-12.     

Hemagglutination by Escherichia coli in septicemia and urinary tract infections.

The agglutination of erythrocytes from various animal species by Escherichia coli was studied. The 405 strains of E. coli were isolated from urine in patients with urinary tract infections, from blood in septicemic patients, or from feces in persons without intestinal or urinary disorders. In urinary tract infections, d-mannose-resistant agglutination (MRHA) of human erythrocytes was the most common finding (23% of the strains). The highest frequency of mannose-sensitive hemagglutination (MSHA) attributed to type I (common type) pili occurred with guinea pig erythrocytes (11.5%). Of the 78 E. coli strains isolated from blood cultures, 11 (14%) produced MRHA of human erythrocytes and only one gave MSHA. In the stool cultures, only 1 of 170 E. coli strains was MSHA reacting, whereas 28 strains (16.5%) showed MRHA of human erythrocytes. No MRHA strain reacted with antiserum against colonization factor antigen (CFA)/I of pilus nature in enterotoxigenic human E. coli strains (O78:H12). MRHA of bovine erythrocytes, reputedly typical of enterotoxigenic E. coli of serogroups O6 and O8, was shown by only two strains, neither of which agglutinated with CFA/II antiserum. The most common hemagglutinating pattern of E. coli from urine and blood thus was MRHA for human erythrocytes. This agglutination may have been caused by pili or other surface properties of one or more serotypes. These may represent a new class of colonization-promoting antigens (adhesins).     

Monoclonal antibodies against the different subcomponents of colonization factor antigen II of enterotoxigenic Escherichia coli.

Monoclonal antibodies (MAbs) against the different coli surface antigens CS1, CS2, and CS3 of colonization factor antigen II (CFA/II) of enterotoxigenic Escherichia coli (ETEC) were generated by fusing F/O myeloma cells with spleen cells from BALB/c mice immunized with different preparations of purified CFA/II. Five hybrids that produced antibodies specific for CS1, CS2, or CS3 in high titer were cloned and propagated. All the anti-CS MAbs were of the immunoglobulin G1 isotype, and all gave single precipitation lines in immunodiffusion tests when reacting with CFA/II-positive E. coli extracts containing the corresponding CS factor. The binding of all the MAbs to solid-phase-bound CFA/II could be completely inhibited by purified CFA/II containing the corresponding CS factor. However, whereas one MAb against CS3 was inhibited by all of 18 different CFA/II-positive strains tested, another anti-CS3 MAb was inhibited by bacteria expressing CS1 and CS3 (CS1 + CS3 strains) or CS3 alone but not by CS2 + CS3 strains, suggesting antigenic differences in CS3 when expressed by different strains. Use of the anti-CS MAbs in slide agglutination, immunodiffusion, or a CFA inhibition enzyme-linked immunosorbent assay revealed differences in the relative distribution of the various CS factors of CFA/II in clinical ETEC isolates from different geographic areas. By using the anti-CS MAbs in an enzyme-linked immunosorbent assay-nitrocellulose replica method, CFA/II-positive colonies could be detected in stool cultures from infected animals without prior isolation of the ETEC organisms.     

Prevalence of the enterotoxigenic Escherichia coli colonization factors CFA/I, CFA/II and E8775 isolated in the South Pacific (New Caledonia, Republic of Vanuatu and Wallis and Futuna)

We tested the expression of adherence properties of enterotoxigenic Escherichia coli (ETEC) strains isolated in New-Caledonia, Vanuatu and Wallis and Futuna by examining for the presence of colonization factor E8775 using an agglutination test and an immuno-diffusion technique with specific antisera. Approximately 19% of ETEC strains possessed CFA/I and 21% a CFA/II. The E8775 antigen was found on 1.8% of the strains. This last factor was found on strains of the serogroup 025 from Vanuatu. Two strains 078 usually CFA/I+ possessed a CFA/II and three strains of the serogroup 0126 possessed a CFA/I. The results of this study emphasis the need to continue the search for other mechanisms of adhesion used by ETEC strains without any of the three factors of colonization.     

Synergistic protective effect of antibodies against Escherichia coli enterotoxin and colonization factor antigens.

We studied the ability of antisera against different Escherichia coli surface antigens, both alone and in combination with anti-enterotoxin, to decrease fluid secretion induced by intestinal challenge with enterotoxigenic E. coli in rabbits. Antiserum against lipopolysaccharide protected significantly against O group homologous bacteria. Monospecific antisera against pilus-associated colonization factor antigens CFA/I and CFA/II were also effective, giving highly significant protection against enterotoxigenic E. coli strains bearing the corresponding colonization factor antigens. Protection was also observed with Fab fragments of the CFA/I antibodies. Addition of the anti-lipopolysaccharide serum to a protective antiserum against purified heat-labile enterotoxin resulted in an antisecretory effect which slightly exceeded the sum of the effects obtained with each preparation alone. The combination of antiserum against CFA/I or CFA/II with anti-enterotoxin gave protection that equaled the product of the effects obtained with each antiserum alone; i.e. the antisera cooperated synergistically.     

Colonization antigens and haemagglutination patterns of humanEscherichia coli

The haemagglutinating properties of 223 (35 enterotoxigenic and 188 non-enterotoxigenic)Escherichia coli strains with nine erythrocyte types were investigated; 153 strains were also tested for beta-haemolysis and colicin production and for the presence of CFA/I, CFA/II, K88 and K99 antigens. A selected group of strains was also examined by electron microscopy to determine the presence of fimbriae or fibrils and to establish the relationship between these, the haemagglutinating properties and the presence of colonization antigens. Generally, the haemagglutinating patterns yielded by the same strains grown in Mueller Hinton broth and on CFA agar differed considerably. Mannose-sensitive haemagglutinating (MSHA) patterns were more homogeneous than mannose-resistant haemagglutinating (MRHA) patterns. Forty-seven percent of the non-enterotoxigenic MRHA⁺ strains were haemolytic while only 6 % of the remaining strains were (X² correction=34.01; p<0.001). CFA/I was only detected in the four enterotoxigenic MRHA⁺ strains which were positive only with human and calf erythrocytes when grown on CFA agar. CFA/II was detected in three of a total of six enterotoxigenic strains which were MRHA⁺ only with calf erythrocytes when grown on CFA agar. K88 and K99 antigens were not detected. All strains in which bacteria with fimbriae or fibrils were observed showed haemagglutinating activity. Thus, 18 (66.7 %) of the 27 haemagglutinating strains grown on CFA agar showed fimbriae or fibrils while none of the 19 non-haemagglutinating strains did (X² correction=18.10; p<0.001).     

Genetic and molecular studies of plasmids coding for colonization factor antigen I and heat-stable enterotoxin in several escherichia coli serotypes.

Plasmids coding for colonization factor antigen I (CFA/I) and heat-stable enterotoxin (ST) were identified in 10 strains of human enterotoxigenic Escherichia coli. The strains, which belonged to serogroups O63, O114, O128, and O153, were isolated in Bangladesh, Latin America, Spain, and South Africa. Two strains produced heat-labile enterotoxin in addition to ST. CFA/I-ST plasmids were mobilized from two O128 strains into E. coli K-12 with the R factor R1-19K-. Like the prototype CFA/I-ST plasmid NTP113, mobilized previously from an E. coli O78 strain into K-12, these two plasmids were non-autotransferring. All 10 CFA/I-ST plasmids were incompatible with NTP113 and had molecular weights ranging from 59 X 10(6) to 72 X 10(6). The molecular properties of seven of these plasmids were compared with those of six CFA/I-ST plasmids previously mobilized from O78 strains from Ethiopia, South Africa, and Bangladesh and with those of one plasmid coding for CFA/I, ST and heat-labile enterotoxin from a South African strain of serogroup O63. Digestion with the restriction endonuclease HindIII showed that several plasmids had very similar fragment patterns and two were identical. Generally, a larger proportion of HindIII fragments were of common size in digests of plasmids identified in strains from related geographical areas, regardless of serogroup. However, all except one plasmid shared five or six HindIII fragments of the same size, one of which had been shown previously to be involved in CFA/I production. There was at least 90% DNA homology between CFA/I-ST plasmids with a molecular weight of about 58 X 10(6) from O78 strains from different sources. Most of the DNA sequences of these plasmids were present in a larger CFA/I-ST plasmid (72 X 10(6) from an O128 strain. The results of genetic and molecular studies suggest that CFA/I and ST production is determined by very similar plasmids in different serogroups of human enterotoxigenic E. coli from several sources.     

Identification of a new fimbrial structure in enterotoxigenic Escherichia coli (ETEC) serotype O148:H28 which adheres to human intestinal mucosa: a potentially new human ETEC colonization factor.

Three important fimbrial colonization factor antigens (CFAs) designated CFA/I, CFA/II, and E8775 were identified originally in some human enterotoxigenic Escherichia coli (ETEC) strains because of their mannose-resistant hemagglutination properties. To identify CFA, in strains lacking mannose-resistant hemagglutination properties we exploited the ability of human ETEC strains to adhere to human proximal small intestinal mucosa. ETEC strain B7A (O148:H28) was selected for study because it belongs to an epidemiologically important serotype and does not produce a known CFA, and yet it is known to be pathogenic and cause diarrheal disease in human volunteers. Results of an human enterocyte adhesion assay indicated that some bacteria in cultures of B7A produced adhesive factors. To select for such bacteria, cultured human duodenal mucosal biopsy samples were infected with B7A for up to 12 h, after which time a large percentage of the mucosal surface became colonized by bacteria. A new fimbrial structure morphologically distinct from CFA/I, CFA/II, and E8775 fimbriae and consisting of curly fibrils (approximately 3 nm in diameter) was readily identified when bacteria were subcultured from the mucosa and examined by electron microscopy. Identical fimbriae were produced by ETEC strain 1782-77 of the same serotype. Identification of these fimbriae only on bacteria subcultured from human intestinal mucosa strongly suggests that they promote mucosal adhesion of ETEC serotype O148:H28 and thus represent a potentially new human ETEC CFA.     

Relationship between enterotoxin production and serotype in enterotoxigenic Escherichia coli.

We examined the relationship between serotype and enterotoxin production in 109 enterotoxigenic Escherichia coli strains isolated from 109 patients with severe cholera-like diarrhea in Dacca, Bangladesh. Of 69 strains producing both heat-labile and heat-stable toxins, 59 (86%) belonged to the one of four O serogroups, and 56 (81%) of these strains belonged to one of six O:K:H serotypes. In contrast, 34 strains producing only heat-stable toxin were distributed among 15 O serogroups, and six strains producing only heat-labile toxin were distributed among six O serogroups. Twelve strains producing heat-labile and heat-stable toxins and five strains producing heat-stable toxin were found which had the same serotype (O78:K-:H12) and biotype. It appears that at least in one geographic setting E. coli strains producing both heat-labile and heat-stable toxins are more restricted in their O groups and O:K:H serotypes than E. coli that produce only heat-stable toxin and that certain serobiotypes may commonly include strains which produce both toxin types.     

Serological identification of pig enterotoxigenic Escherichia coli strains not belonging to the classical serotypes.

Escherichia coli strains isolated from pigs suspected to have succumbed to E. coli enterotoxicosis and not belonging to the commonly incriminated (classical) serotypes (O8:K87:K88, O45:K88, O138:K81:K88, O141:K85:K88, O147:K89:K88, O149:K91;K88, and O157:K88) were tested for enterotoxigenicity in the ligated gut test (LGT) using pig intestine. Of 202 strains tested, 54 strains belonging to 13 different O groups were positive in the LGT. Four of these strains had K88 antigen and one possessed K99 antigen. The majority of the strains was not agglutinated by any of the standard OK antisera. Four new K antigens ("K200", "K442", "K2346" and "K2347") were provisionally designated. K200 was found in pig enterotoxigenic strains belonging to O group 8 and carrying flagellar antigen H31 and in non-enterotoxigenic non-motile strains of O group 8, as well as in O group 20 strains isolated from calves succumbing to E. coli septicemia in two countries. The provisional antigen K2346 was encountered in 18 enterotoxigenic strains with various O antigens from two countries. It is proposed to include these two K antigens into the international E. coli antigens scheme. Attempts to demonstrate a common antigen in the nonclassical enterotoxigenic strains lacking K88 and K99 antigens failed.     

Production of heat-labile or heat-stable enterotoxins by strains of Escherichia coli belonging to serogroups O44, O114, and O128.

Strains of Escherichia coli which belong to enteropathogenic serogroups usually fail to produce heat-labile or heat-stable enterotoxins. However, 1 of 34 strains of E. coli O44, 9 of 45 strains of E. coli O114, and 18 of 82 strains of E. coli O128 produced heat-labile or heat-stable enterotoxins. Most enterotoxigenic isolates were from tropical or developing countries; all three enterotoxigenic strains isolated in Britain were from patients returned from abroad. Enterotoxigenic strains were of many different flagellar types. Certain enterotoxigenic strains of E. coli O114 and O128 possessed colonization factor antigen I.     

Evaluation of conventional media for detection of colonization factor antigens of enterotoxigenic Escherichia coli.

Strains of enterotoxigenic Escherichia coli producing either colonization factor antigen (CFA) I or II were tested for expression of CFA when grown on 16 different agar media by using agglutination and coagglutination with monoclonal antibodies, mannose-resistant hemagglutination, and a salt aggregation assay. CFA was detected from the CFA-positive strains when CFA agar was used, and it was also detected when other commercially available media were used, notably nutrient agar. CFA was not detected when other commercial media such as MacConkey agar were used. The use of nutrient agar with monoclonal antibody-based coagglutination reagents offers a potentially simple and rapid method for detecting E. coli which express CFA I or II.     

Attachment Factors Among Enterotoxigenic Escherichia coli from Patients with Acute Diarrhea from Diverse Geographic Areas

To cause diarrhea, enterotoxigenic Escherichia coli (ETEC) must initially colonize the small bowel. Different surface structures have been implicated in this initial attachment. Recognized attachment factors include colonization factor antigens I and II (CFA/I and CFA/II) and type I pili. Several methods of detection for each of these factors have been reported. In this study, we screened for the presence of these attachment factors among enterotoxigenic E. coli isolated from 40 patients with acute diarrhea and 40 asymptomatic control individuals and examined their ability to attach to ATCC 407 human intestinal cells in vitro. Of 40 patients with diarrhea, 16 (40%) had enterotoxigenic E. coli isolates which exhibited an attachment trait. Fourteen (35%) of these isolates demonstrated the ability to attach to ATCC 407 cells, whereas only four isolates from asymptomatic controls attached (P < 0.02). A total of 20% of the patient isolates and 7.5% of the control isolates possessed CFA/I. Only one patient isolate demonstrated CFA/II. Evidence for type I pili was found on 10% of the patient isolates and 12.5% of the control isolates. Attachment to ATCC 407 cells allowed the detection of 87.5% (14 of 16) of enterotoxigenic E. coli isolates with any type of attachment trait. Of the 14 cases demonstrating attachment ability to ATCC 407 cells, 7 did not attach in the presence of mannose. Three of these showed evidence for both CFA/I and type I pili, one showed only CFA/I, and one showed only type I pili. Two of those mannose-sensitive attaching isolates showed no other demonstrable trait. Seven patient isolates showed mannose-resistant attachment. Of these, two were classified as possessing CFA/I, and one was classified as possessing CFA/II. The four remaining isolates could not be classified into any recognized attachment factor category, suggesting that other attachment factors remain to be identified.     

Association of rns homologs with colonization factor antigens in clinical Escherichia coli isolates.

rns is a trans-acting positive regulatory factor required for expression of the colonization factor antigen II (CFA/II) antigens CS1 and CS2 (J. Caron, L. M. Coffield, and J. R. Scott, Proc. Natl. Acad. Sci. USA 86:963-967, 1989). All 35 CFA/II-positive strains hybridized with a rns gene probe, as did all 10 CFA/I strains and all 4 CS4 strains. Hybridization with rns was detected in 25% of non-enterotoxigenic Escherichia coli strains and was not detected in enteric pathogens with low G + C content.