Improved pulsed field gel electrophoresis method for Moraxella catarrhalis

2008 ◽  
Vol 75 (2) ◽  
pp. 344-345 ◽  
Author(s):  
Nevada M. Pingault ◽  
Deborah Lehmann ◽  
Thomas V. Riley
1999 ◽  
Vol 45 (4) ◽  
pp. 299-303 ◽  
Author(s):  
K T Nguyen ◽  
E J Hansen ◽  
M A Farinha

A physical genome map of the Moraxella catarrhalis type strain (ATCC 25238) has been constructed using pulsed field gel electrophoresis. Macrorestriction analyses of the genome of M. catarrhalis were performed by digestion with the restriction enzymes SmaI, NotI, and RsrII, which cleave the single circular chromosome into 9, 10, and 6 fragments, respectively. The chromosomal fragments generated by pulsed field gel electrophoresis were converted to a linkage map utilizing a combination of partial digestions, and cross-hybridizations. Moraxella catarrhalis, like a number of other respiratory pathogens, has a relatively small genome estimated at 1750 kilobase pairs or about 40% of the size of the Escherichia coli genome. The locations of the four ribosomal RNA operons (rrnLS) were determined by Southern hybridization and by digestion with I-CeuI endonuclease. A number of genes involved in virulence have been placed onto the physical map by Southern hybridization including those encoding the predominant outer-membrane proteins and the chromosomal gene encoding beta-lactamase.Key words: Moraxella catarrhalis, physical map, genome analysis, pulsed-field gel electrophoresis, virulence.


2004 ◽  
Vol 70 (4) ◽  
pp. 1964-1972 ◽  
Author(s):  
Chien-Hsien Chen ◽  
Toshio Shimada ◽  
Nasreldin Elhadi ◽  
Son Radu ◽  
Mitsuaki Nishibuchi

ABSTRACT Of 97 strains of Vibrio cholerae isolated from various seafoods in Malaysia in 1998 and 1999, 20 strains carried the ctx gene and produced cholera toxin. Fourteen, one, and five of these toxigenic strains belonged to the O139, O1 Ogawa, and rough serotypes, respectively. The rough strains had the rfb gene of the O1 serotype. The toxigenic strains varied in their biochemical characteristics, the amount of cholera toxin produced, their antibiograms, and the presence or absence of the pTLC plasmid sequence. DNA fingerprinting analysis by arbitrarily primed PCR, ribotyping, and a pulsed-field gel electrophoresis method classified the toxigenic strains into 3, 7, and 10 types, respectively. The relatedness of these toxigenic strains to clinical strains isolated in other countries and from international travelers was examined by using a dendrogram constructed from the pulsed-field gel electrophoresis profiles. The results of the examination of the antibiogram and the possession of the toxin-linked cryptic plasmid were consistent with the dendrogram-based relatedness: the O139 strains isolated from Malaysian seafoods could be separated into two groups that appear to have been introduced from the Bengal area independently. The rough strains of Malaysian seafood origin formed one group and belonged to a cluster unique to the Thailand-Malaysia-Laos region, and this group may have persisted in this area for a long period. The single O1 Ogawa strain detected in Malaysian seafood appears to have an origin and route of introduction different from those of the O139 and the rough strains.


2021 ◽  
Vol 14 (11) ◽  
Author(s):  
Farzad Mohamadi ◽  
Jalil Vand Yousefi ◽  
Naser Harzandi ◽  
Sobhan Ghafourian

Background: Due to the importance of identifying the source of infectious agents, different typing methods have been developed, among which the pulsed-field gel electrophoresis (PFGE) method is known as the gold standard for bacteria. Also, Enterococcus faecalis is classified as a nosocomial infection. Objectives: The current study aimed to identify the source of E. faecalis by the PFGE method. Methods: Bacteria were collected from all cases of urinary tract infections. Then, the identification process was performed. All isolates were evaluated for vancomycin resistance, and then PFGE was carried out. Results: The results of disk diffusion showed that 54% of the isolates showed resistance to vancomycin. Also, 4% of the isolates were intermediate, and 42% showed sensitivity to vancomycin. Afterwards, the PCR of the VanA gene was performed to confirm the results of disk diffusion. Thus, 48 out of 54 (88.8%) isolates had the VanA gene, and none of the four intermediate isolates had the VanA gene. Our results demonstrated that 54 isolates were vancomycin-resistant, and 50 different pulsotypes groups were identified. Conclusions: Our findings showed the isolates of E. faecalis were from different clonal lineages.


1999 ◽  
Vol 122 (3) ◽  
pp. 417-422 ◽  
Author(s):  
G. MARTINEZ ◽  
K. AHMED ◽  
C. H. ZHENG ◽  
K. WATANABE ◽  
K. OISHI ◽  
...  

Pulsed field gel electrophoresis (PFGE) of the genomic DNA of Moraxella catarrhalis was done in 172 strains isolated from sputum of patients with respiratory infections in Nagasaki (130 strains), Europe (14 strains), Thailand (6 strains), Uganda (3 strains), Bangladesh (5 strains) and Kuwait (14 strains). Restriction endonuclease with SmaI generated 4–16 DNA fragments ranging from 1000 kb to 24·25 kb and was classified into 31 major groups. It was found that there were wide variations of DNA restriction patterns of strains isolated from the same and different geographical areas. DNA restriction patterns of strains isolated in Nagasaki during the last 12 years showed dynamic changes of the predominant strains in each time period. We conclude from this study that PFGE is a suitable method to document interstrain variation in M. catarrhalis.


1999 ◽  
Vol 37 (2) ◽  
pp. 450-452 ◽  
Author(s):  
Hoang Vu-Thien ◽  
Carole Dulot ◽  
Didier Moissenet ◽  
Brigitte Fauroux ◽  
Antoine Garbarg-Chenon

Randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) for the analysis of 13 Moraxella catarrhalis isolates, 11 successive strains isolated from sputa of five children and 2 isolates obtained the same day from twins, were compared. RAPD and PFGE both yielded nine types from the 13 isolates, showing a chronic colonization with one strain in three patients and a successive colonization with different strains in two patients. The promising results obtained with RAPD should be confirmed with a larger number of strains, but RAPD seems as suitable as PFGE for the typing ofM. catarrhalis.


1995 ◽  
Vol 6 (3) ◽  
pp. 141-144 ◽  
Author(s):  
Susan M Davison ◽  
Donald E Low ◽  
Rene H Cruz ◽  
Danielle Beaulieu ◽  
Shelley R Scriver

Pulsed field gel electrophoresis (pfge) was used to compare 59 strains ofMoraxella catarrhalisto evaluatepfgefor the epidemiological typing of this organism.pfge-generated patterns were compared with those obtained by small fragment restriction enzyme analysis (rea) and species-specific probe hybridization. The strains used in the study were isolated from various geographic locations and included proven epidemiologically related strains.pfgeyielded more unique patterns thandna-dnahybridization – 30 versus 18, respectively – but fewer thanrea, which generated 45 unique patterns. Strains that demonstrated the samereapattern ordna-dnahybridization pattern did not always demonstrate the samepfgepattern. For example, in 23 epidemiologically unrelated strains that shared sixreapatterns,pfgedifferentiated the isolates into 12 patterns. Conversely, strains that demonstrated the samepfgepattern did not always demonstrate the samereapattern or hybridization pattern. For example, in 42 strains that shared 13pfgepatterns,readifferentiated the isolates into 31 patterns anddna-dnahybridization differentiated them into 16 patterns. However, compared withrea,pfgeyielded less complex patterns that were more easily comparable, and compared withdna-dnahybridization,pfgewas technically easier.


2000 ◽  
Vol 38 (2) ◽  
pp. 625-629 ◽  
Author(s):  
Hisakazu Yano ◽  
Mitsuko Suetake ◽  
Akio Kuga ◽  
Kazuhiko Irinoda ◽  
Ryoichi Okamoto ◽  
...  

To investigate how bacterial pathogens spread from child to child in a day care center, we monitored six children, two boys and four girls, born between August 1995 and November 1997, attending a day care center and analyzed nasopharyngeal samples from them using pulsed-field gel electrophoresis (PFGE). We obtained nasopharyngeal cultures from all of the affected children and almost all of the unaffected children between September 1998 and March 1999 after some children presented simultaneously with purulent rhinorrhea. Moreover, when a child was found to have acute otitis media, nasopharyngeal secretions from the child were independently cultured during treatment. During this period, 28 isolates of Moraxella catarrhalis, 13 ofStreptococcus pneumoniae, and 4 of Haemophilus influenzae were recovered. PFGE gave 8 patterns for M. catarrhalis, 10 for S. pneumoniae, and 1 for H. influenzae. PFGE patterns demonstrated spread of M. catarrhalis between children. However, each occurrence of clusters of infection with M. catarrhalis lasted 2 to 6 weeks, with a change in PFGE pattern between occurrences of clusters. The M. catarrhalis strain infecting each child also changed. Similarly, the S. pneumoniae strain in each child also changed. In contrast, infection with H. influenzaepersisted for about 3 months in an affected child.


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