title: Comparison of DNA dot blot hybridization and lancefield capillary precipitin methods for group B streptococcal capsular typing.
abstract: Group B streptococci (GBS) (Streptococcus agalactiae) are a major cause of sepsis and meningitis in neonates and infants and of invasive disease in pregnant women, nonpregnant, presumably immunocompromised adults, and the elderly. Nine GBS serotypes based on capsular polysaccharide antigens have been described. The serotype distributions among invasive and colonizing isolates differ between pediatric and adult populations and have changed over time. Thus, periodic monitoring of GBS serotype distributions is necessary to ensure the proper formulation and application of an appropriate GBS vaccine for human use and to detect the emergence of novel serotypes. Since the mid-1990s, the proportion of GBS isolates that are nontypeable by standard serologic methods has increased, creating a need for more sensitive typing methods. We describe a typing method that uses DNA dot blot hybridization with probes generated by PCR from the GBS capsular genes for serotypes Ia, Ib, and II to VIII. PCR primers were designed to amplify type-specific GBS capsular gene sequences. Gene probes were constructed from the PCR products and used to classify isolates based on hybridization profiles. A total of 306 previously serotyped invasive and colonizing isolates were used to compare our dot blot capsular typing (DBCT) identification method with Lancefield serotyping (LS). A dot blot capsular type was assigned to 99% (303 of 306) of the isolates, whereas 273 of 306 isolates (89%) were assigned a Lancefield serotype. The overall agreement between the methods was 95% (256 of 270 isolates typeable by both methods). We conclude that the DBCT method is a specific and useful alternative to the commonly used LS method.
authors.affiliation: Department of Epidemiology, School of Public Health, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Borchardt
authors.forename: Betsy
authors.initials: B
authors.lastname: Foxman
authors.forename: Donald O
authors.initials: DO
authors.lastname: Chaffin
authors.forename: Craig E
authors.initials: CE
authors.lastname: Rubens
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Tallman
authors.forename: Shannon D
authors.initials: SD
authors.lastname: Manning
authors.forename: Carol J
authors.initials: CJ
authors.lastname: Baker
authors.forename: Carl F
authors.initials: CF
authors.lastname: Marrs
mesh_terms: D016667:Bacterial Capsules / Q000145:classification*; D015373:Bacterial Typing Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012680:Sensitivity and Specificity; D012703:Serotyping; D013292:Streptococcus agalactiae / Q000145:classification* / Q000235:genetics
chemical_list: D004269:DNA, Bacterial