title: Could the DiversiLab® semi-automated repetitive-sequence-based PCR be an acceptable technique for typing isolates of Pseudomonas aeruginosa? An answer from our experience and a review of the literature.
abstract: Recently the DiversiLab® (DL) system (bioMérieux) was developed as an automated platform that uses repetitive element polymerase chain reaction (rep-PCR) technology for standardized, reproducible DNA fingerprinting of bacteria. The purpose of this study was to evaluate the usefulness of DL rep-PCR for typing Pseudomonas aeruginosa isolates. The performance of DL rep-PCR was compared with that of pulsed-field gel electrophoresis (PFGE) in a prospective multicenter study of patients with ventilator-associated pneumonia due to P. aeruginosa, conducted in 3 intensive care units over a 31-month period. In total, 203 P. aeruginosa isolates from 66 patients, from whom at least 2 consecutive respiratory samples each were collected more than 48 h apart, were typed using DL rep-PCR. Forty isolates (corresponding to 20 patients) were also typed using PFGE of SpeI-digested DNA. The typeability was 100% with DL rep-PCR and 95% with PFGE. The discriminatory power was close for DL rep-PCR and for PFGE (Simpson's diversity indices of 0.901 and 0.947, respectively). Insufficient agreement between DL rep-PCR and PFGE typing results was observed for the 40 selected isolates (adjusted Rand coefficient of 0.419), mostly due to isolates of the same DL rep-PCR type but of different PFGE types (adjusted Wallace coefficients of 0.306 for DL rep-PCR with PFGE, and of 0.667 for PFGE with DL rep-PCR). Considered together with published data, DL rep-PCR results should be interpreted with caution for the investigation of outbreaks caused by P. aeruginosa and evaluated in conjunction with epidemiological data.
authors.affiliation: a PRES Sorbonne Universités, UPMC Univ Paris 06, INSERM, U1135, Centre for Immunology and Microbial Infections, team 13, F-75013 Paris, France.|b AP-HP, Hôpital Pitié-Salpêtrière, Bactériologie-Hygiène, F-75013 Paris, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Brossier
authors.affiliation: b AP-HP, Hôpital Pitié-Salpêtrière, Bactériologie-Hygiène, F-75013 Paris, France.
authors.forename: Maïté
authors.initials: M
authors.lastname: Micaelo
authors.affiliation: c AP-HP, Hôpital Pitié-Salpêtrière, Réanimation Médicale, F-75013 Paris, France.
authors.forename: Charles-Edouard
authors.initials: CE
authors.lastname: Luyt
authors.affiliation: d AP-HP, Hôpital Pitié-Salpêtrière, Réanimation Polyvalente, Département d'Anesthésie-Réanimation, F-75013 Paris, France.
authors.forename: Qin
authors.initials: Q
authors.lastname: Lu
authors.affiliation: c AP-HP, Hôpital Pitié-Salpêtrière, Réanimation Médicale, F-75013 Paris, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Chastre
authors.affiliation: d AP-HP, Hôpital Pitié-Salpêtrière, Réanimation Polyvalente, Département d'Anesthésie-Réanimation, F-75013 Paris, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Arbelot
authors.affiliation: c AP-HP, Hôpital Pitié-Salpêtrière, Réanimation Médicale, F-75013 Paris, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Trouillet
authors.affiliation: c AP-HP, Hôpital Pitié-Salpêtrière, Réanimation Médicale, F-75013 Paris, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Combes
authors.affiliation: d AP-HP, Hôpital Pitié-Salpêtrière, Réanimation Polyvalente, Département d'Anesthésie-Réanimation, F-75013 Paris, France.
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Rouby
authors.affiliation: a PRES Sorbonne Universités, UPMC Univ Paris 06, INSERM, U1135, Centre for Immunology and Microbial Infections, team 13, F-75013 Paris, France.|b AP-HP, Hôpital Pitié-Salpêtrière, Bactériologie-Hygiène, F-75013 Paris, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Jarlier
authors.affiliation: a PRES Sorbonne Universités, UPMC Univ Paris 06, INSERM, U1135, Centre for Immunology and Microbial Infections, team 13, F-75013 Paris, France.|b AP-HP, Hôpital Pitié-Salpêtrière, Bactériologie-Hygiène, F-75013 Paris, France.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Aubry
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D015373:Bacterial Typing Techniques / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D011014:Pneumonia / Q000175:diagnosis / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D011552:Pseudomonas Infections / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D012091:Repetitive Sequences, Nucleic Acid
keywords: Pseudomonas aeruginosa; bacterial typing; pulsed-field gel electrophoresis; rep-PCR; typage bactérien; électrophorèse sur gel en champ pulsé