梅毒患者血液47KDa、15KDa膜免疫原基因的测定

发布时间：2018-06-10 00:52

本文选题：梅毒螺旋体 + 巢式聚合酶链反应(PCR)　；参考：《兰州大学》2012年硕士论文

【摘要】：背景：梅毒(sypilis)是由梅毒螺旋体(Treponema pallidum, TP)(又称苍白螺旋体)引起的一种全身性慢性传染性疾病,可侵犯人体所有器官,其病程较长,临床表现非常复杂,并可通过胎盘传播,引起胎儿先天性梅毒或妊娠不良,危害下一代健康。梅毒的广泛流行和传播已成为世界各国的公共卫生问题,因此梅毒的规范化治疗也受到高度重视。治疗梅毒的原则是诊断明确、尽早治疗、剂量充足、疗程规则、治疗后随访观察、配偶或性伴同时接受检查和治疗、治疗期间禁止性交,力争达到临床和血清学均治愈。然而诊断梅毒时,在某些情况下,梅毒血清学试验会出现假阳性、假阴性结果,给梅毒的诊治带来很大的困扰。在有关梅毒的各类研究中,临床免疫学是其中的热点。近年来,随着基因克隆技术的不断发展,梅毒全基因序列得到确定,其基因编码的众多抗原中,TpN15、TpN17、TpN44.5、TpN47等外膜蛋白均可用于TP的检测。目前国内外尚没有通过检测TpN47、TpN15这两种膜免疫原基因来诊断梅毒的报道,本实验运用巢式PCR检测梅毒患者血液中47KDa、15KDa这两种膜免疫原基因以排除梅毒血清学假阳性。目的：用巢式聚合酶链反应(PCR)扩增全血中梅毒螺旋体(TP)的47KDa、15KDa的膜免疫原基因,以排除梅毒血清学试验假阳性。方法：应用巢式PCR和常规PCR分别扩增90例疑似梅毒病人全血中的TpN15、TpN47,并与血清学方法比较。结果：通过检测标本,并与血清学方法比较,巢式PCR检测TP的敏感性和特异性分别为93.3%和96.7%,两者符合率为96.7%;两种PCR方法检测结果差异有显著性(P0.01),巢式PCR敏感性高于常规PCR。结论：巢式PCR检测全血中TP较常规PCR灵敏,通过检测47KDa、15KDa的膜免疫原基因,有助于排除梅毒血清学试验假阳性,提高梅毒的诊断准确率。
[Abstract]:Background: syphilis is a chronic systemic infectious disease caused by Treponema pallidumi (TPN) of Treponema pallium, which can invade all organs of human body. The course of disease is long, the clinical manifestation is very complicated, and it can be transmitted through placenta. Causes fetal congenital syphilis or poor pregnancy, harmful to the health of the next generation. The widespread prevalence and spread of syphilis has become a public health problem all over the world, so the standardized treatment of syphilis has also been attached great importance. The principles of treatment for syphilis are clear diagnosis, early treatment, adequate dosage, regular course of treatment, follow-up observation after treatment, examination and treatment of spouse or sexual partner at the same time, prohibition of sexual intercourse during treatment, and strive to achieve both clinical and serological cure. However, the diagnosis of syphilis, in some cases, syphilis serological test will appear false positive, false negative results, the diagnosis and treatment of syphilis brings great trouble. Clinical immunology is one of the hotspots of syphilis research. In recent years, with the development of gene cloning technology, the whole gene sequence of syphilis has been determined. The outer membrane proteins such as TpN15, TpN17, TpN44.5 and TpN47 can be used for the detection of TP. At present, there are no reports at home and abroad on the diagnosis of syphilis by detecting TpN47 and TpN15, two kinds of membrane immunogen genes. In this study, we used nested polymerase chain reaction (NPCR) to detect the membrane immunogen genes of 47 K Da1 15 KDa in the blood of syphilis patients in order to exclude the false positive serology of syphilis. Objective: to amplify the membrane immunogen gene of 47 K Da1 15 KDa in the whole blood by nested polymerase chain reaction (PCR). Methods: TpN15 TpN47 was amplified by nested PCR and routine PCR in 90 patients with suspected syphilis, and compared with the serological method. Results: the samples were detected and compared with the serological method. The sensitivity and specificity of nested PCR for TP detection were 93.3% and 96.7% respectively, the coincidence rate was 96.70.There was significant difference between the two PCR methods, and the sensitivity of nested PCR was higher than that of conventional PCR.Conclusion: nested PCR is more sensitive than conventional PCR in detecting TP in whole blood. The detection of the membrane immunogen gene of 47K Dahe 15KDa is helpful to eliminate the false positive of syphilis serological test and to improve the diagnostic accuracy of syphilis.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R759.1

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