A molecular-capsular-type prediction system for 90 Streptococcus pneumoniae serotypes using partial cpsA-cpsB sequencing and wzy- or wzx-specific PCR
A molecular-capsular-type prediction system for 90 Streptococcus pneumoniae serotypes using partial cpsA-cpsB sequencing and wzy- or wzx-specific PCR
复制标题
DOI：
10.1099/jmm.0.45924-0
复制
发表时间：
2005-04-01
影响因子：
3
通讯作者：
Gilbert, GL
中科院分区：
文献类型：
--
作者：
Kong, FR;Wang, WZ;Gilbert, GL
在先前的研究中，基于cPSA-cpsB部分测序和S(S)/S(S)血清型特异性聚合酶链式反应，建立了51个肺炎链球菌血清型的分子包膜型预测系统。在这项研究中，另外169株肺炎链球菌被添加到现有的427株分离株的数据库中，包括所有39个以前没有研究过的血清型的代表。除了作者自己的90个血清型的有限序列数据外，肺炎链球菌衣壳基因座测序小组在桑格研究所发表的或可从GenBank获得的cPSA-cpsB序列数据也被纳入数据库。除25A外，所有血清型均由至少两个分离物代表。鉴定的序列类型有138个，其中110个对应于单一的常规血清型；其中57个由两个或两个以上的分离物代表。26种序列类型由2到4个CS共享。为了解析这些共有的cPSA-cpsB序列类型，提高我们系统的识别力，我们对编码衣壳多糖翻转酶(Wzx)和聚合酶(Wzy)的基因进行了注释，并设计了24个针对wzy的新的血清型(S)/群(S)特异的PCR，以及两个针对wzx的新的针对wzx的PCR。使用cPSA-cpsB测序和wzx/wzy PCR，MCT正确地预测了516株(73%)的css和708株被评估的另外155株(22%)的血清群。对于5%的分离物，MCT不能区分包含不同血清组成员的五个血清型对(37个分离物)的成员。虽然MCT和CS之间的关系还需要进一步研究，但该系统现在可以对95%的肺炎链球菌分离株进行血清型或血清型鉴定。
In a previous study, a molecular capsular type (MCT) prediction system for 51 Streptococcus pneumoniae serotypes was developed based on a combination of partial cpsA-cpsB sequencing and serotype(s)/group(s)-specific PCR. In this study, another 169 S. pneumoniae isolates were added to the existing database of 427 isolates, including representatives of all 39 serotypes not previously studied. In addition to the authors' own limited sequence data for ail 90 serotypes, cpsA-cpsB sequence data published by the S. pneumoniae capsular loci-sequencing group (http:// www.sanger.ac.uk/Projects/S_pneumoniae/CPS/) at the Sanger Institute or available from GenBank were incorporated into the database. All serotypes, except 25A, were represented by at least two isolates. The number of sequence types identified was 138, of which 110 corresponded to single conventional serotypes (CSs); of these, 57 were represented by two or more isolates. Twenty-six sequence types were shared by between two and four CSs. To resolve these shared cpsA-cpsB sequence types and increase the discriminatory power of our system, the genes encoding the capsular polysaccharide flippase (wzx) and polymerase (wzy) were annotated and 24 new serotype(s)/group(s)-specific PCRs targeting wzy and two targeting wzx were designed. Using both cpsA-cpsB sequencing and wzx/wzy PCR, MCT correctly predicted the CSs of 516 (73 %) and the serogroup of an additional 155 (22 %) of the 708 isolates evaluated. For 5 % of isolates, MCT could not distinguish between members of five serotype pairs (37 isolates) containing members of different serogroups. Although further study of the relationship between MCT and CS is needed, this system now allows serotype or serogroup identification of 95 % of S. pneumoniae isolates.