Pulsed-Field Gel Electrophoresis Used To Investigate Genetic Diversity of Haemophilus influenzae Type b Isolates in Australia Shows Differences between Aboriginal and Non-Aboriginal Isolates

1999 ◽  
Vol 37 (5) ◽  
pp. 1524-1531 ◽  
Author(s):  
Patricia Ezekiel Moor ◽  
Peter C. Collignon ◽  
Gwendolyn L. Gilbert
Keyword(s):  
Population Structure ◽  
Haemophilus Influenzae ◽  
Gel Electrophoresis ◽  
Strong Association ◽  
Pulsed Field Gel Electrophoresis ◽  
Haemophilus Influenzae Type ◽  
Type B ◽  
Pulsed Field ◽  
Rflp Type ◽  
Aboriginal Children

We used pulsed-field gel electrophoresis to study the epidemiology and population structure of Haemophilus influenzae type b. DNAs from 187 isolates recovered between 1985 and 1993 from Aboriginal children (n = 76), non-Aboriginal children (n = 106), and non-Aboriginal adults (n = 5) in urban and rural regions across Australia were digested with the SmaI restriction endonuclease. Patterns of 13 to 17 well-resolved fragments (size range, ∼8 to 500 kb) defining 67 restriction fragment length polymorphism (RFLP) types were found. Two types predominated. One type (n = 37) accounted for 35 (46%) of the isolates from Aboriginals and 2 (2%) of the isolates from non-Aboriginals, and the other type (n = 41) accounted for 2 (3%) of the isolates from Aboriginals and 39 (35%) of the isolates from non-Aboriginals. Clustering revealed seven groups at a genetic distance of ∼50% similarity in a tree-like dendrogram. They included two highly divergent groups representing 50 (66%) isolates from Aboriginals and 6 (5%) isolates from non-Aboriginals and another genetically distinct group representing 7 (9%) isolates from Aboriginals and 81 (73%) isolates from non-Aboriginals. The results showed a heterogeneous clonal population structure, with the isolates of two types accounting for 42% of the sample. There was no association between RFLP type and the diagnosis of meningitis or epiglottitis, age, sex, date of collection, or geographic location, but there was a strong association between the origin of isolates from Aboriginal children and RFLP type F2a and the origin of isolates from non-Aboriginal children and RFLP type A8b. The methodology discriminated well among the isolates (D = 0.91) and will be useful for the monitoring of postvaccine isolates of H. influenzae type b.

scholarly journals Characterization of Non-Type B Haemophilus influenzae Strains Isolated from Patients with Invasive Disease

2000 ◽  
Vol 38 (12) ◽  
pp. 4649-4652 ◽  
Author(s):  
Marina Cerquetti ◽  
Marta Luisa Ciofi degli Atti ◽  
Giovanna Renna ◽  
Alberto Eugenio Tozzi ◽  
Maria Laura Garlaschi ◽  
...  
Keyword(s):  
Haemophilus Influenzae ◽  
Gel Electrophoresis ◽  
Invasive Disease ◽  
Pulsed Field Gel Electrophoresis ◽  
Type B ◽  
Pulsed Field ◽  
Clonal Spread

Forty-one non-type b Haemophilus influenzae isolates from cases of invasive disease were characterized. By PCR capsular genotyping, 33 nonencapsulated strains, 4 type f isolates, and 4 b− strains were identified. By pulsed-field gel electrophoresis, the nonencapsulated isolates exhibited great genetic heterogenicity, whereas the type f and the b− strains seemed to have a clonal spread. Occurrence of the hifA gene was found by PCR in 18% of the nonencapsulated, 50% of the b−, and all of the type f strains. Hemagglutinating fimbriae were generally expressed by nonencapsulated isolates when fimbrial gene hifA was present. Two nonencapsulated isolates not susceptible to ampicillin were detected; no strains were positive for β-lactamase production.

scholarly journals Association of IS1016 with the hia Adhesin Gene and Biotypes V and I in Invasive Nontypeable Haemophilus influenzae

2008 ◽  
Vol 76 (11) ◽  
pp. 5221-5227 ◽  
Author(s):  
Sarah W. Satola ◽  
Brooke Napier ◽  
Monica M. Farley
Keyword(s):  
Genetic Diversity ◽  
Haemophilus Influenzae ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Division I ◽  
Emerging Infections ◽  
Nontypeable Haemophilus Influenzae ◽  
Insertion Element ◽  
Pulsed Field ◽  
Slide Agglutination

ABSTRACT A subset of invasive nontypeable Haemophilus influenzae (NTHI) strains has evidence of IS1016, an insertion element associated with division I H. influenzae capsule serotypes. We examined IS1016-positive invasive NTHI isolates collected as part of Active Bacterial Core Surveillance within the Georgia Emerging Infections Program for the presence or absence of hmw1 and hmw2 (two related adhesin genes that are common in NTHI but absent in encapsulated H. influenzae) and hia (homologue of hsf, an encapsulated H. influenzae adhesin gene). Isolates were serotyped using slide agglutination, confirmed as NTHI strains using PCR capsule typing, and biotyped. Two hundred twenty-nine invasive NTHI isolates collected between August 1998 and December 2006 were screened for IS1016; 22/229 (9.6%) were positive. Nineteen of 201 previously identified IS1016-positive invasive NTHI isolates collected between January 1989 and July 1998 were also examined. Forty-one IS1016-positive and 56 randomly selected IS1016-negative invasive NTHI strains were examined. The hia adhesin was present in 39 of 41 (95%) IS1016-positive NTHI strains and 1 of 56 (1.8%) IS1016-negative NTHI strains tested; hmw (hmw1, hmw2, or both) was present in 50 of 56 (89%) IS1016-negative NTHI isolates but in only 5 of 41 (12%; all hmw2) IS1016-positive NTHI isolates. IS1016-positive NTHI strains were more often biotype V (P < 0.001) or biotype I (P = 0.04) than IS1016-negative NTHI strains, which were most often biotype II. Pulsed-field gel electrophoresis revealed the expected genetic diversity of NTHI with some clustering based on IS1016, hmw or hia, and biotypes. A significant association of IS1016 with biotypes V and I and the presence of hia adhesins was found among invasive NTHI. IS1016-positive NTHI strains may represent a unique subset of NTHI strains, with characteristics more closely resembling those of encapsulated H. influenzae.

scholarly journals Subtyping of Haemophilus influenzaeStrains by Pulsed-Field Gel Electrophoresis

1999 ◽  
Vol 37 (7) ◽  
pp. 2142-2147 ◽  
Author(s):  
Mitsumasa Saito ◽  
Akiko Umeda ◽  
Shin-ichi Yoshida
Keyword(s):  
Cerebrospinal Fluid ◽  
Haemophilus Influenzae ◽  
Gel Electrophoresis ◽  
Invasive Disease ◽  
Pulsed Field Gel Electrophoresis ◽  
Pfge Analysis ◽  
Pulsed Field ◽  
Pfge Typing ◽  
Household Contacts ◽  
Serotype B

A total of 200 isolates of Haemophilus influenzae were analyzed by serotyping, biotyping, and pulsed-field gel electrophoresis (PFGE). A total of 178 epidemiologically unrelated strains of H. influenzae demonstrated a variety of genome patterns by PFGE, and 165 genotypes were thus obtained in this study. PFGE typing proved to have a much stronger discriminatory power than either serotyping or biotyping. Six serotype b strains were all classified into discrete genotypes. A PFGE analysis of 18 strains obtained from the nasopharynx, blood, and cerebrospinal fluid of patients with meningitis also supported the hypothesis that invasive H. influenzaedisseminates from the nasopharynx to the bloodstream and then subsequently to other body sites. PFGE typing of 10 other strains isolated from household contacts of patients with H. influenzae infection revealed that the strain that caused theH. influenzae infection often colonized the nasopharynges of household contacts. Our findings suggest that PFGE analysis is useful for the epidemiological study of H. influenzaeinfection, even when the invasive disease is caused by serotype b strains.

scholarly journals INVASIVE NON-TYPE B HAEMOPHILUS INFLUENZAE DISEASE: REPORT OF EIGHT CASES

2019 ◽  
Vol 37 (2) ◽  
pp. 156-160 ◽  
Author(s):  
Mariana Tresoldi das Neves Romaneli ◽  
Antonia Teresinha Tresoldi ◽  
Ricardo Mendes Pereira ◽  
Márcia Teixeira Garcia ◽  
Christian Cruz Hofling ◽  
...  
Keyword(s):  
Haemophilus Influenzae ◽  
Molecular Typing ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Nosocomial Transmission ◽  
Reference Laboratory ◽  
Beta Lactamase ◽  
Type B ◽  
Type C ◽  
Diagnose Monitor

ASTRACT Objective: To describe eight cases of invasive non-type b Haemophilus influenzae disease in children admitted to Hospital de Clínicas of Universidade Estadual de Campinas. Cases description: In 2015, there were eight cases of invasive non-type b H. influenzae disease. We tested the ampicillin sensitivity and beta-lactamase production of the strains identified and performed the genotyping. Molecular typing was determined by Pulsed-Field Gel Electrophoresis. Four patients were diagnosed with bacteremia; in two cases, H. influenzae was detected in the pleural fluid, and two patients had meningitis. Patients with comorbidities represented 37.5% of cases. Except for the strain of one patient - not sent to the reference laboratory -, all were ampicillin-sensitive and non-beta-lactamase-producing. Genotyping identified four non-capsular, one type c, and two type a strains. Molecular typing ruled out nosocomial transmission since all serotypes were distinct regarding genotype. Comments: The rise in cases of invasive non-type b H. influenzae infection was real. There was no nosocomial transmission, and we found no justification for the increase. These data indicate the need for surveillance to correctly diagnose, monitor, and understand the spectrum of non-type b H. influenzae disease.

scholarly journals The effectiveness of Haemophilus influenzae type b conjugate vaccines in a high risk population measured using immunization register data

2001 ◽  
Vol 126 (1) ◽  
pp. 31-36 ◽  
Author(s):  
P. MARKEY ◽  
V. KRAUSE ◽  
J. W. BOSLEGO ◽  
P. M. COPLAN ◽  
J.M. DARGAN ◽  
...  
Keyword(s):  
High Risk ◽  
Haemophilus Influenzae ◽  
Incidence Rates ◽  
Haemophilus Influenzae Type ◽  
High Risk Population ◽  
Type B ◽  
Vaccine Introduction ◽  
Immunization Rates ◽  
Aboriginal Children ◽  
The Impact

The Northern Territory of Australia has had historically very high incidence rates of invasive Haemophilus influenzae type b disease in children less than 5 years of age, with the burden of disease greatest among Aboriginal infants less than 12 months. This study documents the impact of conjugate Hib vaccines introduced in 1993. Immunization rates were monitored using an existing immunization register, and case finding was done retrospectively using hospital and laboratory records. Following the vaccine introduction, the incidence fell abruptly to a seventh of its pre-vaccination level, in both Aboriginal and non-Aboriginal children. The effectiveness of PRP-OMPC (PedvaxHIBTM) was 97·5% and the overall effectiveness of the vaccination programme was 86·3%. The study shows Hib immunization as an effective intervention while discussing continuing needs for Hib control in high risk populations. It also illustrates the benefit of immunization registers in the evaluation of immunization programmes and assessment of vaccine effectiveness.

scholarly journals Streptococcus bovis Clone Causing Two Episodes of Endocarditis 8 Years Apart

1999 ◽  
Vol 37 (3) ◽  
pp. 862-863 ◽  
Author(s):  
Kathrin Mühlemann ◽  
Susanne Graf ◽  
Martin G. Täuber
Keyword(s):  
Population Structure ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Heterogeneous Population ◽  
Streptococcus Bovis ◽  
Pulsed Field

A patient had endocarditis caused by Streptococcus bovis twice 8 years apart. According to pulsed-field gel electrophoresis (PFGE) the two isolates were identical. Seven unrelated blood isolates of S. bovis yielded unique PFGE patterns. Considering this heterogeneous population structure our findings demonstrate the long-term persistence of an S. bovis clone in a patient with recurrent endocarditis.

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