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Biomedical subjects

H Lior

Publications and source records attributed to H Lior.

At least 55 records · Page 3Linked to original sources
PubMed1991

Application of the polymerase chain reaction (PCR) to detection of the aerolysin gene in whole cell cultures of beta-hemolytic Aeromonas hydrophila.

Oligonucleotide primers were used in a polymerase chain reaction protocol to detect the aerolysin gene in Aeromonas hydrophila. Primers detected template DNA in hemolytic, cytotoxic and enterotoxic strains of A. hydrophila and no amplification was detected with hemolytic A. sobria, non-hemolytic A. hydrophila and A. caviae strains.

Aeromonas↗
PubMed1991

Serologic evidence of Campylobacter jejuni/coli enteritis in patients with Guillain-Barré syndrome.

We performed serologic testing for Campylobacter jejuni in 17 consecutive patients with acute Guillain-Barré syndrome from the Boston, Mass area to compare the frequency of this preceding infection with the high rates reported from other areas of the world. The rate of seropositivity, 18%, was considerable, but it was lower than that reported in Australia. Moreover, all of our patients with definite serologic evidence of infection had severe enteritis before Guillain-Barré syndrome, usually with the organism cultured from stool samples. Campylobacter enteritis is an important antecedent illness for Guillain-Barré syndrome but did not precipitate the disease without enteritis.

Antibodies, Bacterial↗
PubMed1991

Distribution of Salmonella enteritidis phage types in Canada.

The distribution of Salmonella enteritidis phage types in Canada is described; 606 of 674 strains examined were of human origin. Typable strains of all sources, constituted 99.6% (671/674) of all strains examined, and were representative of 15 different phage types. Five phage types (8, 13, 4, 13a and 1) accounted for 92.4% of the total. Phage type 8 consistently showed the highest incidence in human (69.96%) and non-human (72.05%) sources and appeared to be the most common in North America. Phage type 4, the most prevalent in the UK, is infrequent in Canada (38/674). The distribution of phage types showed regional variation among infrequent phage types, whereas the common type, 8, was observed in different frequencies in all provinces. Examination of 29 outbreaks of S. enteritidis representing 254 isolates for humans revealed 5 different phage types, the highest number of outbreaks (11) were type 8. A study of these outbreaks and the animal-host-associations of the common phage types, 8 and 13, indicated that contaminated poultry appeared to be the most common source of human infection in Canada.

Animals↗
PubMed1991

Haemolytic anaemia after childhood Escherichia coli O 157 .H7 infection: are females at increased risk?

We conducted a 4-year retrospective cohort study to better define the risk of haemolytic anaemia and haemolytic uraemic syndrome (HUS) in children following sporadic gastrointestinal infection with the O 157.H7 serotype of Escherichia coli. Of the 72 children infected with this organism, 9 (12.5%) developed haemolytic anaemia, 6 of whom had HUS. No child in a cohort of 72 age-matched controls with Campylobacter jejuni gastroenteritis developed haemolytic anaemia (P = 0.003). Females had a significantly greater risk of developing haemolytic anaemia after E. coli O 157. H7 infection than did males (8/29 females v. 1/43 males; P = 0.003). In a logistic regression model, female gender emerged as the only statistically significant risk factor for haemolytic anaemia (odds ratio 3.85; 95% confidence interval 1.24-12). These results are consistent with recent reports of a moderate increase in the risk of HUS for females.

Adolescent↗
PubMed1991

Analysis of the role of flagella in the heat-labile Lior serotyping scheme of thermophilic Campylobacters by mutant allele exchange.

Flagellin mutations originally constructed in Campylobacter coli VC167 (serotype LIO8) by a gene replacement mutagenesis technique (P. Guerry, S. M. Logan, S. Thornton, and T. J. Trust, J. Bacteriol. 172:1853-1860, 1990) were moved from the original host into Campylobacter strains of a number of other Lior serogroups by a natural transformation procedure. This is the first report of the use of this transformation method to transfer a mutated locus among Campylobacter strains. Flagellin mutants were constructed in a number of heat-labile LIO serotypes and were serotyped and analyzed by immunoelectron microscopy with LIO typing antisera. In six cases, isogenic nonflagellated mutants were able to be serotyped in the same serogroup as their parent, and immunogold electron microscopy confirmed that antibodies in the typing antisera bound to components on the surface of both parent and mutant cells. However, in only one case, a strain belonging to serogroup LIO4, was a nonflagellated mutant untypeable, and immunogold electron microscopy showed that antibodies bound to the flagella filament of the parent but not to the cell surface. Furthermore, after introduction and expression as a flagellar filament of a LIO8 flagellin gene in this mutant, the strain could not be serotyped. These results indicate that a nonflagellar antigen is often the serodeterminant in the heat-labile Lior serotyping scheme.

Alleles↗
PubMed1991

Evaluation of 10 methods to distinguish epidemic-associated Campylobacter strains.

We compared four phenotypic and six genotypic methods for distinguishing Campylobacter jejuni strains from animals and humans involved in four epidemics. Based on a comparison with epidemiologic data, the methods that correctly identified all strains in three milkborne outbreaks and one waterborne outbreak were heat-stable and heat-labile serotyping; multilocus enzyme electrophoresis (MEE); DNA restriction endonuclease analysis with BglII, XhoI, PvuII, or PstI; and Southern blot and hybridization of PvuII- and PstI-digested DNA with Escherichia coli 16S and 23S rRNA (ribotyping). Biotyping, phage typing, plasmid analysis, and probing of BglII and XhoI DNA digests with C. jejuni 16S rRNA genes failed to correctly separate one or more strains. MEE, restriction endonuclease analysis, and ribotyping were the most sensitive methods and identified nine types among the 22 strains. These methods were also capable of further distinguishing strains within the same serotype. Data from MEE were also analyzed to calculate genetic relatedness among strains. Serotyping was the most discriminating phenotypic method, with eight and seven types distinguished by the heat-stable and heat-labile methods, respectively. MEE and ribotyping had several advantages over the other methods because they measure relatively stable and significant chromosomal differences and are applicable to other species and genera. These methods, however, are complex and not easily quantified; they are currently limited to specialized laboratories. When antisera are available, serotyping appears to be an effective and more practical approach to the identification of epidemic-related strains.

Animals↗
PubMed1991

Identification of verotoxin type 2 variant B subunit genes in Escherichia coli by the polymerase chain reaction and restriction fragment length polymorphism analysis.

A set of synthetic oligonucleotide primers was designed for use in a polymerase chain reaction protocol to specifically detect the B subunit genes in vtx2ha and vtx2hb, which code for the production of the VT2 (Shiga-like toxin II) variant cytotoxins VT2v-a and VT2v-b, respectively. An additional set of primers amplified a fragment common to the B subunits of the VT2 and the VT2 variant genes. Subsequent restriction endonuclease digestion of this amplicon permitted prediction of specific VT2 and variant genotypes on the basis of predetermined restriction fragment length polymorphisms. Genotypes of 21 VT2-producing strains of Escherichia coli were determined using this polymerase chain reaction-restriction fragment length polymorphism procedure. Four strains contained B subunit target sequences only for VT2 genes, 9 strains contained sequences only for VT2v-a genes, and 3 strains contained sequences only for VT2v-b. For genes in combination, one strain contained B subunit genes for both VT2 and VT2v-a and two strains contained B subunit genes for VT2 and VT2v-b. Two strains of E. coli O91:H21 contained both VT2v-a and VT2v-b B subunit genes. The VT2 reference strain of E. coli, E32511, was found to contain the targeted sequences from both VT2 and VT2v-a genes, whereas the recombinant E. coli, pEB1, possessed only that of the VT2 gene. The specific activities of extracellular VT2 determined in HeLa cells ranged from 0.3 to 41.7 TCD50 per microgram of protein in strains carrying the VT2 gene target and from 0 to 50.0 TCD50 per microgram of protein in strains carrying only the VT2 variant target (TCD50 is the tissue culture dose by which 50% of the cells were affected), suggesting that phenotypic expression does not correlate with genotype.

Animals↗
PubMed1991

Colicinogeny of O157:H7 enterohemorrhagic Escherichia coli and the shielding of colicin and phage receptors by their O-antigenic side chains.

Twenty O157:H7 enterohemorrhagic Escherichia coli strains from patients with different clinical conditions were tested for colicinogeny and the presence of Verotoxin (VT) genes. From bloody diarrhea cases, 7/8 isolates and from hemolytic uremic syndrome cases 3/5 isolates all synthesized what appeared to be colicin D. The remaining strains, which included 7 from asymptomatic sources, were noncolicinogenic. The plasmid determining the colicin was found to be 1.4 kb larger than the 5.2-kb pColD. The colicin D protein had a molecular weight of about 90,000, whereas the O157 colicin was 87,000. The plasmid was designated pColD157 to reflect these differences. Of O157:H7 isolates 17/20 had genes for both of the Verotoxins VT1 and VT2, and the remaining 3/20 for VT1 only. There was no correlation between the presence of VT determinants and colicinogeny or symptoms. The O157:H7 strains exhibited significant resistance to other colicins and bacteriophages.

Antigens, Bacterial↗
PubMed1990

Prevalence of verocytotoxigenic Escherichia coli in ground beef, pork, and chicken in southwestern Ontario.

Samples of ground beef (225), pork (235) and chicken (200) were randomly selected from meat processing plants in the southwestern Ontario area. Supernatants of broth cultures of the samples were tested for verocytotoxins using a Vero cell assay. Neutralization of cytotoxic activity using antisera specific for three types of verocytotoxin (Verotoxin 1, Verotoxin 2 and Shiga-like toxin II) was performed on positive samples. Isolation of verocytotoxigenic Escherichia coli (VTEC) was attempted from positive samples. VTEC were confirmed as E. coli biochemically, tested for drug resistance, and serotyped. Based on neutralization studies, the prevalence of VTEC in beef and pork was at least 36.4% and 10.6%, respectively. This is much higher than has been reported from a survey of retail meats in which a method designed to detect only E. coli O 157.H7 was used. Isolations of VTEC were made from 10.4% of the beef samples and 3.8% of the pork samples. No VTEC were recovered from the chicken samples. The majority of VTEC isolates were susceptible to commonly used antimicrobial agents. A number of the serotypes of the VTEC isolates recovered have been associated with human disease; however, no VTEC of serotype O 157.H7 were isolated.

Animals↗
PubMed1990

Extended phage-typing scheme for Escherichia coli O157:H7.

In Canada, the number of human isolates of verotoxigenic (VT + ve) Escherichia coli O157:H7 from diarrhoeal cases and haemolytic uraemic syndrome and haemorrhagic colitis has increased from 25 in 1982 to 2384 in 1989. A total of 3273 VT + ve E. coli O157:H7 strains (3255 strains isolated in Canada and 18 isolates from other countries) were phage typed. The phage typing scheme has been extended from 14 to 62 phage types. Of these, five types occurred exclusively in other countries (type 47 in Japan; and types 49, 50, 51 and 52 in the U.K.). Thirty-five different phage types were identified in Canada; only nine of these (1, 2, 4, 8, 14, 21, 23, 31 and 32), each accounted for more than 1% of the cases from human sources. The same nine types were the only ones observed among the isolates from non-human sources (meat and slaughter houses) suggesting a food-borne transmission in most of the human cases. Phage types 1 (30.5%); 4 (21%); 8 (13.5%); 31 (8.9%) and 14 (8%) were encountered in varying frequencies in most of the provinces; infrequently occurring phage types also showed regional variation. Thirteen different phage types were identified among 151 outbreaks representing 556 isolates of E. coli O157:H7. More than one phage type were encountered in 12 outbreaks whereas in 141 outbreaks, all strains in each, had the same phage type.

Animals↗
PubMed1990

Differentiation of Shiga toxin and Vero cytotoxin type 1 genes by polymerase chain reaction.

Two sets of synthetic oligonucleotide primers were used in a polymerase chain reaction technique to distinguish genes for Shiga toxin in Shigella dysenteriae 1 and type 1 Vero cytotoxin (VT1) in Escherichia coli. VT1a and VT1b primers directed at a common 130-base-pair (bp) fragment of the stx and sltI genes detected template nucleic acid in both Shiga toxin-positive S. dysenteriae 1 and VT1-producing E. coli strains. VT1c and VT1d primers, targeting a 140-bp fragment of the promoter region of the sltIA gene, were negative in the polymerase chain reaction with S. dysenteriae 1 nucleic acid and positive with nucleic acids from all strains found to produce VT1 in toxin-specific neutralization tests. Primer specificity was determined in the polymerase chain reaction using nucleic acid extracted from 49 strains of representative enteric pathogens defined in terms of their toxigenicity.

Bacterial Toxins↗
PubMed1990

Detection of the aerolysin gene in Aeromonas hydrophila by the polymerase chain reaction.

Synthetic oligonucleotide primers were used in a polymerase chain reaction (PCR) technique to detect the gene for aerolysin in strains of Aeromonas hydrophila and to screen for identical genes in A. caviae, A. sobria, and A. veronii isolated from patients with diarrheal disease. Primers targeted a 209-bp fragment of the aer gene coding for the beta-hemolysin and detected template DNA only in the PCR using nucleic acid (NA) from hemolytic strains of A. hydrophila which were also cytotoxic to Vero and CHO cells and enterotoxic in suckling-mouse assays. PCR amplification of NA from hemolytic A. sobria or nonhemolytic A. hydrophila and A. caviae strains was consistently negative. Primer specificity was determined in the PCR by using NA extracted from 56 strains of bacteria, including hemolytic Escherichia coli and Listeria monocytogenes as well as several recognized enteric pathogens defined in terms of their toxigenicity. The detection limit for the aerolysin gene by PCR amplification was 1 ng of total NA. The PCR clearly identified aerolysin-producing strains of A. hydrophila and may have application as a species-specific virulence test because other hemolytic Aeromonas species tested were negative.

Aeromonas↗
PubMed1990

Rapid and specific detection of verotoxin genes in Escherichia coli by the polymerase chain reaction.

A set of four synthetic oligonucleotide probes derived from sequences of the VT1 (Shiga-like toxin I [SLT-I]) and VT2 (SLT-II) genes were used in a polymerase chain reaction (PCR) amplification procedure to detect these genes in some enteric pathogens. A total of 40 verotoxin-producing Escherichia coli strains and 43 isolates of other recognized enteric pathogens were studied. PCR amplification products identifying the VT1 and VT2 gene sequences were observed only in nucleic acid extracted from strains found to be VT positive in traditional tissue culture assays. Template nucleic acid extracted from other gram-negative bacteria was found to be negative with the exception of five isolates of Shigella dysenteriae type 1 in which good amplification with the VT1 probe was observed. The oligonucleotide probes clearly distinguished VT1 and VT2 strains of E. coli and did not give specific amplification with nucleic acid from VTe (a SLT-II variant)-producing E. coli. VT1 or VT2 genes or both were not detected in E. coli K-12 strain C600 or HB101 or in strains known to express other virulence factors, such as enterotoxins, adhesins, hemolysins, or unrelated cytotoxins. The sensitivity of the PCR procedure for detection of both VT1 and VT2 genes was determined to be 1 ng of total nucleic acid. Furthermore, the VT1 gene was easily detected when only 100 pg of nucleic acid was used as the template in the PCR procedure.

Animals↗
PubMed1990

Virulence-related genes in ColV plasmids of Escherichia coli isolated from human blood and intestines.

DNA probes for the colicin V, traT, iss, and iu genes were used in this study of four representative ColV plasmids together with 200 Escherichia coli strains isolated from the stools of patients with diarrhea and 146 E. coli strains isolated from the blood of patients with bacteremia. The study indicated that the ColV plasmids are heterogeneous. Southern and colony hybridization analyses showed that in most of the colicin V-producing intestinal E. coli strains, the colicin V genes are located in the chromosome (14 of 16); in most of the colicin V-producing E. coli strains isolated from the blood, they are located in plasmids (18 of 22). In both intestinal and blood E. coli isolates, the traT, iss, and aerobactin receptor genes were present at similar frequencies, but the frequency of the aerobactin synthesis genes was significantly different. The aerobactin receptor gene was present in 25% of the intestinal E. coli strains that lack the aerobactin synthesis gene. In the blood isolates, the aerobactin synthesis and receptor genes were present at almost equal frequencies. Among the colicin V-producing isolates, the iss, traT, and iu genes were present in 95.5, 86.4, and 90.9% of the blood isolates and in only 68.8, 43.8, and 81.3% of the intestinal isolates, respectively. The ColV plasmids from blood isolates that were tested for the presence of traT, iss, and iu genes were homogeneous and had DNA sequences that hybridized with each of the probes. On the other hand, the two intestinal strains containing ColV genes in a plasmid were heterogeneous in regard to the carriage of these genes. The presence of ColV is not restricted to specific O types.

Blood↗