Beall B, Gherardi G, Facklam R R, Hollingshead S K
Respiratory Diseases Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Clin Microbiol. 2000 Oct;38(10):3663-9. doi: 10.1128/JCM.38.10.3663-3669.2000.
In a recent genotypic survey of beta-lactam-resistant pneumococci recovered in different areas of United States during 1997, eight clonal types that each represented 3 to 40 isolates accounted for 134 of 144 isolates (G. Gherardi, C. Whitney, R. Facklam, and B. Beall, J. Infect. Dis. 181:216-229, 2000). We determined the degree of pspA gene diversity among these 134 isolates and for 11 previously characterized internationally disseminated multiresistant strains. Thirty-four different pspA restriction profiles were determined for an amplicon encompassing the variable portion of the structural gene that encodes the surface-exposed domain of PspA and a variable-length proline-rich putative cell wall-associated domain. These restriction profiles closely correlated with those of 33 different pspA sequence types of an approximately 230-residue region corresponding to residues 182 to 410 of the strain Rx1 PspA. These residues encompass a 100-residue clade-defining region known to contain cross-protective epitopes for which 17 sequence types were found. Distinct, conserved pspA sequence types were found for the majority of strains within seven of the eight U.S. clonal types assessed, while one pulsed-field gel electrophoresis type was represented by isolates of three distinct PspA clades. Sequence typing of pspA provides an added level of specificity in the subtyping of isolates and is a necessary first step in determining the components needed in a PspA vaccine which could elicit effective cross-protective coverage.
在一项近期针对1997年在美国不同地区分离出的对β-内酰胺耐药的肺炎球菌进行的基因分型调查中,8种克隆型(每种克隆型包含3至40株分离菌)占144株分离菌中的134株(G. 杰拉尔迪、C. 惠特尼、R. 法克拉姆和B. 比尔,《传染病杂志》181:216 - 229,2000年)。我们确定了这134株分离菌以及11株先前已鉴定的国际传播的多重耐药菌株中pspA基因的多样性程度。针对一个扩增子确定了34种不同的pspA限制性图谱,该扩增子包含编码PspA表面暴露结构域的结构基因可变部分以及一个可变长度的富含脯氨酸的假定细胞壁相关结构域。这些限制性图谱与对应于菌株Rx1 PspA第182至410位残基的约230个残基区域的33种不同pspA序列类型的图谱密切相关。这些残基包含一个100个残基的分支定义区域,已知该区域含有交叉保护性表位,共发现17种序列类型。在所评估的美国8种克隆型中的7种内,大多数菌株具有独特的、保守的pspA序列类型,而一种脉冲场凝胶电泳型由三个不同的PspA分支的分离菌代表。pspA序列分型在分离菌亚型分析中提供了更高的特异性水平,并且是确定PspA疫苗中能够引发有效交叉保护性覆盖所需成分的必要第一步。