一.干擾素釋放試驗(yàn)對(duì)于結(jié)核性腦膜炎的診斷價(jià)值以及診斷模型的建立 二.結(jié)核性腦膜炎差異miRNA的初步篩選
發(fā)布時(shí)間：2018-09-18 17:42
【摘要】：TB目前仍然是嚴(yán)重威脅人類生命的最主要感染性疾病，中國(guó)2010年全國(guó)結(jié)核病流行病學(xué)調(diào)查顯示肺結(jié)核病例約500萬，每年新發(fā)病例達(dá)100萬。結(jié)核性腦膜炎（tuberculous meningitis，TBM）的發(fā)病率雖然僅占結(jié)核病的1%-2%，但是其死亡率和致殘率卻高于40%，，高死亡率主要是缺乏敏感特異的診斷手段所致。因此，尋找一種新的敏感特異的診斷方法，提高TBM的診斷水平，是降低TBM死亡率和致殘率的關(guān)鍵。 γ-干擾素釋放試驗(yàn)（interferon gamma releasing assay，IGRAs）根據(jù)特異性抗原刺激后機(jī)體效應(yīng)T淋巴細(xì)胞產(chǎn)生的γ-干擾素來判斷診斷結(jié)核感染的免疫學(xué)方法，由于不能區(qū)分結(jié)核潛伏感染（latent tuberculosis infection，LTBI）和活動(dòng)性結(jié)核病（active tuberculosis，ATB），而致其在TB高負(fù)擔(dān)國(guó)家診斷價(jià)值受限。結(jié)核分枝桿菌（Mycobacterium Tuberculosis，M.TB）感染部位具有隔室化的特點(diǎn)，即抗原特異性T淋巴細(xì)胞被募集到局部感染部位，感染部位如腦脊液（cerebrospinal fluid，CSF）的T-SPOT.TB有可能獲得比外周血（peripheral blood，PB) T-SPOT.TB更高的診斷準(zhǔn)確性。目前我國(guó)尚沒有CSF T-SPOT.TB檢測(cè)診斷TBM的相關(guān)數(shù)據(jù)。 Thwaites GE等2002年通過五種臨床特征建立了預(yù)測(cè)TBM的越南模型。模型包含五項(xiàng)內(nèi)容：年齡、病程、血常規(guī)白細(xì)胞計(jì)數(shù)、CSF白細(xì)胞計(jì)數(shù)和CSF中性粒細(xì)胞比例。越南模型主要是鑒別TBM和細(xì)菌性腦膜炎，而在我國(guó)細(xì)菌性腦膜炎發(fā)病率相對(duì)較低，因此這一模型并不一定符合我國(guó)腦膜炎的發(fā)病現(xiàn)狀，還需要建立適合我國(guó)的一種新的診斷模型。由于PB T-SPOT.TB和CSFT-SPOT.TB可能具有較高的診斷準(zhǔn)確性，將這兩項(xiàng)指標(biāo)以及其他臨床檢測(cè)指標(biāo)聯(lián)合，建立診斷模型，有可能獲得較好的診斷準(zhǔn)確性。 本課題最終納入TBM31例和non-TBM40例進(jìn)行PB T-SPOT.TB和CSFT-SPOT.TB，兩種檢測(cè)的敏感性分別為93.6%（29/31,95%CI：79.3%-98.2%）和53.3%（16/30，95%CI：36.1%-70.0%），特異性分別為75.0%（30/40，95%CI：59.8%-85.8%）和94.8%（37/39，95%CI：83.1%-98.6%），診斷準(zhǔn)確性分別為83.7%（59/71,72.7%-90.1%）和76.8%（53/69，95%CI：65.6%-85.2%）。PB T-SPOT診斷的敏感性高于CSF T-SPOT.TB，而CSF T-SPOT.TB的特異性高于PBT-SPOT.TB。將這兩項(xiàng)指標(biāo)和TBM、non-TBM之間存在差異的CSF蛋白、CSF腺苷脫氨酶和發(fā)病時(shí)間等共五項(xiàng)指標(biāo)輸入支持向量機(jī)（SVM），優(yōu)選出的最具診斷意義的是PB T-SPOT.TB和CSF T-SPOT.TB，用這兩項(xiàng)指標(biāo)確立決策樹、建立聯(lián)合診斷模型，得到聯(lián)合診斷模型的敏感性和特異性分別為90.0%（27/30，95%CI：27/30）和94.9%（37/39，95%CI：83.1%-98.6%），其診斷準(zhǔn)確性為92.8%（64/69，95%CI：84.8%-96.9%），高于PB T-SPOT.TB的和CSF T-SPOT.TB。而從本課題納入的病例分析，越南模型驗(yàn)證的特異性僅為5%。 本研究表明PB T-SPOT.TB由于敏感性較高，有可能作為排除TBM診斷的一種方法，CSF T-SPOT.TB由于特異性較高，有可能作為目前納入TBM診斷的一種方法。越南模型并不適用于我國(guó)TBM的診斷和鑒別診斷，而根據(jù)PBT-SPOT.TB和CSF T-SPOT.TB建立的聯(lián)合診斷模型對(duì)于結(jié)核性腦膜炎的診斷具有較高的診斷價(jià)值，但是無論單一的PB T-SPOT.TB和CSF T-SPOT.TB檢測(cè)，或及聯(lián)合診斷模型的意義，都需要進(jìn)一步進(jìn)行大樣本前瞻性實(shí)驗(yàn)加以驗(yàn)證。 miRNA是近年來生命科學(xué)研究的熱點(diǎn)，某些miRNA或可作為腫瘤、自身免疫性疾病診斷的生物標(biāo)識(shí)。然而，miRNA作為生物標(biāo)識(shí)在細(xì)菌感染性疾病尤其是TB中的研究相對(duì)較少。由于這種小分子在細(xì)胞的生長(zhǎng)、分化、凋亡、代謝以及病毒感染、腫瘤的發(fā)生等生理病理過程中具有重要作用，因此推測(cè)其在TBM發(fā)病的免疫學(xué)機(jī)制、疾病的進(jìn)展等方面也可能具有一定的意義，有可能做為TBM診斷的生物標(biāo)識(shí)。診斷手段的缺乏是導(dǎo)致TBM的死亡率多年來高居不下的首要原因，如果找到特異表達(dá)的miRNA，則可以提高TBM的診斷水平。 本研究入選TBM組8例，non-TBM（病毒性腦膜炎）組8例和健康對(duì)照6例，應(yīng)用Agilent芯片篩選得到11種差異miRNA，具體為hsa-miR-126-3p/5p，hsa-miR-130a-3p， hsa-miR-151a-3p/5p， hsa-miR-199a-3p， hsa-miR-221-3p，hsa-miR-4291，hsa-miR-584-5p，hsa-miR-6127，hsa-miR-744-5p。通過靶基因預(yù)測(cè)和GO分析發(fā)現(xiàn)上述差異miRNA的靶基因的功能主要是參與轉(zhuǎn)錄過程、DNA與RNA的代謝過程等；Pathway通路分析顯示miRNA的靶基因主要是參與腫瘤通路、MAPK信號(hào)通路、Wnt通路等。今后的工作中將進(jìn)一步篩選病例擴(kuò)大樣本量進(jìn)行RT-PCR驗(yàn)證，以確定差異miRNA是否具有診斷TBM的意義，同時(shí)要進(jìn)一步的研究以證實(shí)這些差異miRNA是否與炎癥反應(yīng)以及TB、TBM發(fā)病的免疫機(jī)制相關(guān)。
[Abstract]:Tuberculous meningitis (TBM) is still the most serious infectious disease that threatens human life. The epidemiological survey of tuberculosis in China in 2010 showed that about 5 million cases of tuberculosis and 1 million new cases are reported every year. Although the incidence of tuberculous meningitis (TBM) is only 1% - 2% of tuberculosis, its mortality and disability rate are high. At 40%, the high mortality rate is mainly due to the lack of sensitive and specific diagnostic methods. Therefore, finding a new sensitive and specific diagnostic method to improve the diagnostic level of TBM is the key to reduce the mortality and disability rate of TBM.
Interferon gamma release assay (IGRAs) is an immunological method for the diagnosis of tuberculosis infection based on interferon-gamma produced by effector T lymphocytes stimulated by specific antigens. It can not distinguish latent tuberculosis infection (LTBI) from active tuberculosis (active tuberculosis). Mycobacterium tuberculosis (M. TB) infection site has the characteristics of compartmentalization, that is, antigen-specific T lymphocytes are recruited to locally infected sites, infection sites such as cerebrospinal fluid (CSF) T-SPOT. TB may obtain more than peripheral blood (periph). The diagnostic accuracy of PB T-SPOT.TB is higher than that of PB T-SPOT.TB.
Thwaites GE et al. established a Vietnamese model for predicting TBM through five clinical features in 2002. The model consists of five items: age, course of disease, blood routine white blood cell count, CSF white blood cell count and CSF neutrophil ratio. Because PB T-SPOT.TB and CSFT-SPOT.TB may have higher diagnostic accuracy, it is possible to combine these two indicators with other clinical detection indicators to establish a diagnostic model, which may obtain better diagnostic accuracy. Sex.
The sensitivity of PB T-SPOT.TB and CSFT-SPOT.TB were 93.6% (29/31, 95% CI: 79.3% - 98.2%) and 53.3% (16/30, 95% CI: 36.1% - 70.0%) respectively, and the specificity was 75.0% (30/40, 95% CI: 59.8% - 85.8%) and 94.8% (37/39, 95% CI: 83.1% - 98.6%) respectively. The diagnostic accuracy was 57.7% (59/39, 95% CI: 83.1% - 98.6%). The sensitivity of PBT-SPOT was higher than that of CSF T-SPOT.TB, and the specificity of CSF T-SPOT.TB was higher than that of PBT-SPOT. PB T-SPOT.TB and CSF T-SPOT.TB were used to establish decision tree and joint diagnosis model. The sensitivity and specificity of the combined diagnosis model were 90.0% (27/30, 95% CI: 27/30) and 94.9% (37/39, 95% CI: 83.1% - 98.6%) respectively. The diagnostic accuracy was 92.8% (64/69, 95% CI: 84.8% - 96.9%) higher than that of PB T-SPOT. The specificity of B model and CSF T-SPOT.TB. was only 5%. for the case analysis.
This study indicates that PBT-SPOT.TB may be used as a method to exclude TBM because of its high sensitivity and specificity. CSF T-SPOT.TB may be used as a method to diagnose TBM at present. Vietnam model is not suitable for the diagnosis and differential diagnosis of TBM in China, but the combined diagnosis based on PBT-SPOT.TB and CSF T-SPOT.TB. The severed model has a high diagnostic value for tuberculous meningitis, but the significance of single PB T-SPOT.TB and CSF T-SPOT.TB test, or combined diagnosis model, needs to be further validated by large-sample prospective experiments.
MicroRNAs are the hotspot of life science research in recent years. Some microRNAs may be used as biomarkers for the diagnosis of tumors and autoimmune diseases. However, there are relatively few studies on microRNAs as biomarkers in bacterial infectious diseases, especially in TB. It may be a biomarker for the diagnosis of TBM. The lack of diagnostic tools is the primary cause of the high mortality rate of TBM over the years. If the specific expression of microRNAs can be found, it can be used as a biomarker for the diagnosis of TBM. NA can improve the diagnostic level of TBM.
In this study, 8 TBM patients, 8 non-TBM (viral meningitis) patients, 8 non-TBM (viral meningitis) patients and 6 healthy controls were enrolled, 11 different microRNAs were screened by Agilent chip, including hsa-microRNA-126-3p/5p, hsa-microRNA-130a-3p, hsa-microRNA-151a-3p/5p, hsa-microRNA-199a-199a-3p, hsa-microRNA-199a-199a-199a-3p, hsa-microRNA-4291, hsa-microarray-microRNA-584-584-5p, hsa-5p, hsa-microRNA-614-5p, hsa-61a-61a-61a-61a-61a P. Pre-gene by Target The results of GO analysis showed that the target genes of these differentially expressed microRNAs were mainly involved in transcription, DNA and RNA metabolism, and Pathway pathway analysis showed that the target genes of microRNAs were mainly involved in tumor pathway, MAPK signaling pathway and Wnt pathway. To determine whether differentially expressed microRNAs have diagnostic significance for TBM, further studies are needed to confirm whether these differentially expressed microRNAs are associated with inflammation and the immune mechanism of TB and TBM.
【學(xué)位授予單位】：北京市結(jié)核病胸部腫瘤研究所
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R529.3
本文編號(hào)：2248646
[Abstract]:Tuberculous meningitis (TBM) is still the most serious infectious disease that threatens human life. The epidemiological survey of tuberculosis in China in 2010 showed that about 5 million cases of tuberculosis and 1 million new cases are reported every year. Although the incidence of tuberculous meningitis (TBM) is only 1% - 2% of tuberculosis, its mortality and disability rate are high. At 40%, the high mortality rate is mainly due to the lack of sensitive and specific diagnostic methods. Therefore, finding a new sensitive and specific diagnostic method to improve the diagnostic level of TBM is the key to reduce the mortality and disability rate of TBM.
Interferon gamma release assay (IGRAs) is an immunological method for the diagnosis of tuberculosis infection based on interferon-gamma produced by effector T lymphocytes stimulated by specific antigens. It can not distinguish latent tuberculosis infection (LTBI) from active tuberculosis (active tuberculosis). Mycobacterium tuberculosis (M. TB) infection site has the characteristics of compartmentalization, that is, antigen-specific T lymphocytes are recruited to locally infected sites, infection sites such as cerebrospinal fluid (CSF) T-SPOT. TB may obtain more than peripheral blood (periph). The diagnostic accuracy of PB T-SPOT.TB is higher than that of PB T-SPOT.TB.
Thwaites GE et al. established a Vietnamese model for predicting TBM through five clinical features in 2002. The model consists of five items: age, course of disease, blood routine white blood cell count, CSF white blood cell count and CSF neutrophil ratio. Because PB T-SPOT.TB and CSFT-SPOT.TB may have higher diagnostic accuracy, it is possible to combine these two indicators with other clinical detection indicators to establish a diagnostic model, which may obtain better diagnostic accuracy. Sex.
The sensitivity of PB T-SPOT.TB and CSFT-SPOT.TB were 93.6% (29/31, 95% CI: 79.3% - 98.2%) and 53.3% (16/30, 95% CI: 36.1% - 70.0%) respectively, and the specificity was 75.0% (30/40, 95% CI: 59.8% - 85.8%) and 94.8% (37/39, 95% CI: 83.1% - 98.6%) respectively. The diagnostic accuracy was 57.7% (59/39, 95% CI: 83.1% - 98.6%). The sensitivity of PBT-SPOT was higher than that of CSF T-SPOT.TB, and the specificity of CSF T-SPOT.TB was higher than that of PBT-SPOT. PB T-SPOT.TB and CSF T-SPOT.TB were used to establish decision tree and joint diagnosis model. The sensitivity and specificity of the combined diagnosis model were 90.0% (27/30, 95% CI: 27/30) and 94.9% (37/39, 95% CI: 83.1% - 98.6%) respectively. The diagnostic accuracy was 92.8% (64/69, 95% CI: 84.8% - 96.9%) higher than that of PB T-SPOT. The specificity of B model and CSF T-SPOT.TB. was only 5%. for the case analysis.
This study indicates that PBT-SPOT.TB may be used as a method to exclude TBM because of its high sensitivity and specificity. CSF T-SPOT.TB may be used as a method to diagnose TBM at present. Vietnam model is not suitable for the diagnosis and differential diagnosis of TBM in China, but the combined diagnosis based on PBT-SPOT.TB and CSF T-SPOT.TB. The severed model has a high diagnostic value for tuberculous meningitis, but the significance of single PB T-SPOT.TB and CSF T-SPOT.TB test, or combined diagnosis model, needs to be further validated by large-sample prospective experiments.
MicroRNAs are the hotspot of life science research in recent years. Some microRNAs may be used as biomarkers for the diagnosis of tumors and autoimmune diseases. However, there are relatively few studies on microRNAs as biomarkers in bacterial infectious diseases, especially in TB. It may be a biomarker for the diagnosis of TBM. The lack of diagnostic tools is the primary cause of the high mortality rate of TBM over the years. If the specific expression of microRNAs can be found, it can be used as a biomarker for the diagnosis of TBM. NA can improve the diagnostic level of TBM.
In this study, 8 TBM patients, 8 non-TBM (viral meningitis) patients, 8 non-TBM (viral meningitis) patients and 6 healthy controls were enrolled, 11 different microRNAs were screened by Agilent chip, including hsa-microRNA-126-3p/5p, hsa-microRNA-130a-3p, hsa-microRNA-151a-3p/5p, hsa-microRNA-199a-199a-3p, hsa-microRNA-199a-199a-199a-3p, hsa-microRNA-4291, hsa-microarray-microRNA-584-584-5p, hsa-5p, hsa-microRNA-614-5p, hsa-61a-61a-61a-61a-61a P. Pre-gene by Target The results of GO analysis showed that the target genes of these differentially expressed microRNAs were mainly involved in transcription, DNA and RNA metabolism, and Pathway pathway analysis showed that the target genes of microRNAs were mainly involved in tumor pathway, MAPK signaling pathway and Wnt pathway. To determine whether differentially expressed microRNAs have diagnostic significance for TBM, further studies are needed to confirm whether these differentially expressed microRNAs are associated with inflammation and the immune mechanism of TB and TBM.
【學(xué)位授予單位】：北京市結(jié)核病胸部腫瘤研究所
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R529.3
【參考文獻(xiàn)】
相關(guān)期刊論文 前4條
1 徐文鴻;陳益定;胡躍;余捷凱;王連聰;鄭樹;張?zhí)K展;;基于支持向量機(jī)的大腸黏液腺癌術(shù)前血清蛋白質(zhì)標(biāo)志物檢測(cè)及分析[J];細(xì)胞生物學(xué)雜志;2008年06期
2 全國(guó)結(jié)核病流行病學(xué)抽樣調(diào)查技術(shù)指導(dǎo)組;第四次全國(guó)結(jié)核病流行病學(xué)抽樣調(diào)查報(bào)告[J];中華結(jié)核和呼吸雜志;2002年01期
3 王家祥;王利;范應(yīng)中;劉秋亮;張蛟;余捷凱;鄭樹;;基于支持向量機(jī)的血清蛋白質(zhì)指紋圖譜模型在甲狀腺癌診斷中的應(yīng)用研究[J];中華醫(yī)學(xué)雜志;2006年14期
4 蘇玉亮;張寰;曲金力;錢碧云;;EGFR突變相關(guān)miR-335靶基因預(yù)測(cè)及其功能的生物信息學(xué)分析[J];中國(guó)腫瘤臨床;2013年10期
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