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采用斑点印迹酶联免疫吸附测定法检测丙型病毒性肝炎合并血吸虫病患者的血清层粘连蛋白。

Serum laminin assayed by Slot-Blot-ELISA in patients with combined viral hepatitis C and schistosomiasis.

作者信息

El-Masry Samir, Lotfy Mahmoud, El-Shahat Mohamed, Badra Gamal

机构信息

Molecular and Cellular Biology Department, Genetic Engineering and Biotechnology Research Institute, Minufiya University, Sadat City, P.O. 79, Minufiya, Egypt.

出版信息

Clin Biochem. 2006 Jun;39(6):652-7. doi: 10.1016/j.clinbiochem.2005.12.008. Epub 2006 Feb 20.

Abstract

OBJECTIVES

Hepatic schistosomiasis and chronic hepatitis C virus (HCV) are the most prevalent agents causing hepatic fibrosis in humans. Laminin (LA) has been related to liver fibrosis and subsequent development of portal hypertension in chronic liver disease. There are no available data describing the pattern of laminin in combined HCV and schistosoma-infected patients, thus the rationale of this study was to assess the serum LA as an index of liver fibrosis in patients with schistosomiasis and/or chronic viral hepatitis C and to evaluate a developed Slot-Blot Enzyme-Linked Immunosorbant Assay (Slot-Blot-ELISA) as a method of estimation.

DESIGNS AND METHODS

This study included four groups: group I included 34 patients with schistosomiasis, group II included 58 patients infected with HCV, group III included 68 patients with combined chronic viral hepatitis C and schistosomiasis and group IV included 50 healthy individuals who served as a control group. Serum LA was measured in the different groups quantitatively by ELISA and semi-quantitatively by Slot-Blot-ELISA.

RESULTS

Significantly higher serum LA concentrations measured by ELISA were found in patients with combined chronic viral hepatitis C and schistosomiasis than in patients with either chronic HCV (P = 0.005) or schistosomiasis (P < 0.001) alone. Serum LA was significantly higher in the patient groups than the control group (P < 0.001). Serum LA concentration was positively correlated with fibrosis grading scores. Semi-quantitative results of serum LA using the developed SB-ELISA were found to have approximately the same power of ELISA results in different groups. The overall sensitivity, specificity, positive predictive value, negative predictive value and efficiency of ELISA for estimation of serum LA were 85.6%, 84.0%, 94.5%, 64.6% and 90%, respectively and for SB-ELISA were 87.5%, 82.0%, 94%, 67.2% and 88%, respectively.

CONCLUSIONS

Serum LA was significantly increased in patients coinfected with HCV and Schistosoma mansoni. The newly developed Slot-Blot-ELISA is a simple, rapid and highly sensitive assay for detection of LA in hepatic fibrosis. Moreover, all steps were performed at room temperature without the need to use expensive equipment, and this may enhance the application of this assay in screening programs. Further study is warranted for confirmation of SB-ELISA reproducibility in a large population.

摘要

目的

肝血吸虫病和慢性丙型肝炎病毒(HCV)是导致人类肝纤维化的最常见病因。层粘连蛋白(LA)与肝纤维化及慢性肝病中门静脉高压的后续发展有关。目前尚无关于HCV和血吸虫合并感染患者中层粘连蛋白模式的数据,因此本研究的目的是评估血清LA作为血吸虫病和/或慢性丙型病毒性肝炎患者肝纤维化指标,并评估一种改良的斑点印迹酶联免疫吸附测定法(Slot-Blot-ELISA)作为一种评估方法。

设计与方法

本研究包括四组:第一组包括34例血吸虫病患者,第二组包括58例HCV感染患者,第三组包括68例慢性丙型病毒性肝炎合并血吸虫病患者,第四组包括50名健康个体作为对照组。通过ELISA对不同组定量检测血清LA,通过Slot-Blot-ELISA进行半定量检测。

结果

ELISA检测显示,慢性丙型病毒性肝炎合并血吸虫病患者的血清LA浓度显著高于单独患有慢性HCV(P = 0.005)或血吸虫病(P < 0.001)的患者。患者组血清LA显著高于对照组(P < 0.001)。血清LA浓度与纤维化分级评分呈正相关。使用改良的SB-ELISA对血清LA进行半定量检测,其结果在不同组中与ELISA结果具有大致相同的效力。ELISA评估血清LA的总体敏感性、特异性、阳性预测值、阴性预测值和效率分别为85.6%、84.0%、94.5%、64.6%和90%,SB-ELISA分别为87.5%、82.0%、94%、67.2%和88%。

结论

HCV和曼氏血吸虫合并感染患者的血清LA显著升高。新开发的Slot-Blot-ELISA是一种用于检测肝纤维化中LA的简单、快速且高度灵敏的检测方法。此外,所有步骤均在室温下进行,无需使用昂贵设备,这可能会提高该检测方法在筛查项目中的应用。需要进一步研究以证实SB-ELISA在大量人群中的可重复性。

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