基于液態(tài)懸浮芯片的鼠疫耶爾森氏菌SNP分型
發(fā)布時(shí)間：2018-11-17 06:44
【摘要】：鼠疫耶爾森氏菌(Yersinina pestis,以下簡稱鼠疫菌)曾引發(fā)世界范圍的三次鼠疫大爆發(fā),給人類帶來了災(zāi)難。鼠疫之所以能夠如此猖獗的肆掠整個(gè)世界,主要是因?yàn)槭笠咦鳛樽匀灰咴葱约膊∫云鋫鞑ニ俾蕵O快,突發(fā)病急不可預(yù)測,死亡率極高和傳染性極強(qiáng)為特征的甲類烈性傳染病。本研究利用多種遺傳標(biāo)記物對(duì)我國鼠疫東方型菌株進(jìn)行分型,可以避免采用單一遺傳標(biāo)記而導(dǎo)致的分辨率不足或偏性結(jié)果。主要研究結(jié)果如下： 1.建立了Luminex多重檢測SNP的技術(shù)平臺(tái)。鼠疫菌作為一種較為“年輕”且缺少基因重組的病原細(xì)菌,單核苷酸多態(tài)性(Single Nucleotide polymorphisms, SNP)可作為其非�？煽糠中秃拖到y(tǒng)發(fā)育分析指標(biāo)。本研究建立基于Lumienx x-TAG微球(液態(tài)懸浮芯片)的鼠疫菌SNP分型技術(shù),利用多重PCR同時(shí)擴(kuò)增鼠疫菌18個(gè)SNP位點(diǎn)片段,將擴(kuò)增完成的產(chǎn)物進(jìn)行多重ASPE (Allele Specific Primer Extension)反應(yīng)以及Luminex技術(shù)多重SNP檢測,通過一次反應(yīng)可以檢測出待測菌株中18個(gè)SNP位點(diǎn)的堿基特征。利用這一方法分析20株鼠疫東方型菌株,可以將其根據(jù)18個(gè)SNP位點(diǎn)的特征分為7個(gè)基因型,本方法具有判讀方便快速、高通量、成本低廉等特點(diǎn),在病原菌的SNP分型方面具有廣闊的應(yīng)用前景。 2.初步探討了我國鼠疫菌東方型的起源。目前國際上普遍認(rèn)為,鼠疫的第三次大流行起源于中國云南,鼠疫菌東方型菌株也起源于云南。但這與我們前期所研究的結(jié)果在某些方面與這一輪點(diǎn)不相符,即我們目前所測定的云南東方型菌株都是比較“年輕”的菌株,而中國最古老的東方型菌株分離自福建。本研究將運(yùn)用Luminex技術(shù)平臺(tái)檢測分析大量的云南東方型菌株,對(duì)云南東方型菌株有個(gè)初步的探討分型結(jié)果,我們對(duì)193株鼠疫東方型菌株進(jìn)行分型,一共分為13個(gè)基因型,分別是a, b, c, d, e, f, g, h, i, j, k,l, m。53個(gè)SNP位點(diǎn)信息對(duì)后期期追溯東方型鼠疫菌的起源提供了重要的數(shù)據(jù)。
[Abstract]:Yersinia pestis (Yersinina pestis, (Yersinia pestis) has caused three outbreaks of Yersinia pestis worldwide and brought disaster to human beings. The reason why plague is rampant throughout the world is mainly because plague as a natural epidemic disease is characterized by its rapid transmission rate, sudden and unpredictable disease, very high mortality and highly infectious type A strong infectious disease. In this study, a variety of genetic markers were used to type the oriental strains of plague in China, which could avoid the lack of resolution or bias caused by the use of single genetic marker. The main results are as follows: 1. The technical platform of Luminex multiple detection SNP is established. Yersinia pestis as a relatively "young" and lack of gene recombination of pathogenic bacteria, single nucleotide polymorphism (Single Nucleotide polymorphisms, SNP) can be used as a very reliable typing and phylogenetic analysis index. In this study, the SNP typing technique of Yersinia pestis based on Lumienx x-TAG microspheres (liquid suspension chip) was established, and 18 SNP loci of Yersinia pestis were amplified simultaneously by multiple PCR. The amplified products were detected by multiple ASPE (Allele Specific Primer Extension) reaction and Luminex technique by multiple SNP detection. The base characteristics of 18 SNP sites in the tested strain could be detected by one reaction. By using this method, 20 strains of Yersinia pestis oriental strain can be divided into 7 genotypes according to the characteristics of 18 SNP loci. The method is characterized by easy and fast interpretation, high throughput and low cost. It has a broad application prospect in SNP typing of pathogenic bacteria. 2. The oriental origin of Yersinia pestis in China was preliminarily discussed. At present, it is widely believed that the third pandemic of plague originated in Yunnan, China, and the oriental strain of Yersinia pestis also originated in Yunnan. However, this does not accord with the results of our previous studies in some respects, that is, the oriental strains in Yunnan are relatively "young" strains, and the oldest oriental strains in China are isolated from Fujian Province. In this study, a large number of oriental strains of Yunnan Province were detected and analyzed by using Luminex technology platform, and a preliminary study on the results of typing of oriental strains of Yunnan Province was carried out. We typed 193 strains of Oriental Yersinia pestis into 13 genotypes. The information of SNP loci of a, b, c, d, e, f, g, h, i, j, kanll and m.53 provides important data for tracing the oriental Yersinia pestis origin in late stage.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R378
本文編號(hào)：2336811
[Abstract]:Yersinia pestis (Yersinina pestis, (Yersinia pestis) has caused three outbreaks of Yersinia pestis worldwide and brought disaster to human beings. The reason why plague is rampant throughout the world is mainly because plague as a natural epidemic disease is characterized by its rapid transmission rate, sudden and unpredictable disease, very high mortality and highly infectious type A strong infectious disease. In this study, a variety of genetic markers were used to type the oriental strains of plague in China, which could avoid the lack of resolution or bias caused by the use of single genetic marker. The main results are as follows: 1. The technical platform of Luminex multiple detection SNP is established. Yersinia pestis as a relatively "young" and lack of gene recombination of pathogenic bacteria, single nucleotide polymorphism (Single Nucleotide polymorphisms, SNP) can be used as a very reliable typing and phylogenetic analysis index. In this study, the SNP typing technique of Yersinia pestis based on Lumienx x-TAG microspheres (liquid suspension chip) was established, and 18 SNP loci of Yersinia pestis were amplified simultaneously by multiple PCR. The amplified products were detected by multiple ASPE (Allele Specific Primer Extension) reaction and Luminex technique by multiple SNP detection. The base characteristics of 18 SNP sites in the tested strain could be detected by one reaction. By using this method, 20 strains of Yersinia pestis oriental strain can be divided into 7 genotypes according to the characteristics of 18 SNP loci. The method is characterized by easy and fast interpretation, high throughput and low cost. It has a broad application prospect in SNP typing of pathogenic bacteria. 2. The oriental origin of Yersinia pestis in China was preliminarily discussed. At present, it is widely believed that the third pandemic of plague originated in Yunnan, China, and the oriental strain of Yersinia pestis also originated in Yunnan. However, this does not accord with the results of our previous studies in some respects, that is, the oriental strains in Yunnan are relatively "young" strains, and the oldest oriental strains in China are isolated from Fujian Province. In this study, a large number of oriental strains of Yunnan Province were detected and analyzed by using Luminex technology platform, and a preliminary study on the results of typing of oriental strains of Yunnan Province was carried out. We typed 193 strains of Oriental Yersinia pestis into 13 genotypes. The information of SNP loci of a, b, c, d, e, f, g, h, i, j, kanll and m.53 provides important data for tracing the oriental Yersinia pestis origin in late stage.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R378
【參考文獻(xiàn)】
相關(guān)期刊論文 前6條
1 梁少生,林新勤,楊光華,秦石英,楊勤保,江超穗;一起大型水電站庫區(qū)鼠疫暴發(fā)流行的調(diào)查分析[J];中國地方病防治雜志;2001年03期
2 陳志強(qiáng);;“查士丁尼瘟疫”考辯[J];世界歷史;2006年01期
3 周冬生,楊瑞馥;細(xì)菌基因組進(jìn)化的分子策略[J];微生物學(xué)雜志;2004年01期
4 俞東征,海榮,董興齊,李敏,夏連續(xù),史獻(xiàn)明,魏建春,崔百忠,王鵬,孫連芝,張志凱,胡源,張恩民;中國鼠疫菌種資源遺傳特征分析[J];中華流行病學(xué)雜志;2003年11期
5 海榮,俞東征,魏建春,夏連續(xù),史獻(xiàn)明,張志凱,張恩民;中國鼠疫耶爾森菌的分子生物學(xué)特征與遺傳學(xué)意義[J];中華流行病學(xué)雜志;2004年06期
6 周志遠(yuǎn);;鼠疫簡史[J];生命世界;2009年06期
，本文編號(hào)：2336811
本文鏈接：http://sikaile.net/xiyixuelunwen/2336811.html
最近更新
教材專著
