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Persistent Bordetella petrii infection related to bone fractures.

Kwon SS, Kim JO, Kim KH, Jeong SH, Lee K - Ann Lab Med (2016)

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, Korea.

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Dear Editor, Bordetella, of the family Alcaligenaceae, is a genus containing gram-negative coccoid rod species... To date, nine species have been assigned to the genus Bordetella... The wound culture revealed a gram-negative rod identified as Achromobacter denitrificans by using a VITEK 2 GNI card (bioMérieux, Marcy l'Etoile, France)... Smear preparation of these colonies showed short gram-negative rods... Subculture on triple-sugar iron slant agar showed that the bacteria did not ferment glucose or produce H2S... However, all three clinical isolates were resistant to cefepime and ciprofloxacin, while the type strain was susceptible to both drugs (Table 1)... Cephalexin was used as empirical therapy and changed to levofloxacin following AST results... Former clinical isolates were identified as A. denitrificans, B. bronchiseptica, and Alcaligenes species in hospitals A and B... However, all these isolates are speculated to be B. petrii because commercially available cards misidentified our three clinical isolates and type strain DSM 12804 as A. denitrificans and Burkholderia species... However, MALDI-TOF MS identified the isolates correctly with a high score... When bacterial culture results show discrepancies among different bacterial identification methods and the results include members of Alcaligenaceae, B. petrii should be suspected and confirmed by using MALDI-TOF MS or 16S rRNA sequencing... Long-lasting B. petrii infections have been reported previously... Chronic infections, despite continuous use of antibiotics, suggest the clinical difficulty of treating B. petrii... Bone-related infections are associated with reduced effects of antimicrobial therapy, which possibly contribute to persistent B. petrii infection.

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Related in: MedlinePlus

Pulsed field gel electrophoresis (PFGE) was performed by using the CHEF-DRII System (Bio-Rad, Hercules, CA, USA) at 6 V/cm for 20 hr at 10℃, with initial and final pulse times of 0.5 sec and 30 sec, respectively. XbaI was the restriction enzyme used. All three clinical isolates from our hospital (hospital C) exhibited 100% identical PFGE banding patterns. This finding suggests that sequentially identified B. petrii were the same strains. Lane C1-3, Bordetella petrii isolated from patient's wound; ATCC, ATCC DSM 12804.
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Figure 1: Pulsed field gel electrophoresis (PFGE) was performed by using the CHEF-DRII System (Bio-Rad, Hercules, CA, USA) at 6 V/cm for 20 hr at 10℃, with initial and final pulse times of 0.5 sec and 30 sec, respectively. XbaI was the restriction enzyme used. All three clinical isolates from our hospital (hospital C) exhibited 100% identical PFGE banding patterns. This finding suggests that sequentially identified B. petrii were the same strains. Lane C1-3, Bordetella petrii isolated from patient's wound; ATCC, ATCC DSM 12804.

Mentions: Deep wound culture yielded medium to large grayish and convex colonies that were non-hemolytic on sheep blood agar plate (BAP), and colorless, opaque pinpoint-sized colonies on MacConkey agar plate (isolate C1). Two serial follow-up cultures revealed the same species (isolates C2 and C3). Smear preparation of these colonies showed short gram-negative rods. Subculture on triple-sugar iron slant agar showed that the bacteria did not ferment glucose or produce H2S. Isolates exhibited an umbrella-shaped growth pattern when cultured in motility-indole-ornithine semisolid agar, suggesting that the microorganism is strictly aerophilic and motile. API 20NE (bioMérieux) and VITEK 2 GNI cards identified the isolates as A. denitrificans and Burkholderia species, respectively. However, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS; Bruker Daltonics, Billerica, MA, USA) identified them as B. petrii with a 2.089 score. The isolates were finally confirmed as B. petrii on the basis of partial sequences of the 16S rRNA gene, which showed 100% identity to those of DSM 12804, the type strain of B. petrii, when analyzed by BLASTn (http://www.ncbi.nlm.nih.gov/BLAST). XbaI-macrorestriction analysis was performed by using pulse field gel electrophoresis (PFGE) (CHEF-DRII System, Bio-Rad, Hercules, CA, USA), as described previously [9]. All three clinical isolates exhibited identical PFGE banding patterns (Fig. 1).


Persistent Bordetella petrii infection related to bone fractures.

Kwon SS, Kim JO, Kim KH, Jeong SH, Lee K - Ann Lab Med (2016)

Pulsed field gel electrophoresis (PFGE) was performed by using the CHEF-DRII System (Bio-Rad, Hercules, CA, USA) at 6 V/cm for 20 hr at 10℃, with initial and final pulse times of 0.5 sec and 30 sec, respectively. XbaI was the restriction enzyme used. All three clinical isolates from our hospital (hospital C) exhibited 100% identical PFGE banding patterns. This finding suggests that sequentially identified B. petrii were the same strains. Lane C1-3, Bordetella petrii isolated from patient's wound; ATCC, ATCC DSM 12804.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4697348&req=5

Figure 1: Pulsed field gel electrophoresis (PFGE) was performed by using the CHEF-DRII System (Bio-Rad, Hercules, CA, USA) at 6 V/cm for 20 hr at 10℃, with initial and final pulse times of 0.5 sec and 30 sec, respectively. XbaI was the restriction enzyme used. All three clinical isolates from our hospital (hospital C) exhibited 100% identical PFGE banding patterns. This finding suggests that sequentially identified B. petrii were the same strains. Lane C1-3, Bordetella petrii isolated from patient's wound; ATCC, ATCC DSM 12804.
Mentions: Deep wound culture yielded medium to large grayish and convex colonies that were non-hemolytic on sheep blood agar plate (BAP), and colorless, opaque pinpoint-sized colonies on MacConkey agar plate (isolate C1). Two serial follow-up cultures revealed the same species (isolates C2 and C3). Smear preparation of these colonies showed short gram-negative rods. Subculture on triple-sugar iron slant agar showed that the bacteria did not ferment glucose or produce H2S. Isolates exhibited an umbrella-shaped growth pattern when cultured in motility-indole-ornithine semisolid agar, suggesting that the microorganism is strictly aerophilic and motile. API 20NE (bioMérieux) and VITEK 2 GNI cards identified the isolates as A. denitrificans and Burkholderia species, respectively. However, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS; Bruker Daltonics, Billerica, MA, USA) identified them as B. petrii with a 2.089 score. The isolates were finally confirmed as B. petrii on the basis of partial sequences of the 16S rRNA gene, which showed 100% identity to those of DSM 12804, the type strain of B. petrii, when analyzed by BLASTn (http://www.ncbi.nlm.nih.gov/BLAST). XbaI-macrorestriction analysis was performed by using pulse field gel electrophoresis (PFGE) (CHEF-DRII System, Bio-Rad, Hercules, CA, USA), as described previously [9]. All three clinical isolates exhibited identical PFGE banding patterns (Fig. 1).

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, Korea.

AUTOMATICALLY GENERATED EXCERPT
Please rate it.

Dear Editor, Bordetella, of the family Alcaligenaceae, is a genus containing gram-negative coccoid rod species... To date, nine species have been assigned to the genus Bordetella... The wound culture revealed a gram-negative rod identified as Achromobacter denitrificans by using a VITEK 2 GNI card (bioMérieux, Marcy l'Etoile, France)... Smear preparation of these colonies showed short gram-negative rods... Subculture on triple-sugar iron slant agar showed that the bacteria did not ferment glucose or produce H2S... However, all three clinical isolates were resistant to cefepime and ciprofloxacin, while the type strain was susceptible to both drugs (Table 1)... Cephalexin was used as empirical therapy and changed to levofloxacin following AST results... Former clinical isolates were identified as A. denitrificans, B. bronchiseptica, and Alcaligenes species in hospitals A and B... However, all these isolates are speculated to be B. petrii because commercially available cards misidentified our three clinical isolates and type strain DSM 12804 as A. denitrificans and Burkholderia species... However, MALDI-TOF MS identified the isolates correctly with a high score... When bacterial culture results show discrepancies among different bacterial identification methods and the results include members of Alcaligenaceae, B. petrii should be suspected and confirmed by using MALDI-TOF MS or 16S rRNA sequencing... Long-lasting B. petrii infections have been reported previously... Chronic infections, despite continuous use of antibiotics, suggest the clinical difficulty of treating B. petrii... Bone-related infections are associated with reduced effects of antimicrobial therapy, which possibly contribute to persistent B. petrii infection.

No MeSH data available.


Related in: MedlinePlus