Development of a DNA microarray to identify the Streptococcus pneumoniae serotypes contained in the 23-valent pneumococcal polysaccharide vaccine and closely related serotypes.
Streptococcus pneumoniae is a major worldwide human pathogen. This investigation has developed a reliable and accurate DNA microarray method for inter-species differentiation of S. pneumoniae and intra-species differentiation of the 23 groups of S. pneumoniae including serotypes represented in the 23-valent pneumococcal vaccine and the other 20 closely related serotypes. In addition to 16S rDNA probes, serotype- or serogroup-specific probes targeting the capsular polysaccharide synthesis (cps) genes, wzy or capA were generated. We adopted a two-step multiplex PCR to improve the sensitivity of detection to a level of 10(5) cfu/ml in pure culture or 50 ng DNA. A total of 169 isolates (from China, Australia, Canada and New Zealand) including 147 belonging to 23-valent vaccine and closely related serotypes of S. pneumoniae, 11 belonging to other serotypes and 11 of different species commonly isolated from respiratory tract were tested to verify the method. The DNA microarray method developed provides a sensitive means to rapidly identify the members of the most common S. pneumoniae serotypes in patients and to monitor their distribution in different patient groups and geographic locations. Such information is needed for disease surveillance and to monitor vaccine efficacy.
Wang
Quan
Q
TEDA School of Biological Sciences and Biotechnology, Nankai University, 23# HongDa Street, TEDA, Tianjin 300457, China.
Wang
Min
M
Kong
Fanrong
F
Gilbert
Gwendolyn L
GL
Cao
Boyang
B
Wang
Lei
L
Feng
Lu
L
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008856:Microscopy, Fluorescence; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011008:Pneumococcal Infections / Q000175:diagnosis*; D022242:Pneumococcal Vaccines / Q000145:classification* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000379:methods; D013296:Streptococcus pneumoniae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
C414006:23-valent pneumococcal capsular polysaccharide vaccine; D001426:Bacterial Proteins; C500439:CpsA protein, Streptococcus pneumoniae; D004269:DNA, Bacterial; D022242:Pneumococcal Vaccines; C474640:CpsD protein, Streptococcus pneumoniae; D011505:Protein-Tyrosine Kinases; C500440:CpsB protein, Streptococcus pneumoniae; D017027:Protein Tyrosine Phosphatases