GBP5、KLF2基因在潜伏结核感染人群中表达研究

发布时间：2018-03-30 07:31

本文选题：潜伏结核感染　切入点：GBP5　出处：《青岛大学》2017年硕士论文

【摘要】：目的:探讨GBP5和KLF2这两个目的基因在潜伏结核感染人群中的表达情况,并同时与它们在涂阳结核病患者及健康人群中表达情况进行比较。分析两种目的基因在三组人群中相对表达量的差异,确定潜伏结核感染(LTBI)人群中将来有可能发展成为结核病患者的表达阈值。为LTBI高危人群的筛选找到合适的标志物,以便对这组人群进行精准的化学预防,控制结核病的发生。方法:在一家综合医院呼吸科和一家结核病专科医院两家单位,共收集180例受试者,分为三组,以痰涂片是否找到结核分枝杆菌(MTB))和γ-干扰素释放试验QFT结果为标准,分别为涂阳结核病患者组60例,潜伏结核感染者组(QFT阳性)60例,健康人群组(QFT阴性)60例,对符合条件的受试者,每人抽取外周全血4ml,并给予肝素锂抗凝处理。然后在两小时内,置于离心机内,分离T淋巴细胞,提取RNA。然后通过反转录酶作用,合成c DNA。再以c DNA为模板,通过前期合成的引物引导,合成目的基因片段。对合成的目的基因片段采用RT-PCR技术,进行基因扩增,通过定时定量仪器,测得两种目的基因的CT值。用2-△△CT法,对目的基因进行差值校正,求得两种目的基因GBP5、KLF2在三组受试者中的相对表达量。通过SPSS18.0统计分析软件对GBP5、KLF2在三组人群的相对表达量数值进行统计学分析,观察差异是否有统计学意义。如果统计分析有效,则进一步进行均值的比较,并分别确定两种目的基因的表达阈值。同时对可能存在的其他情况进行分析比较。结果:1、三组受试者中,GBP5相对表达量差异有统计学意义(p㩳0.05),GBP5相对表达量,在涂阳结核病患者组中表达最高,在潜伏结核感染人群组中次之,在健康人群组中表达最低。2、三组受试者中,KLF2相对表达量差异有统计学意义(p㩳0.05),KLF2相对表达量,在涂阳结核病患者组中表达最高,在潜伏结核感染者中次之,在健康人群组组中表达最低。3、对于潜伏结核感染人群中远期进展为结核病可能性高的群体,上述两个目的基因表达阈值的界定:GBP5相对表达量在1.17~1.25范围内,未来发病风险高,是精准预防的对象。KLF2相对表达量在1.11~1.21范围内,未来发病风险高,是精准预防的对象。4、GBP5、KLF2可以区别潜伏结核感染与活动性结核病。结论:1、GBP5在三组受试者中相对表达量不同,GBP5可以用来筛选潜伏结核感染人群中远期进展为结核病可能性高的群体。2、KLF2在三组受试者中相对表达量不同,KLF2可以用来筛选潜伏结核感染人群远期进展为结核病可能性高的群体。GBP5、KLF2可以为LTBI高危人群精准化学预防提供诊断依据。3、GBP5、KLF2可以用于活动性结核病与潜伏结核感染的鉴别性诊断。4、GBP5、KLF2有望成为结核病基因诊断的目的基因。
[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.

【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R52

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