發(fā)布時(shí)間：2018-05-26 14:07

  本文選題：人芽囊原蟲(chóng) + 感染情況��； 參考：《廣西醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的 了解廣西兩所三甲醫(yī)院住院患者人芽囊原蟲(chóng)(Blastocystis hominis,B.h)的感染情況；確定住院患者B.h感染株的基因分型,揭示未知基因型B.h分離株的遺傳距離及遺傳多態(tài)性；探討甲硝唑、替硝唑、阿奇霉素、諾氟沙星4種藥物對(duì)B.h的作用效果。方法收取廣西三甲醫(yī)院(廣西醫(yī)科大學(xué)第一附屬醫(yī)院、廣西醫(yī)科大學(xué)附屬腫瘤醫(yī)院,以下分別簡(jiǎn)稱一附院和腫瘤醫(yī)院)住院患者新鮮糞便,采用改良酸醚沉淀法,鏡檢定性診斷,用R語(yǔ)言軟件分析住院患者與正常人群、胃腸疾病患者與非胃腸疾病患者、兩所醫(yī)院住院患者性別、職業(yè)、年齡、民族及受教育水平各組間B.h感染率。收集經(jīng)鑒定的陽(yáng)性蟲(chóng)體,37℃厭氧培養(yǎng),提取陽(yáng)性蟲(chóng)體DNA；以基因組DNA為模板,采用7對(duì)STS特異性引物進(jìn)行PCR擴(kuò)增明確其基因分型；把未知基因型分離株作為研究對(duì)象應(yīng)用RAPD標(biāo)記技術(shù)構(gòu)建條帶圖譜,用NTSYSpc軟件對(duì)分離株的遺傳多樣性進(jìn)行聚類(lèi)分析,構(gòu)建聚類(lèi)分析圖。最后采用MTT法檢測(cè)B.h對(duì)甲硝唑、替硝唑、阿奇霉素、諾氟沙星的藥敏反應(yīng),觀察4種藥物對(duì)B.h的作用效果。結(jié)果(1)兩所醫(yī)院共檢查住院患者2800例,總陽(yáng)性率為36.6%。附院與腫瘤醫(yī)院陽(yáng)性率分別為22.59%、43.24%。與正常人群感染率相比,住院患者感染率高于正常人群(P=0.000)；腫瘤患者與非腫瘤患者、正常人群感染率相比,差異有統(tǒng)計(jì)學(xué)意義(P=0.000)；按照胃腸科室與非胃腸科室分組,兩組感染率差異有統(tǒng)計(jì)學(xué)意義(P=0.000)；按照文化程度、職業(yè)劃分,兩所醫(yī)院初中及以下學(xué)歷患者(P=0.000)、農(nóng)民患者感染率較其他文化程度患者感染率差異有統(tǒng)計(jì)學(xué)意義(P=0.000)；按照性別、年齡、民族分組,兩所醫(yī)院各組間感染率差異沒(méi)有統(tǒng)計(jì)學(xué)意義(P0.05)。(2)STS基因分型結(jié)果顯示,共57例標(biāo)本,有36例標(biāo)本被分型,其中單純1型2株、單純2型20株、單純3型3株、單純4型3株、單純6型5株、單純7型3株,剩余21例為未知基因型；1型、4型、7型共擴(kuò)增出8株,且8株均為一附院住院患者分離株,3型3株均為腫瘤醫(yī)院住院患者分離株；一附院與腫瘤醫(yī)院患者STS基因分型已知型占全部例數(shù)的比例分別為87.5%、53.7%,經(jīng)卡方檢驗(yàn)差異有統(tǒng)計(jì)學(xué)意義。RAPD結(jié)果顯示21例分離株的遺傳相似系數(shù)Sij在0.615-0.923之間；從聚類(lèi)分析樹(shù)可將21例B.h可劃分為8個(gè)枝系。(3)MTT法檢測(cè)藥敏實(shí)驗(yàn)結(jié)果顯示甲硝唑、替硝唑、阿奇霉素、諾氟沙星4種藥物均對(duì)B.h有抑制作用,且隨著濃度的增加(10、20、40、80μg/ml)其平均細(xì)胞抑制率是在不斷增加；經(jīng)R語(yǔ)言軟件計(jì)算4種藥物在24h、48h、72h時(shí)間段的IC50值,加入藥物24h后,甲硝唑、替硝唑、阿奇霉素、諾氟沙星的IC50值分別為159.77、80.47、162.5、77.27μg/ml;加入藥物48h后,IC50值分別為1147、440.9、2847、169.2μg/ml;加入藥物72h后,IC50值分別為2146、1967、8031、7895μg/ml。結(jié)論(1)住院患者感染率較正常人群感染率高,腫瘤患者感染率高于非腫瘤患者和正常人群感染率,胃腸科室住院患者感染率高于非胃腸科室住院患者感染率。(2)在住院患者中存在1、2、3、4、6、7型,未見(jiàn)5型,且2型為優(yōu)勢(shì)基因型。(3)21例未知基因型基因結(jié)構(gòu)復(fù)雜,遺傳差異明顯。(4)諾氟沙星對(duì)抑制B.h的生長(zhǎng)與繁殖作用明顯。
[Abstract]:Objective to understand the infection of Blastocystis hominis (Blastocystis hominis, B.h) in two hospital inpatients and determine the genotype of B.h infected strains in hospitalized patients, to reveal the genetic distance and genetic polymorphism of unknown genotype B.h isolates, and to explore the effect of metronidazole, nitazazole, azithromycin and norfloxacin on B.h. Methods the fresh feces of hospitalized patients were collected from the First Affiliated Hospital of Guangxi (the First Affiliated Hospital of Guangxi Medical University, the tumor hospital affiliated to Guangxi Medical University, the following abbreviated hospital and cancer hospital). The modified acid ether precipitation method, the microscopic examination diagnosis, and the R language soft parts were used to analyze the hospitalized patients and the normal population and the patients with gastrointestinal diseases. In patients with non gastrointestinal diseases, the B.h infection rates of B.h were collected between the sexes, professions, ages, ethnic groups and education levels. The positive insect body was collected and the positive insect body DNA was extracted at 37 C, and the genomic DNA was used as a template, and 7 STS specific primers were used to identify the genotyping by PCR amplification; the unknown genotypes were divided. The isolated strain was used as the research object to construct a strip map with RAPD marker technique. The genetic diversity of the isolates was analyzed by NTSYSpc software and the cluster analysis was constructed. Finally, the drug sensitivity of B.h to metronidazole, tinidazole, azithromycin, norfloxacin was detected by MTT, and the effect of 4 drugs on B.h was observed. Results (1) two medical treatments. The total positive rate of the hospitalized patients was 2800 cases, the positive rate of the total positive rate was 22.59% in the 36.6%. hospital and the tumor hospital. Compared with the normal population, the infection rate of the hospitalized patients was higher than that of the normal population (P=0.000). The rate of infection in the tumor patients and the non tumor patients was compared with the normal population (P=0.000), and the difference was statistically significant (P=0.000). The difference of infection rate between the two groups was statistically significant (P=0.000). According to the educational level, occupation division, two hospital junior high school and below education patients (P=0.000), the infection rate of the farmers was statistically significant (P=0.000) compared with the other cultural degree patients (P=0.000); according to sex, age, national group, two hospitals There was no significant difference in infection rate between each group (P0.05). (2) the results of STS genotyping showed that there were 57 specimens and 36 specimens were classified as simple 1, 20, only 3, 3, only 4, 3, purely 6 5, pure 7. The isolated strains of hospitalized patients and 3 strains of type 3 were all isolated strains of inpatients in the cancer hospital. The proportion of the known STS genotypes of the patients in the hospital and the cancer hospital was 87.5% and 53.7% respectively. The difference of the chi square test was statistically significant.RAPD results showed that the genetic similarity coefficient Sij of the 21 isolates was between 0.615-0.923; The cluster analysis tree could divide 21 B.h cases into 8 branches. (3) the test results of MTT assay showed that metronidazole, tinidazole, azithromycin, and norfloxacin had inhibitory effects on B.h, and the average cell inhibition rate was increasing with the increase of concentration (10,20,40,80 mu g/ml); 4 drugs were calculated by R language software in 24h, 4 After adding drug 24h, the IC50 value of metronidazole, tinidazole, azithromycin and norfloxacin was 159.77,80.47162.5,77.27 g/ml respectively after adding the IC50 value of the drug 24h, and the IC50 value was 1147440.92847169.2 u g/ml, respectively after adding drug 48h, after adding drug 72h, the value was 2146196780317895 micron conclusion (1) in hospitalized patients. The infection rate of the normal population is high, the infection rate of the tumor patients is higher than that of the non tumor patients and the normal population. The infection rate of the hospitalized patients in the gastrointestinal department is higher than that in the non gastrointestinal department. (2) there is a 1,2,3,4,6,7 type in the hospitalized patients, no type 5, and the 2 type is the dominant genotype. (3) 21 cases of unknown genotypes are complex and genetic difference. (4) norfloxacin significantly inhibited the growth and reproduction of B.h.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R446.5

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本文編號(hào)：1937505