本文選題：肺孢子肺炎 + HIV陰性免疫力低下者��； 參考：《重慶醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：這項(xiàng)研究的目的是通過檢測重慶地區(qū)人類免疫缺陷病毒(Human immunodeficiency virus,HIV)陰性、免疫力低下患者痰液中的肺孢子（Pneumocystis carinii, PC）來探討該類患者PC的感染率；通過比較不同檢測方法對PC的檢測效率，從而探討實(shí)時(shí)熒光定量PCR（Real-time fluorescence quantitative PCR, r-PCR）在臨床上對HIV陰性免疫力低下患者合并肺孢子肺炎(Pneumocystis carinii pneumonia, PCP)的診斷價(jià)值；通過對PC內(nèi)轉(zhuǎn)錄間隔區(qū)(Internal transcribed spacer, ITS)基因進(jìn)行序列分析來對重慶地區(qū)PC的基因多態(tài)性進(jìn)行初步分析。 方法：收集重慶地區(qū)71例HIV陰性免疫力低下患者的痰液標(biāo)本，其中結(jié)締組織疾病患者23例，腫瘤放化療患者21例，血液系統(tǒng)疾病患者15例，肝移植術(shù)后患者7例，腎移植術(shù)后患者5例。以ITS基因?yàn)槟康幕颍謩e運(yùn)用六亞甲基四胺銀（Gomort methenamine silver stain,GMS)染色法、巢式PCR法和r-PCR法檢測標(biāo)本中的PC，將兩種PCR法檢測均為陽性的標(biāo)本的PCR產(chǎn)物送予基因序列測定，然后進(jìn)行PCITS基因多態(tài)性分析。 結(jié)果：（1）71名患者中，痰液標(biāo)本經(jīng)GMS染色后，僅有2例結(jié)果為陽性，，經(jīng)巢式PCR檢測后，25例結(jié)果為陽性，巢式PCR的檢出率顯著高于GMS染色法；PC感染率為35.21%，其中腫瘤放化療后患者、血液系統(tǒng)疾病患者、腎移植患者、肝移植患者、結(jié)締組織疾病患者感染率分別為33.33%、26.67%、40.00%、42.86%、39.13%，各病例組間感染率差異無顯著性，但本組病例PC感染率顯著高于同期國外報(bào)道，感染率同2009年國內(nèi)報(bào)道值接近。（2）根據(jù)臨床PCP診斷結(jié)果，巢式PCR假陽性率較高為36.00%，其特異度為83.01%，靈敏度為88.89%，陽性預(yù)測值為64.00%，當(dāng)用103copes/ml為判斷r-PCR結(jié)果陰陽性的閾值時(shí)，r-PCR的靈敏度（83.33%）同巢式PCR無顯著差異性，但它的特異度（98.11%）和陽性預(yù)測值（93.75%）均顯著高于巢式PCR；臨床診斷PCP患者中r-PCR檢測出PC的基因載量顯著高于非臨床診斷PCP患者。（3）通過對15例陽性標(biāo)本的PCR反應(yīng)物進(jìn)行測序，獲得15條ITS序列，其中ITS1共獲得了4個(gè)基因型（B,M,E,K）和11條新序列，獲得了ITS2的3個(gè)基因型（g,i,b）和12條新序列，5.8SrRNA得到了7條新序列，經(jīng)過組合，得到了1個(gè)已知的ITS基因型（Bi）和14個(gè)新組合序列，而所有新序列與國外報(bào)道的原型比較，存在6~21個(gè)堿基差異，差異主要表現(xiàn)為堿基缺失和堿基替換。 結(jié)論：重慶地區(qū)HIV陰性免疫力低下患者群PC感染率較高，臨床醫(yī)師應(yīng)增強(qiáng)對該類患者是否合并PC感染的警覺性；在HIV陰性免疫力低下患者中，使用痰液為標(biāo)本時(shí)，r-PCR比巢式PCR更有助于準(zhǔn)確而快速地診斷PCP，并且可以區(qū)分PC定植與感染從而指導(dǎo)臨床診治；重慶區(qū)域中，PC-ITS基因具有基因多態(tài)性，且可能有新的基因分型。
[Abstract]:Objective: the aim of this study was to investigate the infection rate of human immunodeficiency virus (immunodeficiency virus) in the sputum of patients with low immunity by detecting the Pneumocystis carinii (PCS) in the sputum of patients with human immunodeficiency virus (immunodeficiency) in Chongqing area. By comparing the efficiency of different detection methods for PC, the diagnostic value of real-time fluorescent quantitative PCR(Real-time fluorescence quantitative PCR, r-PCR for Pneumocystis carinii pneumonia, PCP) in HIV negative immunocompromised patients with Pneumocystis was studied. The polymorphism of internal transcribed spacer, ITS) gene of PC in Chongqing was analyzed by sequence analysis of internal transcriptional spacer region of PC. Methods: sputum samples were collected from 71 patients with HIV negative immunosuppression in Chongqing, including 23 patients with connective tissue disease, 21 patients with tumor radiotherapy and chemotherapy, 15 patients with hematological diseases and 7 patients with liver transplantation. 5 cases after renal transplantation. The ITS gene was detected by Hexamethylenetetramine Gomort methenamine silver stencils staining method, nested PCR method and r-PCR method respectively. The PCR products of both positive samples detected by PCR method were sent to the gene sequence analysis. Then the polymorphism of PCITS gene was analyzed. Results only 2 sputum samples were positive after GMS staining, and 25 cases were positive after nested PCR examination. The positive rate of nested PCR was significantly higher than that of GMS staining (35.21%). The infection rates of patients with hematological diseases, renal transplantation, liver transplantation and connective tissue diseases were 33.33 and 26.676.67 respectively. There was no significant difference in infection rate among different groups, but the PC infection rate in this group was significantly higher than that reported abroad in the same period. The infection rate was close to the reported value in 2009. (2) according to the clinical PCP diagnosis results, The false positive rate of nested PCR was 36.00, the specificity was 83.01, the sensitivity was 88.89, and the positive predictive value was 64.00. The sensitivity of r-PCR was 83.33 when 103copes/ml was used as the threshold to judge the negative and positive results of r-PCR. But its specificity (98.11) and positive predictive value (93.755) were significantly higher than those of nested PCP. The gene load of PC detected by r-PCR in clinically diagnosed PCP patients was significantly higher than that in non-clinically diagnosed PCP patients. 15 ITS sequences were obtained, of which 4 genotypes were obtained from ITS1, 11 new sequences, 3 genotypes of ITS2 and 5. 8s rRNA, and 7 new sequences were obtained by combination. A known ITS genotype (Bibi) and 14 new combination sequences were obtained. Compared with the reported prototypes, there were 6 ~ 21 nucleotide differences in all the new sequences. The differences were mainly expressed as base deletion and base substitution. Conclusion: the infection rate of PC in HIV negative immunocompromised patients in Chongqing area is relatively high. Clinicians should enhance their awareness of whether these patients are complicated with PC infection, and in patients with HIV negative immunosuppression, Using sputum as a sample, r-PCR is more helpful than nested PCR in the accurate and rapid diagnosis of PCR, and it can distinguish between PC colonization and infection to guide clinical diagnosis and treatment. In Chongqing region, PC-ITS gene has gene polymorphism and may have new genotyping.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R512.91

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

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2 安亦軍,黃敏君,郭增柱;PCR及GMS染色法對肺孢子蟲肺炎的臨床診斷價(jià)值[J];中國寄生蟲病防治雜志;2005年04期

本文編號(hào)：1920727