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140 Subclonal variation of a cystic fibrosis (CF) epidemic strain of Pseudomonas aeruginosa
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Journal of Cystic Fibrosis 4 (2005) $34 $58
137 Preliminary results of a longitudinal study observing the impact of Staphylococcus aureus nasal carriage on colonization of the airways in young CF patients S. Ridder Schaphorn 1, C. Breit kopf 1, B. Ritzer reid 1, A. D/ibber s2, J. Hiiberle~, S. F a l k9, R K/isters, A. Schuster 4, E Ratjen ~, B. Sinha 1, G Peters 1, B.C. Kahl 1
~Med. Microbiology, 2Dep. Pediatrics, Uni~ Hosp. Muenster; SClemenshospitaI Muenster; 4Univ. Hosp. DuesseMorf and SEsser~ Germany A i m s : Nasal cmriage of Staphylococcus aureus represents a r i s k factor for subsequent infections in certain patient populations. It is not known, it" nasal S. aureus cmriage in CF patients represents a risk factor for subsequent colonisation and infection of the airways. Methods: Nasal (n 496) andtbroat swabs (n 578) or sputum (n 68) of 84 CF children younger than six years ( m e a n age 4.2 years) w e r e collected from 1999 to 2CO4. Two groups served as control (group 1, m e a n age 8.9 years; group 2, mean age 2.2 years). Clonal identity of S. aureus strains was determined by puls~l field gel electr ophoresis. Results: S. aureus was cultured in 54/84 children ( 6 1 % ) from the nares, in 68/84 (78 %) from the tbroat and in 9/13 (69 %) from the sputum. Nasal carriage of control group 1 and 2 w e r e 33 % (15/46) and 24 % (10/42), respectively, ha 10 patients, the nose was colonized first, w h i l e in 8 patients S. aureus was first cultured from the tbroat. Conclusions: The nasal colonization rate of CF patients is significantly higher than ha other children. T h e anterior nares seem to play an important role in the further colonization of the airways. Therefore, a n e w treatment strategy for the prevention or postponing of S. aureus airway infection in y o u n g CF patients should b e followed by implementing an eradication pr ceedure of the nasal S. aureus carriage w i t h topical mupir cein.
139 Prevalence of a cystic fibrosis (CF) epidemic sb'ain of Pseudomonas aeruginosa in Liverpool, U.K. S. Pmaagea 1, L. Humphrey s2, M.J. Walshaw ~, M.J. Ledson 3, C. Wins~maley1, C.A. Hart 1
iDepartrnent of Medical Microbiolog£ University of Liverpool, Liverpool, U.K, 2Microbiology Department, Royal arm Broadgreen University Hospitals NHS Trust, Liverpool, U.K, 3RegionaI Adult Cystic Fibrosis Unit, Carzliothoracic Centre, Liverpool, U.K.
This study was funded by the German Mukoviszidose e. k', project F04/03.
A n epidemic strain of /~ aeruginosa, the Liverpool epidemic strain (LES), is widespread among CF patients attending clinics ha Liverpool, United Kingdom. A i m s : Screening of all patients attending the Liverpool regional adult CF centre was undertaken w i t h regard to LES colonisation in order to introduce segregation of those w h o harbour LES. M e t h o d s : M u l t i p l e respiratory samples collected over a three year period from 173 patients w e r e tested for the presence o f / ~ aeruginosa and LES by at least one of the following methods: a) culture and PS21 PCR (specific for LES) o f / ~ aeruginosa strains, b) culture and R A P D typing o f / ~ aeruginosa strains, mad c) PS21 PCR directly on sputum. Results: One hundred mad forty n i n e patients (86%) w e r e colonised w i t h / ~ aeruginosa and of these 113 (76%) harboured LES. Twenty seven percent of the patients colonised w i t h LES harboured non epidemic/~ aeruginosa strains as well. T h e r e w e r e 13 cases of acquisition of LES during the study period. Ten of these patients became cbr onically infected with LES. Cases of superinfection w i t h LES and replacement of u n i q u e / ~ aeruginosa strains by LES w e r e detected. C o n d u s i o n s : This study confirms a very h i g h rate of colonisation w i t h LES in the Liverpool adult CF centre mad shows evidence of continuous spread of this strain w i t h i n the unit. R e g u l a r microbiological surveillance w i l l be required to show the effects of segregation of LES colonised from non~colonised patients, w h i c h was introduced during the second halt" of this study.
138 Increasing prevalence of stable Staphylococcus aureus (SA) small colony variant (SCV) phenotypes in CF pafients
140 Subclonal variation of a cystic fibrosis (CF) epidemic strain of Pseudomonas aeruginosa
G. Marglo 1, A. D'Alessandri 9, M. M e n t asti 1, E. Poggi 9, E. D i Marco 1, G. CangeIni 1, R M o r e l l i 1, C. B e m a r d i 1, R. Casciar d
S. Pmaagea l, J.E. Corkill2, C. Winst maley l, C.A. Hart l
iClinicaI Pathology Lab; 2CF Unit; Gaslini Hospital Genoa-Italy
iDepartrnent of Medical Microbiolog~ University of Liverpool, Liverpool, U.K, 2Microbiology Department, Royal arul Broadgreen University Hospitals NHS Trust, Liverpool, U.K.
B a e k g m t m d SASCVs are intr acellular, anxotr ophes, slow g r o w i n g subpopulations w i t h reduced antibiotic susceptibility, w h i c h can cause persisting and relapsing infections in CF. A i m s Determine prevalence and antibiotic susceptibility of SASCVs in CF patients (pts) attending the Genoa Centre in 2 0 0 4 and the possible effect of antibiotic treatments in selection of these strains. M e t h o d s SA mad S A S C V w e r e recovered from sputa and tbroat swabs in Mamfitol Salt Agar after 4 8 h at 35°C and identified by latex agglutination, coagulase tube, API Staph system and specific SA nuc gene. Auxotrophism assessed by thymidine (T), h e m i n mad menadione disks. M e t i c i l l i n msceptibility confirmed by mecA gene. Antibiotic treatment courses before S A S C V recovering, FEV1 and B M I w e r e assessed. Results 25/199 pts had stable SASCV. SCV+normal SA w e r e recovered from 14 pts, 11 pts harboured only SCV. 21pts w i t h S A S C V w e r e colonised also by t~aeruginosa. SCVs w e r e T d e p e n d e n t (92%), h e m i n dependent (4%) and meticillin resistant (36%); all T d e p e n d e n t SCVs w e r e cotrimoxazole (SXT) resistant. Before first S A S C V recovering, pts h a d therapy courses w i t h : 11/25 nebulised aminogligosides+various, 8/25 n e b u l i s e d aminoglicosides+SXT and 4/25 SXT+various. Pts receiving TOBI w e r e 60%. N o significant differences in FEV1 and B M I before and after SASCV colonization w e r e found. Conclusions Our data show an increasing occturence of SASCV ha 2 ~ ) 4 (12.5%) compared w i t h previous years (7% in 2CO)); this finding could be related to the antibiotic pressure and extensive use of T O B I and SXT. S A S C V could spread a m o n g pts, so accurate microbiological protocols for detection and antibiotics susceptibility testing are critical.
A highly transmissible, epidemic strain of/~ aeruginosa is widespread among CF patients attending clinics in Liverpool, U.K. A i m s : W e studied the emergence of clonal variants of this Liverpool epidemic strain (LES) over t i m e in chronically infected patients. M e t h o d s : Forty LES isolates from seven patients w e r e typed by macrorestriction analysis of SpeI and XbaI digested DNA by PFGE. A total of 4 to 8 isolates from each patient w e r e studied, collected at three different times during their colonisation (1995 2003). Macrorestriction fragment patterns w e r e compared w i t h the first LES in our collection isolated in 1988. A l l isolates w e r e tested positive w i t h a PCR assay specific for LES. Results: P F G E analysis of SpeI and XbaI digests revealed 9 and 7 clonal variants of LES respectively. T h e r e was overall good correlation between SpeI and XbaI fragment patterns, however digestion w i t h SpeI was more discriminatory in this study. A l l patients had clonal variants of LES (range 2 4). The variant w i t h identical fragment pattern to the 1988 isolate was the most common and was isolated from all but one patient at least once. Emergence of clonal variants in individual patients appeared to occur at random and did not persist tbroughout the study period in most of them. No asscciation between clonal variants and phenotype or antibiotic sensitivity pattem of the isolates was observed. Conclusions: Clonal variants of LES are common and w e r e detected ha all the patients. This study confirms that genomic diversity and evolution of a clonal lineage as indicated by subtle band shifts of the macr orestriction fragment pattern is common in chronically infected CF patients.
Journal of Cystic Fibrosis 4 (2005) $34 $58
137 Preliminary results of a longitudinal study observing the impact of Staphylococcus aureus nasal carriage on colonization of the airways in young CF patients S. Ridder Schaphorn 1, C. Breit kopf 1, B. Ritzer reid 1, A. D/ibber s2, J. Hiiberle~, S. F a l k9, R K/isters, A. Schuster 4, E Ratjen ~, B. Sinha 1, G Peters 1, B.C. Kahl 1
~Med. Microbiology, 2Dep. Pediatrics, Uni~ Hosp. Muenster; SClemenshospitaI Muenster; 4Univ. Hosp. DuesseMorf and SEsser~ Germany A i m s : Nasal cmriage of Staphylococcus aureus represents a r i s k factor for subsequent infections in certain patient populations. It is not known, it" nasal S. aureus cmriage in CF patients represents a risk factor for subsequent colonisation and infection of the airways. Methods: Nasal (n 496) andtbroat swabs (n 578) or sputum (n 68) of 84 CF children younger than six years ( m e a n age 4.2 years) w e r e collected from 1999 to 2CO4. Two groups served as control (group 1, m e a n age 8.9 years; group 2, mean age 2.2 years). Clonal identity of S. aureus strains was determined by puls~l field gel electr ophoresis. Results: S. aureus was cultured in 54/84 children ( 6 1 % ) from the nares, in 68/84 (78 %) from the tbroat and in 9/13 (69 %) from the sputum. Nasal carriage of control group 1 and 2 w e r e 33 % (15/46) and 24 % (10/42), respectively, ha 10 patients, the nose was colonized first, w h i l e in 8 patients S. aureus was first cultured from the tbroat. Conclusions: The nasal colonization rate of CF patients is significantly higher than ha other children. T h e anterior nares seem to play an important role in the further colonization of the airways. Therefore, a n e w treatment strategy for the prevention or postponing of S. aureus airway infection in y o u n g CF patients should b e followed by implementing an eradication pr ceedure of the nasal S. aureus carriage w i t h topical mupir cein.
139 Prevalence of a cystic fibrosis (CF) epidemic sb'ain of Pseudomonas aeruginosa in Liverpool, U.K. S. Pmaagea 1, L. Humphrey s2, M.J. Walshaw ~, M.J. Ledson 3, C. Wins~maley1, C.A. Hart 1
iDepartrnent of Medical Microbiolog£ University of Liverpool, Liverpool, U.K, 2Microbiology Department, Royal arm Broadgreen University Hospitals NHS Trust, Liverpool, U.K, 3RegionaI Adult Cystic Fibrosis Unit, Carzliothoracic Centre, Liverpool, U.K.
This study was funded by the German Mukoviszidose e. k', project F04/03.
A n epidemic strain of /~ aeruginosa, the Liverpool epidemic strain (LES), is widespread among CF patients attending clinics ha Liverpool, United Kingdom. A i m s : Screening of all patients attending the Liverpool regional adult CF centre was undertaken w i t h regard to LES colonisation in order to introduce segregation of those w h o harbour LES. M e t h o d s : M u l t i p l e respiratory samples collected over a three year period from 173 patients w e r e tested for the presence o f / ~ aeruginosa and LES by at least one of the following methods: a) culture and PS21 PCR (specific for LES) o f / ~ aeruginosa strains, b) culture and R A P D typing o f / ~ aeruginosa strains, mad c) PS21 PCR directly on sputum. Results: One hundred mad forty n i n e patients (86%) w e r e colonised w i t h / ~ aeruginosa and of these 113 (76%) harboured LES. Twenty seven percent of the patients colonised w i t h LES harboured non epidemic/~ aeruginosa strains as well. T h e r e w e r e 13 cases of acquisition of LES during the study period. Ten of these patients became cbr onically infected with LES. Cases of superinfection w i t h LES and replacement of u n i q u e / ~ aeruginosa strains by LES w e r e detected. C o n d u s i o n s : This study confirms a very h i g h rate of colonisation w i t h LES in the Liverpool adult CF centre mad shows evidence of continuous spread of this strain w i t h i n the unit. R e g u l a r microbiological surveillance w i l l be required to show the effects of segregation of LES colonised from non~colonised patients, w h i c h was introduced during the second halt" of this study.
138 Increasing prevalence of stable Staphylococcus aureus (SA) small colony variant (SCV) phenotypes in CF pafients
140 Subclonal variation of a cystic fibrosis (CF) epidemic strain of Pseudomonas aeruginosa
G. Marglo 1, A. D'Alessandri 9, M. M e n t asti 1, E. Poggi 9, E. D i Marco 1, G. CangeIni 1, R M o r e l l i 1, C. B e m a r d i 1, R. Casciar d
S. Pmaagea l, J.E. Corkill2, C. Winst maley l, C.A. Hart l
iClinicaI Pathology Lab; 2CF Unit; Gaslini Hospital Genoa-Italy
iDepartrnent of Medical Microbiolog~ University of Liverpool, Liverpool, U.K, 2Microbiology Department, Royal arul Broadgreen University Hospitals NHS Trust, Liverpool, U.K.
B a e k g m t m d SASCVs are intr acellular, anxotr ophes, slow g r o w i n g subpopulations w i t h reduced antibiotic susceptibility, w h i c h can cause persisting and relapsing infections in CF. A i m s Determine prevalence and antibiotic susceptibility of SASCVs in CF patients (pts) attending the Genoa Centre in 2 0 0 4 and the possible effect of antibiotic treatments in selection of these strains. M e t h o d s SA mad S A S C V w e r e recovered from sputa and tbroat swabs in Mamfitol Salt Agar after 4 8 h at 35°C and identified by latex agglutination, coagulase tube, API Staph system and specific SA nuc gene. Auxotrophism assessed by thymidine (T), h e m i n mad menadione disks. M e t i c i l l i n msceptibility confirmed by mecA gene. Antibiotic treatment courses before S A S C V recovering, FEV1 and B M I w e r e assessed. Results 25/199 pts had stable SASCV. SCV+normal SA w e r e recovered from 14 pts, 11 pts harboured only SCV. 21pts w i t h S A S C V w e r e colonised also by t~aeruginosa. SCVs w e r e T d e p e n d e n t (92%), h e m i n dependent (4%) and meticillin resistant (36%); all T d e p e n d e n t SCVs w e r e cotrimoxazole (SXT) resistant. Before first S A S C V recovering, pts h a d therapy courses w i t h : 11/25 nebulised aminogligosides+various, 8/25 n e b u l i s e d aminoglicosides+SXT and 4/25 SXT+various. Pts receiving TOBI w e r e 60%. N o significant differences in FEV1 and B M I before and after SASCV colonization w e r e found. Conclusions Our data show an increasing occturence of SASCV ha 2 ~ ) 4 (12.5%) compared w i t h previous years (7% in 2CO)); this finding could be related to the antibiotic pressure and extensive use of T O B I and SXT. S A S C V could spread a m o n g pts, so accurate microbiological protocols for detection and antibiotics susceptibility testing are critical.
A highly transmissible, epidemic strain of/~ aeruginosa is widespread among CF patients attending clinics in Liverpool, U.K. A i m s : W e studied the emergence of clonal variants of this Liverpool epidemic strain (LES) over t i m e in chronically infected patients. M e t h o d s : Forty LES isolates from seven patients w e r e typed by macrorestriction analysis of SpeI and XbaI digested DNA by PFGE. A total of 4 to 8 isolates from each patient w e r e studied, collected at three different times during their colonisation (1995 2003). Macrorestriction fragment patterns w e r e compared w i t h the first LES in our collection isolated in 1988. A l l isolates w e r e tested positive w i t h a PCR assay specific for LES. Results: P F G E analysis of SpeI and XbaI digests revealed 9 and 7 clonal variants of LES respectively. T h e r e was overall good correlation between SpeI and XbaI fragment patterns, however digestion w i t h SpeI was more discriminatory in this study. A l l patients had clonal variants of LES (range 2 4). The variant w i t h identical fragment pattern to the 1988 isolate was the most common and was isolated from all but one patient at least once. Emergence of clonal variants in individual patients appeared to occur at random and did not persist tbroughout the study period in most of them. No asscciation between clonal variants and phenotype or antibiotic sensitivity pattem of the isolates was observed. Conclusions: Clonal variants of LES are common and w e r e detected ha all the patients. This study confirms that genomic diversity and evolution of a clonal lineage as indicated by subtle band shifts of the macr orestriction fragment pattern is common in chronically infected CF patients.