本文選題：結(jié)核分枝桿菌 + 北京家族株��； 參考：《復(fù)旦大學(xué)》2013年碩士論文

【摘要】：結(jié)核病是一種對(duì)人類健康和社會(huì)穩(wěn)定造成嚴(yán)重威脅的傳染病,中國(guó)是結(jié)核病高負(fù)擔(dān)國(guó)家之一,盡管近些年來結(jié)核患病率有所下降,但我國(guó)的結(jié)核病疾病負(fù)擔(dān)依然很嚴(yán)重。W-北京基因型結(jié)核分枝桿菌是一類由共同祖先進(jìn)化而來、具有相似遺傳背景的M.TB菌株,在世界各地均有不同水平的流行。W-北京家族菌株可能與結(jié)核耐藥有關(guān),能夠引起暴發(fā),形成流行優(yōu)勢(shì)。 追蹤傳染源、闡明傳播途徑是結(jié)核病流行病學(xué)研究的兩大問題,結(jié)核具有潛伏感染的特點(diǎn),傳統(tǒng)的流行病學(xué)方法并不能很好地闡述其傳播機(jī)制,而以基因型分型技術(shù)為基礎(chǔ)的結(jié)核分子流行病學(xué)在這方面有不可替代的優(yōu)勢(shì)。如與傳統(tǒng)流行病學(xué)相結(jié)合,分子流行病學(xué)能更準(zhǔn)確地預(yù)測(cè)疾病的流行、暴發(fā),掌握其流行特征和傳播模式。因此,本研究選取華東地區(qū)的阜寧縣(江蘇省)、德清縣(江蘇省)和鄞州區(qū)(浙江省)為研究現(xiàn)場(chǎng),采用橫斷面調(diào)查和分子分型技術(shù)相結(jié)合的方法,描述華東地區(qū)W-北京基因型結(jié)核分枝桿菌的流行狀況,分析影響W-北京株流行的社會(huì)人口學(xué)特征、臨床治療史、既往治療史及人口流動(dòng)等因素；確定菌株的VNTR-MIRU基因型,評(píng)估結(jié)核病的近期傳播比例,探討華東地區(qū)結(jié)核病的傳播模式,為控制結(jié)核在人群中的傳播提供線索；比較不同位點(diǎn)VNTR-MIRU的分型效果及應(yīng)用前景,為完善結(jié)核分枝桿菌基因信息數(shù)據(jù)庫(kù)提供資料。 主要研究方法和結(jié)果如下： 1、W-北京基因型結(jié)核分枝桿菌的流行及影響因素研究 研究收集三個(gè)現(xiàn)場(chǎng)2009年6月/7月-2010年11月/12月在各區(qū)縣疾控中心登記的痰培養(yǎng)陽(yáng)性結(jié)核患者基本資料,收集患者的痰標(biāo)本進(jìn)行菌株培養(yǎng)、提取DNA,采用橫斷面研究結(jié)合DTM-PCR分型的方法,識(shí)別W-北京家族菌株及現(xiàn)代型W-北京株。研究共納入346例結(jié)核患者的M.TB菌株,經(jīng)RD105分型,90%(311株)屬于北京家族,阜寧、德清和鄞州的W-北京株檢出比例分別為91%(J64株)、96%(23株)和87%(124株),差別沒有統(tǒng)計(jì)學(xué)意義。W-北京株經(jīng)RD181進(jìn)一步分型,分別獲得現(xiàn)代型W-北京株294株(77%)、古典型W-北京株90株(23%)。分析年齡、健康狀況(BMI)等社會(huì)人口學(xué)因素及抗結(jié)核治療史等臨床特征對(duì)W-北京家族株流行的影響,除鄞州地區(qū)男性比女性具有更高的感染風(fēng)險(xiǎn)(OR=3.415,95%CI:1.071-10.884)外,未發(fā)現(xiàn)能夠明顯影響W-北京株流行的因素。 2、結(jié)核分枝桿菌耐藥基因型與傳播關(guān)系 采用直接測(cè)序法檢測(cè)與異煙肼、利福平耐藥有關(guān)的基因katG、rpoB。研究發(fā)現(xiàn),309株獲得katG基因的測(cè)序結(jié)果的M.TB菌株中,24株(7.77%)在katG基因315位存在突變,突變形式為AGC-ACC,其中23株屬于W-北京家族菌株；有145株M.TB菌株獲得rpoB基因測(cè)序結(jié)果,有7株(4.83%)發(fā)生531位突變,突變形式均為TCG-TTG,其中有6株(85.71%)為W-北京家族株。katG和rpoB基因在人群中的突變率差別沒有統(tǒng)計(jì)學(xué)意義。僅有1株發(fā)生katG基因突變的M.TB菌株發(fā)生成簇,突變菌株的成簇比例為4%；發(fā)生rpoB基因突變的M.TB菌株的基因型均為“唯一”型,無菌株發(fā)生成簇。 3、W-北京基因型結(jié)核分枝桿菌的基因型成簇特征及影響因素分析 采用7位點(diǎn)VNTR-MIRU分型方法對(duì)成功獲得DNA的426株M.TB菌株進(jìn)行基因分型,識(shí)別了409種基因型,包括395種“唯一”基因型和14個(gè)簇(包括31株分離株),每個(gè)簇內(nèi)有2-3株菌株不等,成簇比例為7.28%,近期傳播比例為3.99%。阜寧有29株形成13個(gè)簇,成簇比例為15%；德清有2株形成1個(gè)簇,成簇比例為2%；鄞州無菌株成簇,均為“唯一型”；三地區(qū)成簇比例存在顯著差異(x2=30.435,P0.001)。成簇菌株均為W_北京家族菌株,現(xiàn)代北京株的成簇率為7%,古典北京株成簇率為10%,二者的差別沒有統(tǒng)計(jì)學(xué)意義,并且發(fā)現(xiàn)現(xiàn)代型與古典型菌株之間發(fā)生成簇。 分析成簇患者的社會(huì)人口學(xué)及臨床特征發(fā)現(xiàn),戶籍是影響M.TB菌株成簇的因素之一,流動(dòng)人口的成簇風(fēng)險(xiǎn)明顯低于本地人口(aOR=0.107,95%CI:0.013-0.856)；未發(fā)現(xiàn)BCG接種史、結(jié)核治療史及痰涂片結(jié)果對(duì)菌株成簇有影響。成簇患者中有6例為復(fù)治病人,均為其所在簇內(nèi)最早出現(xiàn)結(jié)核癥狀的患者。在德清的成簇患者中,發(fā)現(xiàn)本地人口與流動(dòng)人口之間存在近期傳播。簇內(nèi)各患者呈散在分布,相互間沒有發(fā)現(xiàn)流行病學(xué)關(guān)聯(lián)。 4、VNTR-MIRU分型方法比較 對(duì)阜寧地區(qū)199株M.TB菌株采用國(guó)際通用的15位點(diǎn)VNTR-MIRU進(jìn)行基因分型,比較15位點(diǎn)與7位點(diǎn)VNTR-MIRU的分型能力。7位點(diǎn)組合中,各位點(diǎn)分辨力較高,最高為VNTR3820(0.8597),最低為Mtub21(0.6050);而15位點(diǎn)組合中,各個(gè)VNTR-MIRU位點(diǎn)分辨力水平有較大差異,從0.0804(ETRC)到0.7753(MIRU26)不等。15位點(diǎn)組合對(duì)阜寧地區(qū)M.TB菌株及北京株的總分辨力(HGI值)均為0.9994,而7位點(diǎn)組合的分辨力分別為0.9990和0.9988,略低于15位點(diǎn)組合。MIRU-15將阜寧199株M.TB菌株分為190種基因型,16株菌株成7個(gè)簇,成簇比例為8%,顯著低于MIRU-7的成簇率(8%vs.15%,x2=4.234,P=0.040)。位點(diǎn)聯(lián)合使用可以提高VNTR-MIRU分型方法的分辨力,但過多增加位點(diǎn)數(shù)量并不能使總分辨力得到進(jìn)一步提高。 通過本研究,我們可以得出如下結(jié)論： 1.W-北京家族菌株在華東地區(qū)的流行水平很高(90%),且沒有發(fā)現(xiàn)年齡、健康狀況(BMI)、戶籍等社會(huì)人口學(xué)因素與北京株的分布的關(guān)聯(lián),也沒有發(fā)現(xiàn)BCG接種史、結(jié)核治療史、痰涂片結(jié)果等臨床特征與北京株的流行有關(guān)。 2.W-北京株中,現(xiàn)代北京株的比例(77%)高于古典北京株,且現(xiàn)代北京株的分布不因社會(huì)人口學(xué)和臨床特征而不同,始終占有較高的流行比例,提示現(xiàn)代型可能具有較強(qiáng)的致病性。 3.研究地區(qū)M.TB菌株成簇比例較低,提示華東地區(qū)結(jié)核病的發(fā)生主要是由既往感染結(jié)核復(fù)發(fā)造成,存在較小比例的近期傳播,而且未發(fā)現(xiàn)簇內(nèi)患者之間存在流行病學(xué)關(guān)聯(lián),提示由偶然接觸導(dǎo)致結(jié)核發(fā)生可能是華東地區(qū)結(jié)核病傳播的主要模式。 4.戶籍是影響M.TB菌株成簇的主要因素之一,本地人口的成簇風(fēng)險(xiǎn)顯著高于流動(dòng)人口,提示本地人口比流動(dòng)人口更容易發(fā)生結(jié)核的近期傳播,同時(shí)研究也發(fā)現(xiàn)本地人口與流動(dòng)人口之間能發(fā)生結(jié)核的交叉?zhèn)鞑ァ?5.推薦選擇來自MIRU-7的VNTR3820、Qub11a、Qub18和來自MIRU-15的Qub11b、MIRU10、MIRU26、MIRU31位點(diǎn)進(jìn)行華東地區(qū)M.TB菌株的基因分型研究。
[Abstract]:Tuberculosis is an infectious disease that poses a serious threat to human health and social stability. China is one of the countries with a high burden of tuberculosis. Despite the decline in the prevalence of tuberculosis in recent years, the burden of tuberculosis disease in China is still very serious..W- Beijing Mycobacterium tuberculosis is a kind of common ancestor evolved from a common ancestor. M.TB strains of genetic background, in all parts of the world, have different levels of epidemic.W- Beijing family strains, which may be related to tuberculosis resistance, which can cause outbreak and form epidemic advantages.
Tracing the source of infection and clarifying the way of transmission is the two major problem in the epidemiological study of tuberculosis. Tuberculosis has the characteristics of latent infection. The traditional epidemiological method can not explain its transmission mechanism well, and the molecular epidemiology of tuberculosis based on genotyping technology has an irreplaceable advantage in this respect. With the combination of diseases and epidemiology, molecular epidemiology can predict the epidemic and outbreak of disease more accurately, and grasp its epidemic characteristics and mode of transmission. Therefore, this study selects Funing county (Jiangsu province), Deqing County (Jiangsu province) and Yinzhou District (Zhejiang province) in East China as the research site, and uses a method of combining cross-sectional investigation and molecular typing to describe the method of combining cross-sectional investigation and molecular typing. The epidemic status of Mycobacterium tuberculosis in W- Beijing genotypes in East China was analyzed. The social demographic characteristics, history of clinical treatment, history of treatment and population flow and other factors affecting the epidemic of W- strains were analyzed. The VNTR-MIRU genotype of the strain was determined, the rate of tuberculosis was assessed in the near future, and the mode of tuberculosis transmission in East China was discussed. It provides clues for the spread of tuberculosis in the population, compares the typing effect of VNTR-MIRU with different loci and its application prospects, and provides information for improving the genetic information database of Mycobacterium tuberculosis.
The main research methods and results are as follows:
1, the prevalence and influencing factors of W- Mycobacterium tuberculosis in Beijing
The basic data of sputum positive tuberculosis patients registered in each district and county CDC in June 2009 /7 month -2010 and /12 month in November /12 month were collected and collected. The sputum specimens of the patients were collected for strain culture, and DNA was extracted. A cross-sectional study combined with DTM-PCR typing was used to identify the W- Beijing family strain and the modern W- Beijing strain. M.TB strains of 346 tuberculosis patients, RD105 classification, 90% (311 strains) belong to Beijing family, Funing, Deqing and Yinzhou W- Beijing strains were 91% (J64 strain), 96% (23 strains) and 87% (124 strains), the difference was not statistically significant.W- Beijing strains through the one step type, respectively, the modern W- Beijing strain 294 strains (77%), the classical W- Beijing, respectively. Strain 90 (23%). Analysis of the social demographic factors such as age, health status (BMI) and the history of anti tuberculosis treatment history and other clinical characteristics on the epidemic of W- Beijing family, except for the higher risk of infection (OR=3.415,95%CI:1.071-10.884) of men in Yinzhou than women (OR=3.415,95%CI:1.071-10.884), there is no obvious influence on the factors of the epidemic of the W- Beijing strain.
2, the relationship between drug-resistant genotypes and transmission of Mycobacterium tuberculosis
The direct sequencing method was used to detect the gene katG related to isoniazid and rifampicin resistance. RpoB. studies found that 24 (7.77%) of the M.TB strains obtained from the sequencing of katG gene were mutated in the katG gene, and the mutation was AGC-ACC, 23 of which belonged to the W- Beijing family strain; 145 M.TB strains obtained the rpoB gene sequencing node. 531 mutations were found in 7 (4.83%) strains, all of which were TCG-TTG, of which 6 (85.71%) were W- Beijing family.KatG and rpoB gene mutations in the population with no statistical significance. Only 1 strains of katG gene mutation were produced by M.TB strains, and the proportion of mutant strains was 4%; rpoB gene mutation of M.TB bacteria. The genotypes of the plants were all "only", and the aseptic plants were clustered.
3, genotypic clustering characteristics and influencing factors of W- Beijing Mycobacterium tuberculosis
The 7 loci VNTR-MIRU typing method was used to genotyping 426 strains of M.TB strains, which were successfully obtained. 409 genotypes were identified, including 395 "unique" genotypes and 14 clusters (including 31 isolates). There were 2-3 strains in each cluster, the cluster ratio was 7.28%, the proportion of the near phase transmission was 29 in 3.99%. Funing, and 13 clusters were formed. The proportion of Yinzhou was 15%, 1 clusters were formed in Deqing and 2% in cluster proportion; all the sterile plants in Yinzhou were "unique". There were significant differences in the cluster proportion of three regions (P0.001). The cluster strains were all W_ Beijing family strains, the clustering rate of modern Beijing strain was 7%, and the clustering rate of Beijing strain in ancient Scripture was 10%, and there was no statistical difference between two. Significance and discovery of clusters between modern and classical strains.
The social demography and clinical characteristics of the cluster patients showed that the household registration was one of the factors affecting the cluster of M.TB strains. The cluster risk of the floating population was significantly lower than that of the local population (aOR=0.107,95%CI:0.013-0.856); the history of BCG inoculation, the history of tuberculosis treatment and the sputum smear results affected the strain of the strain. 6 of the cluster patients were retreated. Patients were the earliest patients with tuberculosis symptoms in their cluster. In the cluster patients of Deqing, there was a recent spread between the local population and the floating population. The patients were scattered in clusters, and there was no epidemiological association between each other.
4, comparison of VNTR-MIRU typing methods
In Funing, 199 strains of M.TB strains were genotyping by international VNTR-MIRU loci VNTR-MIRU. In the combination of the 15 loci and the 7 locus VNTR-MIRU, the resolution of each point was higher, the highest was VNTR3820 (0.8597) and the lowest was Mtub21 (0.6050), while the resolution level of the VNTR-MIRU loci in the 15 bit combination had a great difference. The total resolution of M.TB strain and Beijing strain in Funing area from 0.0804 (ETRC) to 0.7753 (MIRU26) is 0.9994, and the resolution of the 7 loci combination is 0.9990 and 0.9988 respectively, and the 7 loci combination.MIRU-15 divides 199 strains of M.TB strains in Funing into 190 genotypes, 16 strains are divided into 7 clusters, and the cluster proportion is 8. %, significantly lower than the clustering rate of MIRU-7 (8%vs.15%, x2=4.234, P=0.040). The combined use of loci can improve the resolution of the VNTR-MIRU typing method, but the number of excessive increasing loci can not further improve the total resolution.
Through this study, we can draw the following conclusions:
The prevalence of 1.W- Beijing family strains in East China was very high (90%), and no age, health status (BMI), household registration and other social demographic factors were associated with the distribution of Beijing strains. There was no history of BCG inoculation, tuberculosis treatment history, sputum smear results and other clinical features related to the epidemic of Beijing strain.
In the 2.W- Beijing strain, the proportion of modern Beijing strains (77%) is higher than that of the classical Beijing strain, and the distribution of modern Beijing strains is not different from the social demography and clinical characteristics, and it always occupies a high prevalence rate, suggesting that modern type may have a strong pathogenicity.
3. the cluster proportion of M.TB strains in the study area is low, suggesting that the occurrence of tuberculosis in East China is mainly caused by the recurrence of tuberculosis, there is a small proportion of the recent transmission, and there is no epidemiological association between the patients in the cluster. It is suggested that the occurrence of tuberculosis from accidental contact may be the main transmission of tuberculosis in East China. Pattern.
The 4. domicile is one of the main factors affecting the cluster of M.TB strains. The local population has a higher risk of clustering than the floating population, suggesting that the local population is more likely to have the recent spread of tuberculosis than the floating population, and the study also finds that the local population and the floating population can cross the cross transmission of tuberculosis.
5. recommend VNTR3820, Qub11a, Qub18 and Qub11b, MIRU10, MIRU26, and MIRU31 from MIRU-7, MIRU10, MIRU26, and MIRU31 loci to study the genotyping of M.TB strains in East China.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R52;R181.3

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4 張子勝;曹光農(nóng);;福建省結(jié)核病控制督導(dǎo)工作模式探討[A];中國(guó)防癆雜志2003第25卷增刊——2003年中國(guó)防癆協(xié)會(huì)全國(guó)學(xué)術(shù)會(huì)議論文集[C];2003年

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6 方小平;趙玉杭;陳韶華;朱國(guó)雄;;福州市結(jié)核病防治工作可持續(xù)發(fā)展的研究[A];中國(guó)防癆雜志2003第25卷增刊——2003年中國(guó)防癆協(xié)會(huì)全國(guó)學(xué)術(shù)會(huì)議論文集[C];2003年

7 胡達(dá)敏;王燁源;;蘇州市結(jié)核病控制規(guī)劃的實(shí)施與效果[A];中國(guó)防癆雜志2003第25卷增刊——2003年中國(guó)防癆協(xié)會(huì)全國(guó)學(xué)術(shù)會(huì)議論文集[C];2003年

8 吳浩;王年保;李麗娟;;城鄉(xiāng)結(jié)合部流動(dòng)人口結(jié)核病控制[A];中國(guó)防癆雜志2003第25卷增刊——2003年中國(guó)防癆協(xié)會(huì)全國(guó)學(xué)術(shù)會(huì)議論文集[C];2003年

9 肖東樓;;結(jié)核病防治工作的戰(zhàn)略機(jī)遇與可持續(xù)發(fā)展[A];中華醫(yī)學(xué)會(huì)結(jié)核病學(xué)分會(huì)2004年學(xué)術(shù)會(huì)議論文匯編[C];2004年

10 方小平;陳韶華;趙玉杭;朱國(guó)雄;;福州市開展結(jié)核病控制可持續(xù)發(fā)展的實(shí)施性研究(摘要)[A];中華醫(yī)學(xué)會(huì)結(jié)核病學(xué)分會(huì)2004年學(xué)術(shù)會(huì)議論文匯編[C];2004年

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本文編號(hào)：2009758