广西两所三甲医院住院患者人芽囊原虫感染现状、基因分型及体外药敏试验

发布时间：2018-05-26 14:07

本文选题：人芽囊原虫 + 感染情况　；参考：《广西医科大学》2015年硕士论文

【摘要】：目的了解广西两所三甲医院住院患者人芽囊原虫(Blastocystis hominis,B.h)的感染情况；确定住院患者B.h感染株的基因分型,揭示未知基因型B.h分离株的遗传距离及遗传多态性；探讨甲硝唑、替硝唑、阿奇霉素、诺氟沙星4种药物对B.h的作用效果。方法收取广西三甲医院(广西医科大学第一附属医院、广西医科大学附属肿瘤医院,以下分别简称一附院和肿瘤医院)住院患者新鲜粪便,采用改良酸醚沉淀法,镜检定性诊断,用R语言软件分析住院患者与正常人群、胃肠疾病患者与非胃肠疾病患者、两所医院住院患者性别、职业、年龄、民族及受教育水平各组间B.h感染率。收集经鉴定的阳性虫体,37℃厌氧培养,提取阳性虫体DNA；以基因组DNA为模板,采用7对STS特异性引物进行PCR扩增明确其基因分型；把未知基因型分离株作为研究对象应用RAPD标记技术构建条带图谱,用NTSYSpc软件对分离株的遗传多样性进行聚类分析,构建聚类分析图。最后采用MTT法检测B.h对甲硝唑、替硝唑、阿奇霉素、诺氟沙星的药敏反应,观察4种药物对B.h的作用效果。结果(1)两所医院共检查住院患者2800例,总阳性率为36.6%。附院与肿瘤医院阳性率分别为22.59%、43.24%。与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)；肿瘤患者与非肿瘤患者、正常人群感染率相比,差异有统计学意义(P=0.000)；按照胃肠科室与非胃肠科室分组,两组感染率差异有统计学意义(P=0.000)；按照文化程度、职业划分,两所医院初中及以下学历患者(P=0.000)、农民患者感染率较其他文化程度患者感染率差异有统计学意义(P=0.000)；按照性别、年龄、民族分组,两所医院各组间感染率差异没有统计学意义(P0.05)。(2)STS基因分型结果显示,共57例标本,有36例标本被分型,其中单纯1型2株、单纯2型20株、单纯3型3株、单纯4型3株、单纯6型5株、单纯7型3株,剩余21例为未知基因型；1型、4型、7型共扩增出8株,且8株均为一附院住院患者分离株,3型3株均为肿瘤医院住院患者分离株；一附院与肿瘤医院患者STS基因分型已知型占全部例数的比例分别为87.5%、53.7%,经卡方检验差异有统计学意义。RAPD结果显示21例分离株的遗传相似系数Sij在0.615-0.923之间；从聚类分析树可将21例B.h可划分为8个枝系。(3)MTT法检测药敏实验结果显示甲硝唑、替硝唑、阿奇霉素、诺氟沙星4种药物均对B.h有抑制作用,且随着浓度的增加(10、20、40、80μg/ml)其平均细胞抑制率是在不断增加；经R语言软件计算4种药物在24h、48h、72h时间段的IC50值,加入药物24h后,甲硝唑、替硝唑、阿奇霉素、诺氟沙星的IC50值分别为159.77、80.47、162.5、77.27μg/ml;加入药物48h后,IC50值分别为1147、440.9、2847、169.2μg/ml;加入药物72h后,IC50值分别为2146、1967、8031、7895μg/ml。结论(1)住院患者感染率较正常人群感染率高,肿瘤患者感染率高于非肿瘤患者和正常人群感染率,胃肠科室住院患者感染率高于非胃肠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,未见5型,且2型为优势基因型。(3)21例未知基因型基因结构复杂,遗传差异明显。(4)诺氟沙星对抑制B.h的生长与繁殖作用明显。
[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.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.5

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