【作者】 韩晶；

【导师】 宫玉香；

【作者基本信息】 青岛大学 ， 病原生物学， 2008， 硕士

【摘要】 目的:采用瑞氏-吉姆萨染色法、巢式PCR法检测耶氏肺孢子虫,比较这两种方法在耶氏肺孢子虫肺炎诊断中的敏感性和稳定性,并结合病人的临床表现探讨肺孢子虫肺炎的临床诊断方法。方法:收集艾滋病、肾移植、肺癌、支气管炎等患者的痰液、气管分泌物、支气管肺泡灌洗液(BLA)或肺组织标本,采用瑞氏-吉姆萨染色法检测痰、BALF涂片和肺印片等标本中的耶氏肺孢子虫;以Pc-ITS为引物,对肺组织、BALF和痰标本,采用巢式PCR法检测耶氏肺孢子虫DNA。通过文献检索统计国内PCP发病情况:性别及年龄比例、发病特点、地区分布等,结合本研究发现的PCP病人资料,分析总结耶氏肺孢子虫肺炎的临床诊断方法。结果:瑞氏-吉姆萨染色法对该虫的检出率为32.39%,巢氏PCR法对该虫DNA的检出率为61.97%,二者检出率比较,x~2=11.30,p＜0.01,有显著意义。文献检索发现2001～2007年国内PCP病例,以AIDS并发PCP为主(82.10%);男性多于女性;死亡率13.08%,死亡病人中AIDS并发PCP病人占97.37%;PCP发病地区主要在河南、广西、湖南、广东等地;本研究确诊耶氏肺孢子虫肺炎6例,男5例,女1例,治疗后均好转出院。结论:瑞氏-吉姆萨染色法,操作简便,但敏感性偏低。巢式PCR法检测耶氏肺孢子虫敏感性高于瑞氏-吉姆萨染色法,其特异性和敏感性也优于普通的单一引物PCR法,本研究首次在青岛地区采用巢氏PCR法检出AIDS病人并发耶氏肺孢子虫,所用引物PC-ITS对耶氏肺孢子虫高度敏感,适用于临床病人的检测,结果准确快捷,值得推广。我国目前PCP发病人群以AIDS并发PCP为主,根据国内外资料综述及我们在本研究中对PCP阳性病人的分析,符合以下标准可诊断为AIDS并发PCP:①HIV感染;②CD4+小于200/μl;③有发热、咳嗽,咳痰,伴呼吸急促、进行性呼吸困难、紫绀,血氧饱和度下降;④胸片示间质性肺炎改变,或/及CT肺部示毛玻璃状改变;⑤复方新诺明治疗有效。更多还原

【Abstract】 Objective: To compare the sensitivity and stability of Wright-Giemsa’s stained, nested PCR asssy(PC-ITS-PCR) to detect the Pneumocystis Jiroveci in patients with Pneumocystis Jiroveci Pneumonia, and to explore the diagnosis of PCP concerning clinical manifestation.Methods: Collect the sputum, tracheal discharge, bronchoalveolar lavage fluid and specimen of lung tissue from patients with AIDS, kidney transplant, lung cancer and bronchitis, Wright-Giemsa’s stained was used to detect the organism in sputum, lung impression preparation and bronchoalveolar lavage fluid(BALF). Pneumocystis jiroveci DNA in sputum, lung tissue and BALF were detected by nested PCR method using PC-ITS primers. Retrieve incidence of PCP in china through literature involving portion of gender and age, feature and endemic distribution incorporated the data of this research to explore clinical diagnosis of PCP.Result: In this study, detection rate of Wright-Giemsa’s stained is 32.39%, while the sensitivity of PCR is 61.97%. (x~2=11.30, p<0.01),there is significant difference.. Through literature retrieval we find domestic PCP cases between 2001-2007 are mostly AIDS complicating PCP (82.10%), male are more than female and mortality is 13.08%. Patients with AIDS complicating PCP account for 97.37% of death. Invasion regions are mostly Henan, Guangxi, Hunan and Guangdong; 6 cases of Pneumocystis Jiroveci Pneumonia are confirmed in this research (male 5,femal l)and all are improved and discharged after treatment.Conclusion: The Wright-Giemsa’s stained to detect PC is simple t but the specificity is low while nested PC-ITS-PCR method has higher sensitivity than that of the Wright-Giemsa’s stained and has higher sensitivity and specificity than PCR with single primer. Nested PC-ITS-PCR is the first time used to detect PCP in the research in Patients with AIDS in Qingdao while premier PC-ITS is highly sensitive to PC and applicable to detect clinical patients. The result is accurate and fast, so nested PC-ITS-PCR should be recommended. In china, most cases of PCP are Patients with AIDS complicating. According to data at present and our analysis in this research, patients with he following findings can be diagnosed AIDS complicating PCP (a) HIV infection, (b) CD4~+<200/μl (c)fever, cough, expectoration accompanying tachypnea, progressive dyspnea, cyanosis, decreasing saturation of blood oxygen (d) X-ray of chest revealing interstitial pneumonia or/and CT showing ground glass change of lung. (e) effective treatment by trimethoprirn-sulfamethoxazole更多还原