Worldwide Lineages of Clinical Pneumococci in a Japanese Teaching Hospital Identified by DiversiLab System.
Pneumococcal Molecular Epidemiology Network (PMEN) clones are representatives of worldwide-spreading pathogens. DiversiLab system, a repetitive PCR system, has been proposed as a less labor-and time-intensive genotyping platform alternative to conventional methods. However, the utility and analysis parameters of DiversiLab for identifying worldwide lineages was not established. To evaluate and optimize the performance of DiversiLab for identifying worldwide pneumococcal lineages, we examined 245 consecutive isolates of clinical Streptococcus pneumoniae from all age-group patients at a teaching hospital in Japan. The capsular swelling reaction of all isolates yielded 24 different serotypes. Intensive visual observation (VO) of DiversiLab band pattern difference divided all isolates into 73 clusters. Multilocus sequence typing (MLST) of representative 73 isolates from each VO cluster yielded 51 different STs. Among them, PMEN-related lineages accounted for 63% (46/73). Although the serotype of PMEN-related isolates was identical to that of the original PMEN clone in 70% (32/46), CC156-related PMEN lineages, namely Greece(6B)-22 and Colombia(23F)-26, harbored various capsular types discordant to the original PMEN clones. Regarding automated analysis, genotyping by extended Jaccard (XJ) with a 75% similarity index cutoff (SIC) showed the highest correlation with serotyping (adjusted Rand's coefficient, 0.528). Elevating the SIC for XJ to 85% increased the discriminatory power sufficient for distinguishing two major PMEN-related isolates of Taiwan(19F)-14 and Netherlands(3)-31. These results demonstrated a potential utility of DiversiLab for identifying worldwide lineage of pneumococcus. An optimized parameters of automated analysis should be useful especially for comparison for reference strains by "identification" function of DiversiLab.
Department of Microbiology and Infectious Diseases, Toho University School of Medicine, Tokyo, Japan; Department of Clinical Laboratory, Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital, Tokyo, Japan.
Kiyoshi
K
Kashiwaya
Department of Microbiology and Infectious Diseases, Toho University School of Medicine, Tokyo, Japan; Central Laboratory Division, Akita University Hospital, Akita, Japan. Electronic address: sagatomoo@gmail.com.
Tomoo
T
Saga
Department of Microbiology and Infectious Diseases, Toho University School of Medicine, Tokyo, Japan.
Yoshikazu
Y
Ishii
Department of Microbiology and Infectious Diseases, Toho University School of Medicine, Tokyo, Japan.
Ryuji
R
Sakata
Clinical Laboratory Department, Toho University Omori Medical Center, Tokyo, Japan.
Morihiro
M
Iwata
Division of Infection Control, Toho University Omori Medical Center, Tokyo, Japan.
Sadako
S
Yoshizawa
Department of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan.
Bin
B
Chang
Department of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan.
Makoto
M
Ohnishi
Department of Microbiology and Infectious Diseases, Toho University School of Medicine, Tokyo, Japan; Clinical Laboratory Department, Toho University Omori Medical Center, Tokyo, Japan; Division of Infection Control, Toho University Omori Medical Center, Tokyo, Japan.
Kazuhiro
K
Tateda
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D005838:Genotype; D006784:Hospitals, Teaching; D006801:Humans; D007223:Infant; D007564:Japan; D008875:Middle Aged; D058885:Multilocus Sequence Typing / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D012703:Serotyping; D013296:Streptococcus pneumoniae / Q000145:classification* / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D055815:Young Adult
Automated analysis; DiversiLab; Multilocus sequence typing (MLST); Repetitive PCR (rep-PCR); Serotypes; Streptococcus pneumoniae