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从沙特阿拉伯阿西尔地区的沙特孕妇采集的血液样本中通过聚合酶链反应检测弓形虫DNA。

Detection of Toxoplasma gondii DNA by PCR in blood samples collected from pregnant Saudi women from the Aseer region, Saudi Arabia.

作者信息

Bin Dajem Saad M, Almushait Mona A

机构信息

Biology Department, Faculty of Medicine, King Khalid University College of Science, Abha, Saudi Arabia.

出版信息

Ann Saudi Med. 2012 Sep-Oct;32(5):507-12. doi: 10.5144/0256-4947.2012.14.7.1200.

Abstract

BACKGROUND AND OBJECTIVES

Toxoplasmosis, caused by Toxoplasma gondii, is diagnosed mainly by serological methods that are hindered by insufficient sensitivity. When it fails, it becomes necessary to rely on either direct detection of the parasite or DNA detection by polymerase chain reaction (PCR). We aimed to establish molecular tools for toxoplasmosis research in the country by using PCR targeting the B1 gene and compare it with ELISA results.

DESIGN AND SETTING

Conducted at the College of Science, King Khalid University, Abha, Saudi Arabia between January 2009 and April 2010 on Saudi pregnant women attending three major hospitals in the Aseer region.

PATIENTS AND METHODS

Peripheral blood samples (n=137) were collected from patients. DNA was extracted and the B1 T gondii gene was amplified by PCR. The amplicons were visualized and sequenced, and the results were analyzed. For comparison, sera were tested for anti-T gondii IgG and IgM by enzyme-linked immunosorbent assay (ELISA).

RESULTS

Of the 137 samples tested, the B1 gene could be amplified in 56 cases (41%) by PCR. DNA sequencing confirmed these results. IgM-ELISA assay detected 9 (6.5%) of these cases. The results of immunoglobulin G detection were positive in 53 (38.6%) of the patients.

CONCLUSION

The present study showed the need for PCR as a confirmatory assay in addition to serological assays to detect recent infection. We recommend national implementation of these molecular diagnostic tools.

摘要

背景与目的

由刚地弓形虫引起的弓形虫病,主要通过血清学方法诊断,但该方法存在灵敏度不足的问题。当血清学方法失效时,就需要依靠直接检测寄生虫或通过聚合酶链反应(PCR)检测DNA。我们旨在通过靶向B1基因的PCR技术建立该国弓形虫病研究的分子工具,并将其与酶联免疫吸附测定(ELISA)结果进行比较。

设计与地点

2009年1月至2010年4月在沙特阿拉伯阿卜哈市的哈立德国王大学理学院对阿西尔地区三家主要医院的沙特孕妇进行了研究。

患者与方法

采集患者外周血样本(n = 137)。提取DNA,通过PCR扩增刚地弓形虫B1基因。对扩增产物进行可视化和测序,并分析结果。为作比较,采用酶联免疫吸附测定(ELISA)检测血清中的抗刚地弓形虫IgG和IgM。

结果

在检测的137个样本中,PCR可扩增出B1基因的有56例(41%)。DNA测序证实了这些结果。IgM-ELISA检测出其中9例(6.5%)。免疫球蛋白G检测结果显示53例(38.6%)患者呈阳性。

结论

本研究表明,除血清学检测外,还需要PCR作为确认检测方法来检测近期感染。我们建议在全国范围内应用这些分子诊断工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa05/6080984/5d07048181d6/asm-5-507f1.jpg

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