Evaluation of DNA Fingerprinting by PFGE as an Epidemiologic Tool for <i>Salmonella</i> Infections

Murase, Toshiyuki; Okitsu, Tadayuki; Suzuki, Riéko; Morozumi, Hirotoshi; Matsushima, Akiyoshi; Nakamura, Akiko; Yamai, Shiro

doi:10.1111/j.1348-0421.1995.tb03255.x

Cited by 64 publications

(42 citation statements)

References 12 publications

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“…Some genotyping methods, such as RFLP of 16s rRNA and IS200 and low-frequency restriction analysis of chromosomal DNA with PFGE have proved to be helpful for the epidemiological typing of Typhimurium [56]. PFGE may provide evidence of minor genetic heterogeneity which cannot be detected readily by such RFLP-based techniques as ribotyping and IS200 fingerprinting.…”

Section: Discussionmentioning

confidence: 99%

Increase in incidence of resistance to ampicillin, chloramphenicol and trimethoprim in clinical isolates of Salmonella serotype Typhimurium with investigation of molecular epidemiology and mechanisms of resistance

Gallardo¹,

Ruiz²,

Marco³

et al. 1999

Journal of Medical Microbiology

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Antimicrobial resistance patterns of Salmonella serotype Typhimurium isolates obtained during the period 1987-1994 were examined and the molecular epidemiology and the mechanisms of resistance to ampicillin, chloramphenicol and trimethoprim were investigated in 24 strains isolated during 1994. Resistance to ampicillin increased from 18% to 78%, to chloramphenicol from 15y0 to 78%, to tetracycline from 53% to 89% and to co-trimoxazole from 3% to 37%, whereas resistance to norfloxacin remained at 0%. Of Salmonella serotype Typhimurium strains isolated during 1994, all ampicillinresistant strains had an MIC > 256 mg/L, except one strain in which the MIC was 64 mg/L. Twelve strains (52%) had a TEM-type /3-lactamase, nine (39%) a CARB-type /3-lactamase and two strains (8%) had an OXA-type /3-lactamase. Chloramphenicol acetyl-transferase activity was detected in only nine (47%) of 19 chloramphenicol resistant strains, whereas all eight trimethoprim-resistant strains produced a dihydrofolate reductase type Ia enzyme. Three different epidemiological groups were defined by either low-frequency restriction analysis of chromosomal DNA and pulsed-field gel electrophoresis or repetitive extragenic palindromic-PCR. The latter technique provided an alternative, rapid and powerful genotyping method for S. Typhimurium. Although quinolones provide a good therapeutic alternative, the multiresistance of S. Typhimurium is of public health concern and it is important to continue surveillance of resistance levels and their mechanisms.

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Section: Discussionmentioning

confidence: 99%

Increase in incidence of resistance to ampicillin, chloramphenicol and trimethoprim in clinical isolates of Salmonella serotype Typhimurium with investigation of molecular epidemiology and mechanisms of resistance

Gallardo¹,

Ruiz²,

Marco³

et al. 1999

Journal of Medical Microbiology

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“…Pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility typing (AST) are two commonly used methods for studying microbial epidemiology and trends in the antibiotic resistance of bacteria. PFGE is currently used by the CDC PulseNet surveillance program and is generally accepted as the "gold standard" for molecular typing of Salmonella (7,18,34,41,57). Due to its limitations, many studies have compared PFGE to other genetic typing methods in attempts to identify more powerful tools for epidemiological investigations and evolutionary analyses.…”

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confidence: 99%

Comparison of Multilocus Sequence Typing, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Typing for Characterization of Salmonella enterica Serotype Newport Isolates

et al. 2006

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In the United States, multidrug-resistant phenotypes of Salmonella enterica serotype Newport (commonly referred to as MDR-AmpC) have emerged in animals and humans and have become a major public health problem. Although pulsed-field gel electrophoresis (PFGE) is the current "gold standard" typing method for Salmonella, multilocus sequence typing (MLST) may be more relevant to investigations exploring evolutionary and population biology relationships. In this study, 81 Salmonella enterica serotype Newport isolates from humans, food animals, and retail foods were examined for antimicrobial susceptibility and characterized using PFGE and MLST of seven genes, aroC, dnaN, hemD, hisD, purE, sucA, and thrA. Forty-nine percent of the isolates were resistant to nine or more of the tested antimicrobials. Salmonella isolates displayed resistance most often to sulfamethoxazole (57%), streptomycin (56%), tetracycline (56%), ampicillin (52%), and ceftiofur (49%) and, to a lesser extent, to kanamycin (19%), trimethoprim-sulfamethoxazole (17%), and gentamicin (11%). A total of 43 PFGE patterns were generated using XbaI, indicating a genetically diverse population. The largest PFGE cluster contained isolates from clinically ill swine, cattle, and humans. MLST resulted in 12 sequence types (STs), with one type encompassing 62% of the strains. Ten new sequence types and one novel allele type were identified. Furthermore, MLST typing showed that strains closely related by PFGE clustered in major STs, whereas more distantly related strains were separated into two clusters by PFGE. The results of this study demonstrated that the MLST scheme employed here clustered S. enterica serovar Newport isolates in distinct molecular populations, and strain discrimination was enhanced by combining PFGE, antimicrobial susceptibility, and MLST results.

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“…Pulsed-field gel electrophoresis (PFGE) is an established method for the analysis of large fragments generated by restriction endonuclease digestion of genomic DNA (14) and is currently considered to be one of the most reliable typing procedures (16). The PFGE method has been shown to be highly effective for epidemiological studies of some serovars of S. enterica (2,3,17,20,21).…”

mentioning

confidence: 99%

Suitability of PCR Fingerprinting, Infrequent-Restriction-Site PCR, and Pulsed-Field Gel Electrophoresis, Combined with Computerized Gel Analysis, in Library Typing of Salmonella enterica Serovar Enteritidis

Garaizar

López-Molina

Laconcha

et al. 2000

Appl Environ Microbiol

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Strains of Salmonella enterica (n ‫؍‬ 212) of different serovars and phage types were used to establish a library typing computerized system for serovar Enteritidis on the basis of PCR fingerprinting, infrequent-restrictionsite PCR (IRS-PCR), or pulsed-field gel electrophoresis (PFGE). The rate of PCR fingerprinting interassay and intercenter reproducibility was low and was only increased when DNA samples were extracted at the same time and amplified with the same reaction mixtures. Reproducibility of IRS-PCR technique reached 100%, but discrimination was low (D ‫؍‬ 0.52). The PFGE procedure showed an intercenter reproducibility value of 93.3%. The high reproducibility of PFGE combined with the previously determined high discrimination directed its use for library typing. The use of PFGE with enzymes XbaI, BlnI, and SpeI for library typing of serovar Enteritidis was assessed with GelCompar 4.0 software. Three computer libraries of PFGE DNA profiles were constructed, and their ability to recognize new DNA profiles was analyzed. The results obtained pointed out that the combination of PFGE with computerized analysis could be suitable in long-term epidemiological comparison and surveillance of Salmonella serovar Enteritidis, specially if the prevalence of genetic events that could be responsible for changes in PFGE profiles in this serovar was low.

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Evaluation of DNA Fingerprinting by PFGE as an Epidemiologic Tool for Salmonella Infections

Cited by 64 publications

References 12 publications

Increase in incidence of resistance to ampicillin, chloramphenicol and trimethoprim in clinical isolates of Salmonella serotype Typhimurium with investigation of molecular epidemiology and mechanisms of resistance

Increase in incidence of resistance to ampicillin, chloramphenicol and trimethoprim in clinical isolates of Salmonella serotype Typhimurium with investigation of molecular epidemiology and mechanisms of resistance

Comparison of Multilocus Sequence Typing, Pulsed-Field Gel Electrophoresis, and Antimicrobial Susceptibility Typing for Characterization of Salmonella enterica Serotype Newport Isolates

Suitability of PCR Fingerprinting, Infrequent-Restriction-Site PCR, and Pulsed-Field Gel Electrophoresis, Combined with Computerized Gel Analysis, in Library Typing of Salmonella enterica Serovar Enteritidis

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