Borchardt Stephanie M, Foxman Betsy, Chaffin Donald O, Rubens Craig E, Tallman Patricia A, Manning Shannon D, Baker Carol J, Marrs Carl F
Department of Epidemiology, School of Public Health, University of Michigan, Ann Arbor, Michigan 48109, USA.
J Clin Microbiol. 2004 Jan;42(1):146-50. doi: 10.1128/JCM.42.1.146-150.2004.
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.
B族链球菌(GBS)(无乳链球菌)是新生儿和婴儿败血症及脑膜炎以及孕妇、非孕妇、可能免疫功能低下的成年人和老年人侵袭性疾病的主要病因。基于荚膜多糖抗原已描述了9种GBS血清型。侵袭性和定植性分离株的血清型分布在儿童和成人人群中有所不同,并且随时间发生了变化。因此,定期监测GBS血清型分布对于确保正确制定和应用适合人类使用的GBS疫苗以及检测新血清型的出现是必要的。自20世纪90年代中期以来,无法通过标准血清学方法分型的GBS分离株比例有所增加,这就需要更灵敏的分型方法。我们描述了一种分型方法,该方法使用DNA斑点印迹杂交,其探针由PCR从GBS血清型Ia、Ib和II至VIII的荚膜基因产生。设计PCR引物以扩增GBS荚膜基因的型特异性序列。从PCR产物构建基因探针,并根据杂交图谱对分离株进行分类。总共306株先前已血清分型的侵袭性和定植性分离株用于比较我们的斑点印迹荚膜分型(DBCT)鉴定方法与兰斯菲尔德血清分型(LS)。99%(306株中的303株)的分离株被指定了斑点印迹荚膜型,而306株分离株中的273株(89%)被指定了兰斯菲尔德血清型。两种方法之间的总体一致性为95%(两种方法均可分型的270株分离株中的256株)。我们得出结论,DBCT方法是常用LS方法的一种特异性且有用的替代方法。