本文關(guān)鍵詞： 豬鏈球菌 血清未分型 莢膜多糖基因簇 新cps型 Luminex wzy　出處：《中國疾病預防控制中心》2017年碩士論文　論文類型：學位論文

【摘要】：豬鏈球菌(Streptococcus sus)是一種重要的人獸共患病病原體。莢膜多糖(CPS)是豬鏈球菌的重要致病因子,在豬鏈球菌抗吞噬的過程中起關(guān)鍵作用,由基因組中的莢膜多糖基因簇(cps cluster)編碼。根據(jù)CPS抗原性不同,豬鏈球菌可分為33個血清型。凝集試驗和協(xié)同凝集試驗是豬鏈球菌傳統(tǒng)的血清分型方法。然而大量研究表明仍有一部分豬鏈球菌無法用傳統(tǒng)血清分型方法區(qū)分,即血清未分型菌株。在基因組時代,隨著高通量測序技術(shù)的普及,豬鏈球菌33種血清型標準菌株的基因組草圖完成測序。序列比對結(jié)果證實豬鏈球菌CPS的合成為Wzx/Wzy決定途徑,wzy基因是決定血清型別的關(guān)鍵基因。近年來,通過wzy基因的比較分析,11種新cps型別(NCL1～NCL10,Chz)的豬鏈球菌陸續(xù)被發(fā)現(xiàn)。在本研究的第一部分,我們對來自中國和加拿大的部分血清未分型豬鏈球菌進行了 Illumina高通量測序,并分析了其cps基因簇,由此發(fā)現(xiàn)了 10種新cps型別(NCL11～NCL20)�；谝郧暗难芯拷Y(jié)果,我們認為這些新cps型別極有可能代表新的豬鏈球菌血清型。除此之外,我們也對部分無法用傳統(tǒng)血清凝集方法區(qū)分,但經(jīng)分子血清分型方法鑒定為已知血清型的菌株進行了 Illumina高通量測序。通過對其cps基因簇的分析發(fā)現(xiàn)這些菌株的cps基因簇發(fā)生了不同程度的變異,例如基因的插入、缺失、替換,基因內(nèi)部的點突變以及cps基因簇的缺失等。我們的研究結(jié)果對揭示這部分菌株無法用抗血清分型的原因提供了線索。Luminex xMAP系統(tǒng)是基于多重微球的懸浮陣列平臺。32重Luminex體系已被建立用于同時檢測33種豬鏈球菌血清型。在本研究的第二部分,我們建立了一個可以同時檢測目前已發(fā)現(xiàn)的全部21種新cps型別豬鏈球菌的高通量檢測方法,即22重Luminex體系。該方法靈敏度高、特異性好,為檢測血清未分型豬鏈球菌提供了一種快速、有效、便捷的工具。
[Abstract]:Streptococcus sus is an important zoonotic pathogen. CPS is an important pathogenic factor of Streptococcus sus, which plays a key role in the anti-phagocytosis process of Streptococcus sus. Encoded by the capsule polysaccharide gene cluster in the genome, cps cluster.According to the antigenicity of CPS, Streptococcus suis can be divided into 33 serotypes. Agglutination test and co-agglutination test are the traditional serotyping methods of Streptococcus suis. In the genome era, with the popularity of high-throughput sequencing technology, The genomic sketches of 33 standard strains of Streptococcus suis serotype were sequenced. The results of sequence alignment confirmed that the synthesis of CPS from Streptococcus suis was the key gene to determine the Wzx/Wzy pathway. In the first part of this study, Illumina high-throughput sequencing of some untyped Streptococcus suis from China and Canada was carried out. The cps gene cluster was analyzed and 10 new cps types were found. Based on the previous research results, we think that these new cps types are very likely to represent new serotypes of Streptococcus suis. We also can't distinguish some of them with traditional seroagglutination. But the strains identified by molecular serotyping as known serotypes were sequenced by high throughput Illumina. By analyzing the cps gene cluster of these strains, it was found that the cps gene cluster of these strains had varying degrees of variation, such as gene insertion and deletion. Replace, Point mutation within the gene and deletion of cps gene cluster. Our results provide clues to the reason why these strains can not be typed with antiserum. Luminex xMAP system is based on multiple microspheres suspended array platform .32 heavy. Luminex system has been established for simultaneous detection of 33 serotypes of Streptococcus suis. We have established a high throughput detection method for all 21 new cps strains of Streptococcus suis, I. e., 22 Luminex system, which can simultaneously detect all the new strains of Streptococcus suis, which has high sensitivity and specificity. It provides a rapid, effective and convenient tool for the detection of untyped Streptococcus suis.
【學位授予單位】：中國疾病預防控制中心
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R378

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本文編號：1499906