Typing of sequential bacterial isolates by pulsed-field gel electrophoresis

doi:10.1016/0732-8893(95)00139-8

Diagnostic Microbiology and Infectious Disease

Volume 22, Issue 4, August 1995, Pages 309-314

https://doi.org/10.1016/0732-8893(95)00139-8 Get rights and content

Abstract

We typed 39 sets of multiple bacterial isolates of the same species from patients by pulsed-field gel electrophoresis of genomic DNA (PFGE). Isolates were cultured from different sites or over a 2-week or longer interval. Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae were tested. Excluding E. cloacae, 28 of 32 sets of isolates (87%) demonstrated only identical or highly related PFGE types. Four of the seven sets of E. cloacae showed different types. For species other than E. cloacae, our results suggest that patients are usually colonized and infected with a single strain of these bacterial pathogens. Unlike all of the other tested species, E. cloacae PFGE typing differences suggested the presence of multiple strains causing colonization and infection.

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Outbreak of Neisseria meningitidis C in a Brazilian oil refinery involving an adjacent community
2013, Enfermedades Infecciosas y Microbiologia Clinica
Citation Excerpt :
The isolates showing similarity >85% were considered genetically related, i.e. belonging to the same clone. The isolates considered to be related were tabulated by using the same capital letter and numbers if the isolates were non-identical but closely related.18,19 Serotyping for all isolates was performed by dot-blotting using whole cell suspensions as previously described.20
An outbreak of serogroup C meningococcal disease (MD) that involved employees from an oil refinery occurred in Paulínia, from March to June 2010, and spread to the community of Cosmópolis, both situated in São Paulo State, Brazil. The aim of this study was to describe the epidemiological and laboratory aspects, and the control measures that were implemented.
Descriptive and molecular epidemiological analysis was used to define the extent of the outbreak and the common risk factors among outbreak related cases. Vaccination initiative targeted the employees from the oil refinery and the community-based individuals.
A total of six outbreak-related cases of MD in the oil refinery and 12 cases in the city of Cosmópolis, São Paulo State occurred in a three-month period. All 18 MD cases were confirmed as serogroup C either by culture, real time polymerase chain reaction (RT-PCR), or counterimmunoelectrophoresis (CIE). The meningococcal isolates (n = 15) had the phenotype C:23:P1.14-6 and showed close genetic relationship by pulsed field gel electrophoresis (PFGE). Multilocus sequence typing (MLST) characterization showed that the clones were ST 3780, ST-103 complex. The isolates were susceptible to the antibiotics tested. Vaccination was administered to 15,848 persons at the oil refinery and 18,571 persons of the community.
The molecular characterization of the Neisseria meningitidis (N. meningitidis) strains, and the efficient investigation and the prompt measures implemented were essential for controlling the disease in the region.
De marzo a junio de 2010 se produjo un brote de enfermedad meningocócica (EM) del serogrupo C en una refinería de petróleo en la ciudad de Paulina, afectando a los empleados y a la comunidad de Cosmópolis, ambas ciudades situadas en el estado de Sao Paulo (Brasil). Los objetivos del estudio fueron describir los aspectos epidemiológicos y de laboratorio así como las medidas de control que se implementaron.
Se realizo un análisis epidemiológico descriptivo y molecular para definir la extensión de la epidemia y los factores de riesgo comunes entre los casos relacionados con el brote. Se utilizo vacuna para los empleados de la refinería de petróleo y sus contactos en la comunidad.
Un total de seis casos fueron relacionados con el brote de EM en la refinería de petróleo y 12 casos en la ciudad de Cosmópolis, durante un período de tres meses. Dieciocho casos de EM fueron confirmados como del serogrupo C mediante cultivo, reacción en cadena de la polimerasa en tiempo real (RT-PCR) o contrainmunoelectroforesis (CIE). Las cepas de meningococo aisladas (n = 15) presentaban el fenotipo C: 23: P1.14-6 y mostraron estrecha relación genética por PFGE. La caracterización de MLST demostró que las cepas eran ST 3780, complejo ST-103 y eran susceptibles a los antibióticos probados. La vacunación se administró a 15.848 personas en la refinería y a 18.571 personas de la comunidad.
La caracterización molecular de las cepas de N. meningitidis, la investigación eficaz y rápida y las medidas implementadas fueron esenciales para controlar la enfermedad en la región.
Molecular characterization of Neisseria meningitidis B: NT: P1.14/162 clonal complex responsible of invasive meningococcal disease in the north of Italy
2012, Diagnostic Microbiology and Infectious Disease
Study of an outbreak of Enterobacter cloacae sepsis in a neonatal intensive care unit: The application of epidemiologic chromosome profiling by pulsed-field gel electrophoresis
2002, American Journal of Infection Control
Citation Excerpt :
This finding is consistent with the results reported by Gaston,1 who found that an increased isolation rate of E cloacae in a particular hospital ward was a result of the presence of multiple strains rather than a single strain. This finding has also been confirmed in other studies,8-10 but such reports are rare. In the present study, strain I and II were the most frequently isolated strains during the outbreak.
Objective: From October 1996 to March 1997, a cluster of 11 cases of neonatal sepsis caused by Enterobacter cloacae with similar antimicrobial susceptibility patterns occurred in a neonatal intensive care unit. This outbreak prompted an investigation. Method: Twelve isolates obtained from 6 neonatal patients who developed E cloacae sepsis during the outbreak were analyzed. Four E cloacae isolates from 2 preterm neonates without E cloacae infection on the same ward, and 1 isolate from the hands of a nurse, were also examined. No E cloacae were isolated from the environment. Bacterial DNA digested with Xba I or Not I was analyzed with pulsed-field gel electrophoresis. Result: Three distinct banding patterns were identified by pulsed-field gel electrophoresis. Of the 6 preterm infants with sepsis, strain I was identified in 1, strain II in 2, a mixed infection of strains I and II in 2, and strain III was found in only 1 infant. An isolate from the hands of a nurse was identified as strain II , as were the 4 isolates from the 2 preterm neonates without E cloacae infection. Thus, this outbreak of sepsis was caused by 2 genotypes of E cloacae. Conclusion: This study demonstrates that pulsed-field gel electrophoresis with restriction enzyme digestion is a valuable tool for genetic characterization of multidrug-resistant E cloacae strains during outbreaks. (Am J Infect Control 2002;30:381-5)
Outbreak of Neisseria meningitidis C in workers at a large food-processing plant in Brazil: Challenges of controlling disease spread to the larger community
2012, Epidemiology and Infection
Neisseria meningitidis Serogroup X sequence type 2888, Italy
2010, Emerging Infectious Diseases
Serogroup C meningococci in Italy in the era of conjugate menC vaccination
2009, BMC Infectious Diseases

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^☆: This work was presented in part at the 31st Annual Meeting of the Infectious Diseases Society of America , 16–18 October 1993, New Orleans, LA (Abstract no. 150).

¹: Dr. Chetchotisakd is at Khonkaen University, P.O. Box 200, Khonkaen 4002, Thailand.

²: Mr. Phelps is at Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN 46285, USA.

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Diagnostic Microbiology and Infectious Disease

Abstract

References (17)

Purification, specific fragmentation and separation of large DNA molecules

Methods Enzymol

Resolution of recent evolutionary divergence among Escherichia coli from related lineages: the application of pulsed field electrophoresis to molecular epidemiology

J Infect Dis

Assessment of bacterial cross-transmission as a cause of infections in patients in intensive care units

Clin Infect Dis

Measures of the amount of ecologic association between species

Ecology

Comparison of ribotyping with conventional methods for the type identification of Enterobacter cloacae

J Clin Microbiol

Molecular epidemiology of nosocomial infection: analysis of chromosomal restriction fragment patterns by pulsed-field gel electrophoresis

Infect Control Hosp Epidemiol

Typing of Enterococcus species by DNA restriction fragment analysis

J Clin Microbiol

Are point mutations or DNA rearrangements responsible for the restriction fragment length polymorphisms that are used to type bacteria?

Microbiology

Cited by (13)

Outbreak of Neisseria meningitidis C in a Brazilian oil refinery involving an adjacent community

Molecular characterization of Neisseria meningitidis B: NT: P1.14/162 clonal complex responsible of invasive meningococcal disease in the north of Italy

Study of an outbreak of Enterobacter cloacae sepsis in a neonatal intensive care unit: The application of epidemiologic chromosome profiling by pulsed-field gel electrophoresis

Outbreak of Neisseria meningitidis C in workers at a large food-processing plant in Brazil: Challenges of controlling disease spread to the larger community

Neisseria meningitidis Serogroup X sequence type 2888, Italy

Serogroup C meningococci in Italy in the era of conjugate menC vaccination

Typing of sequential bacterial isolates by pulsed-field gel electrophoresis☆

Abstract

Methods Enzymol

Resolution of recent evolutionary divergence among Escherichia coli from related lineages: the application of pulsed field electrophoresis to molecular epidemiology

J Infect Dis

Assessment of bacterial cross-transmission as a cause of infections in patients in intensive care units

Clin Infect Dis

Measures of the amount of ecologic association between species

Ecology

Comparison of ribotyping with conventional methods for the type identification of Enterobacter cloacae

J Clin Microbiol

Molecular epidemiology of nosocomial infection: analysis of chromosomal restriction fragment patterns by pulsed-field gel electrophoresis

Infect Control Hosp Epidemiol

Typing of Enterococcus species by DNA restriction fragment analysis

J Clin Microbiol

Are point mutations or DNA rearrangements responsible for the restriction fragment length polymorphisms that are used to type bacteria?

Microbiology