河西走廊地域流行幽門螺桿菌菌庫(kù)建立、耐藥狀況及重要毒力基因分型研究
發(fā)布時(shí)間:2019-06-20 23:03
【摘要】:目的: 1.從河西走廊地域上消化道疾病患者體內(nèi)分離鑒定幽門螺桿菌(Helicobacter Pylori, Hp),建立胃癌高發(fā)區(qū)區(qū)域性Hp菌庫(kù)。為進(jìn)一步研究Hp生物學(xué)特性、致病性和致病機(jī)制、研制有效疫苗打下基礎(chǔ)。 2.調(diào)查河西走廊地域流行Hp菌株對(duì)常用抗菌藥物(阿莫西林、克林霉素、左氧氟沙星、甲硝唑、呋哺唑酮、四環(huán)素)的耐藥狀況,為當(dāng)?shù)豀p根除提供依據(jù)。 3.調(diào)查河西走廊地域流行Hp重要毒力基因vacA和cagA基因型,分析不同基因型Hp感染與河西走廊上消化道疾病發(fā)生的關(guān)系。方法: 1.收集武威、金昌、張掖三地醫(yī)院314例上消化道疾病患者胃粘膜標(biāo)本,經(jīng)微需氧分離培養(yǎng),觀察培養(yǎng)特性、形態(tài)學(xué)、快速尿素酶、氧化酶、觸酶及ureh.PCR擴(kuò)增,鑒定Hp。收集相關(guān)信息,統(tǒng)一編號(hào),-80℃凍存,建立河西走廊地域流行Hp菌庫(kù)。 2.應(yīng)用Hp菌庫(kù),采用Kirby-Bauer紙片法檢測(cè)71株河西走廊地域流行Hp菌株對(duì)阿莫西林、克拉霉素、左氧氟沙星、甲硝唑、呋哺唑酮、四環(huán)素的耐藥狀況。比較不同性別、年齡段、病理類型患者Hp菌株的耐藥差異。 3.應(yīng)用Hp菌庫(kù),提取70株Hp基因組DNA。 PCR擴(kuò)增Hp vacA基因s區(qū)(sla,slb,slc,s2)和m區(qū)(ml和m2)以及cagA。分析不同的vacA和cagA基因型與臨床病理特征、年齡、性別以及地域的關(guān)系。結(jié)果: 1.從314例上消化道疾病患者中分離到Hp81株,陽(yáng)性率為25.80%。不同性別患者間Hp陽(yáng)性率差異無統(tǒng)計(jì)學(xué)意義(χ2=0.427,P0.05)。不同病理類型患者間Hp陽(yáng)性率差異有統(tǒng)計(jì)學(xué)意義(χ=7.705,P0.05)。其中消化性潰瘍組(46.43%)顯著高于慢性胃炎組(23.32%)(P0.05)。不同年齡段患者間Hp陽(yáng)性率差異有統(tǒng)計(jì)學(xué)意義(χ2=13.974,P0.05),其中15-30歲年齡組人群Hp分離陽(yáng)性率顯著高于61歲以上年齡組。液氮保存組胃黏膜標(biāo)本Hp分離陽(yáng)性率顯著高于-20℃保存組(χ2=5.368,P0.05)。-80℃凍存菌株,成功建立河西走廊地域流行Hp菌庫(kù)。 2.71株Hp對(duì)常用6種抗菌藥物的耐藥性依次為:甲硝唑(47.89%),左氧氟沙星(8.45%),阿莫西林(4.23%),四環(huán)素(2.82%),克拉霉素(1.41%),呋哺唑酮(0%)。Hp對(duì)不同抗菌藥物的耐藥率在不同性別、不同年齡段間均無顯著性差異(P0.05)。分離自慢性胃炎、消化性潰瘍、胃癌患者菌株對(duì)左氧氟沙星的耐藥率分別為3.85%(2/52)、9.09%(1/11)和37.50%(3/8),差異有統(tǒng)計(jì)學(xué)意義(P0.05),其中胃癌組分離Hp對(duì)左氧氟沙星的耐藥率高于慢性胃炎組(P0.05),對(duì)其它抗菌藥物的耐藥狀況均無顯著性差異(P0.05)。 3.70株Hp檢出vacA陽(yáng)性率為100%, cagA陽(yáng)性率為97.14%(68/70)。sla單陽(yáng)性菌株40株,陽(yáng)性率57.14%(40/70);s1c單陽(yáng)性菌株11株,陽(yáng)性率15.71%(11/70),sla和s1c陽(yáng)性混合感染菌株19株,陽(yáng)性率27.14%(19/70)。檢出m2陽(yáng)性菌株37株,陽(yáng)性率為52.86%,未檢出ml亞型。vacAsla型以及sla/m2型Hp在不同病理類型、年齡段和地區(qū)患者中的感染率高于其它基因型Hp。結(jié)論: 1.Hp感染在河西走廊地域消化性潰瘍的發(fā)生和發(fā)展中起重要作用。 2.在轉(zhuǎn)運(yùn)培養(yǎng)基中加入甘油,通過液氮保存和運(yùn)送可提高遠(yuǎn)距離、采集后放置時(shí)間較長(zhǎng)胃粘膜標(biāo)本中Hp的分離率。 3.河西地域流行Hp菌庫(kù)的建立,將為進(jìn)一步研究Hp致病機(jī)制、遺傳進(jìn)化、疫苗研制等打下基礎(chǔ)。 4.河西走廊地域流行Hp對(duì)甲硝唑耐藥率較高,在一線根除方案中應(yīng)盡量避免使用;呋哺唑酮、阿莫西林等應(yīng)作為根除的主要藥物。 5.河西走廊地域流行菌株主要為vacA和cagA雙陽(yáng)性,致病性強(qiáng)的Ⅰ型菌株;這可能是河西走廊地域消化性潰瘍、胃癌高發(fā)的原因之一。流行菌株中vacA基因型以sla/m2為主,其次為slc/m2。 vac Am區(qū)可能存在較大變異。
[Abstract]:Purpose: 1. Isolation and identification of Helicobacter pylori (Hp) from patients with upper gastrointestinal diseases in Hexi Corridor, and to establish regional Hp (Hp) bacteria in high-hair region of gastric cancer Library. To further study the biological characteristics, pathogenicity and pathogenesis of Hp, and to develop an effective vaccine. 2. To investigate the drug-resistant status of the Hp strains in the regional epidemic of Hexi Corridor to the common antibacterial drugs (amoxicillin, clindamycin, levofloxacin, sodium nitrogenide, and chlortetracycline, tetracycline), and to eliminate the Hp eradication in the local area. Objective:3. To investigate the major virulence gene vacA and cagA genotype in the region of Hexi Corridor, and to analyze the different genotypes of Hp infection and upper gastrointestinal diseases in the Hexi Corridor. occurring Methods:1. The gastric mucosa specimens of 314 patients with upper digestive tract diseases were collected from the three hospitals in Wuwei, Jinchang and Zhangye. The culture characteristics, morphology, rapid urease, oxidase, enzyme and ureh were observed by microaerobic separation culture. Information, uniform number,-80 鈩,
本文編號(hào):2503584
[Abstract]:Purpose: 1. Isolation and identification of Helicobacter pylori (Hp) from patients with upper gastrointestinal diseases in Hexi Corridor, and to establish regional Hp (Hp) bacteria in high-hair region of gastric cancer Library. To further study the biological characteristics, pathogenicity and pathogenesis of Hp, and to develop an effective vaccine. 2. To investigate the drug-resistant status of the Hp strains in the regional epidemic of Hexi Corridor to the common antibacterial drugs (amoxicillin, clindamycin, levofloxacin, sodium nitrogenide, and chlortetracycline, tetracycline), and to eliminate the Hp eradication in the local area. Objective:3. To investigate the major virulence gene vacA and cagA genotype in the region of Hexi Corridor, and to analyze the different genotypes of Hp infection and upper gastrointestinal diseases in the Hexi Corridor. occurring Methods:1. The gastric mucosa specimens of 314 patients with upper digestive tract diseases were collected from the three hospitals in Wuwei, Jinchang and Zhangye. The culture characteristics, morphology, rapid urease, oxidase, enzyme and ureh were observed by microaerobic separation culture. Information, uniform number,-80 鈩,
本文編號(hào):2503584
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