【摘要】： 本課題以霍亂弧菌特異性的霍亂腸毒素A亞單位基因(ctxA)、毒素協(xié)同調(diào)節(jié)菌毛A基因(tcpA)和全DNA序列為目標(biāo),通過聚合酶鏈反應(yīng)(PCR)技術(shù)、限制性內(nèi)切酶技術(shù)、脈沖場凝膠電泳技術(shù)和BioNumerics V4.0軟件UPGMA方法進(jìn)行聚類分析,觀察了來源于重慶市1992年至2006年的疫情和環(huán)境檢測樣本的94株霍亂弧菌核酸分子特征。 著重探索霍亂暴發(fā)疫情分離的菌株與菌株之間,疫情分離的菌株與海、水產(chǎn)品監(jiān)測分離的霍亂弧菌之間的遺傳相關(guān)性,推斷疫情菌株的來源、變遷及分子指紋圖譜變化規(guī)律,以期為預(yù)測疫情和制定防治措施提供依據(jù)。 研究結(jié)果顯示: 1、94株霍亂弧菌樣本(83株感染者樣本,11株環(huán)境樣本)中ctxA基因陽性檢出92株(97.87%),2株水產(chǎn)品市場監(jiān)測分離菌株為陰性;tcpA基因陽性檢出91株(96.81%),2株水產(chǎn)品市場監(jiān)測分離菌株和1株來源于病人的分離株為陰性。 2、94株霍亂弧菌被分為44種脈沖場凝膠電泳圖譜,聚類分析將大部分菌株分為三類(小川型,稻葉型,O139群)。 3、2002年重慶市部分區(qū)縣多點暴發(fā)霍亂疫情埃爾托稻葉型1d霍亂弧菌菌株基因同源性較高;1992-2006年重慶市埃爾托小川型和埃爾托稻葉型相似系數(shù)僅為75%;從甲魚分離的O139群霍亂弧菌與同期流行的O139群霍亂弧菌優(yōu)勢菌株的脈沖場凝膠電泳圖譜型別一致,并且也為產(chǎn)毒株。 由此可見,1992-2006年重慶市霍亂弧菌流行株存在緊密相關(guān)的核酸分子特征;經(jīng)過長期的霍亂流行后埃爾托霍亂弧菌主要向小川型和稻葉型分化,兩型分子特征差異較大;甲魚中分離的O139群霍亂弧菌與霍亂疫情分離菌株之間高度同源,被污染的甲魚可能是近年來食源性霍亂暴發(fā)的主要傳染來源之一,提示:海、水產(chǎn)品的監(jiān)測應(yīng)當(dāng)是霍亂防控的重點。
[Abstract]:The aim of this study was to co-regulate the (tcpA) and DNA sequences of the pili A gene of Vibrio cholerae specific enterotoxin A subunit gene (ctxA), toxin, and to use polymerase chain reaction (PCR) technique and restriction endonuclease technique. The nucleic acid molecular characteristics of 94 strains of Vibrio cholerae from 1992 to 2006 in Chongqing were observed by cluster analysis with pulsed field gel electrophoresis (PGE) technique and UPGMA method of BioNumerics V4.0 software. The genetic correlation between strains isolated from cholera outbreak and strains, between strains isolated from epidemic situation and sea, and between Vibrio cholerae isolated by aquatic product monitoring was explored, and the source, changes and variation rules of molecular fingerprints of the strains were inferred. In order to predict the epidemic situation and formulate prevention and control measures to provide the basis. The results showed that: 1 92 strains (97.87%) were positive for ctxA gene in 94 strains of Vibrio cholerae (83 infected strains, 11 environmental samples), and 2 strains were negative in aquatic products market monitoring. 91 strains (96.81%) were positive for tcpA gene, 2 strains were negative for aquatic products market monitoring and 1 strain was isolated from patients. Two hundred and ninety-four strains of Vibrio cholerae were divided into 44 kinds of pulsed field gel electrophoresis patterns, and most of them were classified into three groups (Ogawa type, rice leaf type, O139 group) by cluster analysis. (3) in 2002, there was a high homology among vibrio cholerae strains of Eltorice leaf type 1d in some districts and counties of Chongqing in 2002, and the similarity coefficient of leaf type between Erto Xiaochuan type and Elto leaf type in Chongqing from 1992 to 2006 was only 75%. The dominant strains of Vibrio cholerae O139 isolated from Turtle were identical to the dominant strains of Vibrio cholerae O139 in the same period, and were also virulent strains. It can be seen that the epidemic strains of Vibrio cholerae in Chongqing from 1992 to 2006 had closely related nucleic acid molecular characteristics, and after a long period of cholera epidemic, Vibrio cholerae mainly differentiated into Ogawa type and rice leaf type, and the molecular characteristics of the two types were quite different. The isolates of Vibrio cholerae O139 isolated from turtle have high homology with the isolates of cholera epidemic situation. The contaminated turtle may be one of the main infectious sources of foodborne cholera outbreaks in recent years. Surveillance of aquatic products should be the focus of cholera prevention and control.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R378

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 闞飆;;霍亂監(jiān)測的新問題[J];傳染病信息;2006年01期

2 肖邦忠,羅興建,吳國輝,李秀安,張靜秋,龍江,戚小東;一起食源性霍亂暴發(fā)的調(diào)查分析[J];第三軍醫(yī)大學(xué)學(xué)報;2003年11期

3 劉延清;霍亂的分子流行病學(xué)[J];海峽預(yù)防醫(yī)學(xué)雜志;2002年01期

4 張政,劉彩蓮,闞飆,劉延清;霍亂弧菌16SrRNA基因分型研究[J];疾病監(jiān)測;2000年09期

5 闞飆,徐建國;傳染病監(jiān)測的實驗室網(wǎng)絡(luò)化[J];疾病監(jiān)測;2005年01期

6 扈慶華,鄭薇薇,石曉路,葉寶英,李慶閣,王冰,莊志雄,劉小立;改良分子信標(biāo)—實時PCR快速檢測霍亂弧菌[J];中國熱帶醫(yī)學(xué);2004年04期

7 肖邦忠,羅興建,吳國輝,龍江,張靜秋,肖達(dá)勇;13起O139型霍亂疫情分析[J];熱帶醫(yī)學(xué)雜志;2005年03期

8 羅興建;肖邦忠;張靜秋;吳國輝;易娟;龍江;;重慶市2002年霍亂流行特征分析[J];熱帶醫(yī)學(xué)雜志;2006年05期

9 周曉明,盧林耿,衛(wèi)國榮,張幼辰,俞順章;脈沖場電泳進(jìn)行E.coliO_(157)的分型[J];復(fù)旦學(xué)報(醫(yī)學(xué)科學(xué)版);2001年01期

10 芮勇宇,蔡初的,蕭斌權(quán),俞守義,李建基,廖育煌,王紅,鐘豪杰,羅不凡;隨機(jī)引物PCR方法用于霍亂分子流行病學(xué)研究[J];中國公共衛(wèi)生;2000年07期

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