摘要
目的研究肺炎支原体(MP)特异性抗原免疫反应性及检测合肥地区138份儿童血清样本感染MP的情况,并初步分析其危险因素。方法构建含r P1-513基因的克隆和表达质粒分别命名为T-r P1-513和PQE-80L-r P1-513。诱导PQE-80L-r P1-513重组菌蛋白表达。Western blot法检测其免疫反应性和ELISA法筛选血清样本感染MP的阳性标本。结果 Western blot法检测约30 ku的r P1-513蛋白有免疫反应性。ELISA法筛选出34份阳性血清,同时用商业MP ELISA试剂盒检测出24份阳性血清,两种P1蛋白作为抗原ELISA检测法检出率分别为24.6%和17.4%,且两个检出率差异有统计学意义(P<0.05)。MP感染与C-反应蛋白有显著相关性,但与性别、年龄、白细胞无相关性。结论初步证明r P1-513蛋白具有免疫反应性,且以此蛋白构建的ELISA法有较高的灵敏度和特异性,为临床诊断MP提供一定的帮助。
Objective To study the specific antigen immune reactivity,detect its infection situation in 138 children serum samples in Anhui of Mycoplasma pneumoniae （MP）,and analyze preliminarily its risk factors.Methods The recombinant cloning and expression plasmids with the target gene were named as T-rP1-513 and PQE-80L-rP1-513.The protein of PQE-80L-rP1-513 recombinant bacteria was induced to express.In this study,the protein immune reactivity was detected by Western blot and antibodies to the P1 protein of MP were tested by ELISA in serum.Results The rP1-513 protein with approximately 30 ku had immune reactivity by Western blot.34 positive specimens were screened by ELISA.At the same time,24 positive specimens were found by the commercial MP ELISA kit.Their detection rates were 24.6％ and 17.4％ respectively,and there was a statistically significant difference（P ＜ 0.05）.MP infection and C-reactive protein had a significant correlation except gender,age and white blood cells.Conclusion It is preliminarily confirmed that ELISA method with rP1-513 protein has higher sensitivity and specificity,and can provide certain help for clinical diagnosis of MP.
出处
《安徽医科大学学报》
CAS
北大核心
2014年第12期1709-1713,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81271837)
安徽省教育厅自然科学基金(编号:KJ2012A161)
安徽医科大学博士科研经费资助项目(编号:XJ200914)
