本文選題：潛伏結(jié)核感染　切入點：GBP5　出處：《青島大學》2017年碩士論文

【摘要】：目的:探討GBP5和KLF2這兩個目的基因在潛伏結(jié)核感染人群中的表達情況,并同時與它們在涂陽結(jié)核病患者及健康人群中表達情況進行比較。分析兩種目的基因在三組人群中相對表達量的差異,確定潛伏結(jié)核感染(LTBI)人群中將來有可能發(fā)展成為結(jié)核病患者的表達閾值。為LTBI高危人群的篩選找到合適的標志物,以便對這組人群進行精準的化學預防,控制結(jié)核病的發(fā)生。方法:在一家綜合醫(yī)院呼吸科和一家結(jié)核病專科醫(yī)院兩家單位,共收集180例受試者,分為三組,以痰涂片是否找到結(jié)核分枝桿菌(MTB))和γ-干擾素釋放試驗QFT結(jié)果為標準,分別為涂陽結(jié)核病患者組60例,潛伏結(jié)核感染者組(QFT陽性)60例,健康人群組(QFT陰性)60例,對符合條件的受試者,每人抽取外周全血4ml,并給予肝素鋰抗凝處理。然后在兩小時內(nèi),置于離心機內(nèi),分離T淋巴細胞,提取RNA。然后通過反轉(zhuǎn)錄酶作用,合成c DNA。再以c DNA為模板,通過前期合成的引物引導,合成目的基因片段。對合成的目的基因片段采用RT-PCR技術(shù),進行基因擴增,通過定時定量儀器,測得兩種目的基因的CT值。用2-△△CT法,對目的基因進行差值校正,求得兩種目的基因GBP5、KLF2在三組受試者中的相對表達量。通過SPSS18.0統(tǒng)計分析軟件對GBP5、KLF2在三組人群的相對表達量數(shù)值進行統(tǒng)計學分析,觀察差異是否有統(tǒng)計學意義。如果統(tǒng)計分析有效,則進一步進行均值的比較,并分別確定兩種目的基因的表達閾值。同時對可能存在的其他情況進行分析比較。結(jié)果:1、三組受試者中,GBP5相對表達量差異有統(tǒng)計學意義(p㩳0.05),GBP5相對表達量,在涂陽結(jié)核病患者組中表達最高,在潛伏結(jié)核感染人群組中次之,在健康人群組中表達最低。2、三組受試者中,KLF2相對表達量差異有統(tǒng)計學意義(p㩳0.05),KLF2相對表達量,在涂陽結(jié)核病患者組中表達最高,在潛伏結(jié)核感染者中次之,在健康人群組組中表達最低。3、對于潛伏結(jié)核感染人群中遠期進展為結(jié)核病可能性高的群體,上述兩個目的基因表達閾值的界定:GBP5相對表達量在1.17~1.25范圍內(nèi),未來發(fā)病風險高,是精準預防的對象。KLF2相對表達量在1.11~1.21范圍內(nèi),未來發(fā)病風險高,是精準預防的對象。4、GBP5、KLF2可以區(qū)別潛伏結(jié)核感染與活動性結(jié)核病。結(jié)論:1、GBP5在三組受試者中相對表達量不同,GBP5可以用來篩選潛伏結(jié)核感染人群中遠期進展為結(jié)核病可能性高的群體。2、KLF2在三組受試者中相對表達量不同,KLF2可以用來篩選潛伏結(jié)核感染人群遠期進展為結(jié)核病可能性高的群體。GBP5、KLF2可以為LTBI高危人群精準化學預防提供診斷依據(jù)。3、GBP5、KLF2可以用于活動性結(jié)核病與潛伏結(jié)核感染的鑒別性診斷。4、GBP5、KLF2有望成為結(jié)核病基因診斷的目的基因。
[Abstract]:Objective: To investigate the GBP5 and KLF2 two gene expression in the latent tuberculosis infection in the crowd, and at the same time and their expression in smear positive tuberculosis patients and healthy people. The comparison of two kinds of relative gene expression differences among three groups, determine the latent tuberculosis infection (LTBI) in the crowd the future may become the expression threshold of TB patients. Screening for LTBI in high-risk population to find suitable markers, so that accurate chemical prevention of this population, control of tuberculosis. Methods: in the Department of respiration of a comprehensive hospital and a tuberculosis specialist hospital two units, a total of 180 subjects, divided into three groups, to find whether sputum smear of Mycobacterium tuberculosis (MTB)) and interferon gamma release test results of QFT as the standard, were smear positive TB patients in group 60 cases, latent tuberculosis infection group (QFT Yang Of the 60 cases) healthy group (QFT negative) 60 cases of eligible subjects, each from peripheral blood 4ml, and heparin lithium anticoagulant treatment. Then in two hours, placed in the centrifuge separation, extraction of RNA. and T lymphocytes, by reverse transcriptase, synthesis of C DNA. taking C DNA as the template, through the pre synthesized primer guide, synthetic gene fragments of target gene fragment synthesis using RT-PCR technology, gene amplification, through quantitative timing instrument, measured two gene CT value. Using 2- Delta CT method, in order to gene difference correction, obtained two a GBP5 gene, KLF2 relative expression of subjects in the three groups. The analysis software of GBP5 by SPSS18.0 statistics, KLF2 in the relative expression of the three groups were statistically analyzed value to observe the difference is statistically significant. If the effective statistical analysis, is a Comparison of mean step, and determine the expression of target genes of the two threshold. At the same time for other possible conditions were analyzed and compared. Results: 1. Three groups of subjects, the relative expression of GBP5 was statistically significant difference (P? 0.05), the relative expression of GBP5 in sputum smear positive TB patients the highest expression, in the latent tuberculosis infection group in the second, the lowest.2 expression in healthy people group, the three groups of subjects, the relative expression of KLF2 was statistically significant difference (P? 0.05), the relative expression of KLF2 reached the highest in sputum smear positive TB patients, in latent tuberculosis infection in the second, in the.3 group was the lowest in the healthy population, for latent tuberculosis infection in the crowd for the long-term high possibility of tuberculosis group, the two gene expression threshold definition: the relative expression of 1.17~1.25 in the range of GBP5, the future risk is high, fine Quasi relative expression in the range of 1.11~1.21 to prevent object.KLF2 future risk prevention is high, precise object.4, GBP5, KLF2 can distinguish latent tuberculosis infection and active tuberculosis. Conclusion: 1. GBP5 in the three groups of subjects in the relative expression of different GBP5 can be used for screening of latent tuberculosis infection progress the crowd forward for tuberculosis with high possibility of group.2, KLF2 in the three groups of subjects in the relative expression of different KLF2 can be used for screening of latent tuberculosis infection in long-term progress to higher risk of TB groups.GBP5, KLF2 for LTBI high risk population accurate chemical prevention and provide a basis of diagnosis of.3, GBP5, KLF2 can be used for activity differential diagnosis of tuberculosis and latent tuberculosis infection in.4, GBP5, KLF2 is expected to become a target gene for diagnosis of tuberculosis.

【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R52

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