| 洪万东,朱启槐,陈向荣.幽门螺杆菌粪便抗原免疫卡诊断幽门螺杆菌感染的Meta分析[J].Chinese journal of Epidemiology,2008,29(1):71-74 |
| 幽门螺杆菌粪便抗原免疫卡诊断幽门螺杆菌感染的Meta分析 |
| Study on the value of Helicobacter pyiori(H.pylori)stool antigen ImmunoCard STAT in the diagnosis of H.pylori infection:a Meta-analysis |
| Received:August 30, 2007 Revised:June 28, 2012 |
| DOI： |
| KeyWord: 幽门螺杆菌 粪便抗原 诊断试验 Meta分析 |
| English Key Word: Hantavirus Nueleocapsid protein Recombinant expression IgM antibody Capture enzyme-linked immunosorbent assay |
| FundProject:浙江省科技厅重点科研资助项目(2005C23014);浙江省医药卫生科学研究基金资助项目(2006B020) |
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| Abstract: |
| 目的 评价幽门螺杆菌粪便抗原免疫卡诊断幽门螺杆菌感染的精确性.方法 计算机检索Medline(1966-2007年4月)、EMbase(1985-2007年4月)和中国期刊全文数据库(1994-2007年)有关幽门螺杆菌粪便抗原免疫卡诊断幽门螺杆菌感染的临床试验，按Cochrane协作网推荐的方法进行Meta分析.结果 共11篇文献被纳入评价.合并敏感性和合并特异性分别为0.93(95%CI:0.91～0.94)、0.93(95%CI:0.90～0.95)，合并阳性似然比和合并阴性似然比分别为12.01(95%CI:8.90～16.19)、0.08(95%C/:0.07～0.11).合并诊断优势比为160.14(95%CI:100.43～255.34).SROC曲线下面积为0.974±0.005.结论 幽门螺杆菌粪便抗原免疫卡是一种精确无创的诊断成年人幽门螺杆菌感染方法。 |
| English Abstract: |
| Objective To clone the gene encoding nucleocapsid protein(NP) of hantavirus strain Z10(HV-Z10),to construct its prokaryotic expression system as well as to establish a rNP-IgM direct capture ELISA based on HRP-labeled recombinant NP(rNP),in order to detect serum samples of patients suffering from hemorrhagic fever with renal syndrome(HFRS) and to evaluate the effects of detection. Methods Gene encoding NP of strain HV-Z10 was amplified by PCR and then its prokaryotic expression system pET28a-Z10N-E,coli BL21DE3 was constructed,using routine genetic engineering method.SDS- PAGE was applied to measure the expression of rNP and ion-exchange plus Ni-NTA-affinity chromatography was performed to purify the recombinant product.Western blot assay was used to determine the specific immuno-reactivity of rNP while HRP-labeled rNP-IgM direct capture ELISA was established to detect the serum samples from 95 cases of confirmed HFRS patients.The detection effect was compared with that by routine HV-IgM indirect capture ELISA method.Results pET28a-Z10N-E,coli BL21DE3 was able to express rNP with high efficiency.The purified rNP only showed a single protein fragment in the gel after SDS-PAGE.HV-IgG could efficiently recognize rNP and hybridize with the recombinant protein.94.73% (90/95) of HFRS patients' serum samples were positively confirmed by rNP- IgM direct capture ELISA,while a positive rate of 92.63% (88/95) in the same samples was confirmed by HV-IgM indirect capture ELISA.The distributions of A450) values of the serum samples detected by the two IgM capture ELISAs as well as the changes of the A450) mean values from several serum samples with different dilutions were similar.Conclusion We successfully constructed a high efficient prokaryotic expression system of NP encoding gene of hantavirus strain HV-Z10.The rNP-IgM direct capture ELISA that established in this study could be used as a new serological test for HFRS diagnosis because of its simplicity,safety,with high sensitivity and specificity. |
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