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2013年在伊朗拉夫桑詹市献血者中运用实时荧光定量聚合酶链反应和酶联免疫吸附测定技术对急性和潜伏性弓形虫病进行分子和血清学检测

Molecular and serological detection of acute and latent toxoplasmosis using real-time PCR and ELISA techniques in blood donors of rafsanjan city, iran, 2013.

作者信息

Zainodini Nahid, Zare-Bidaki Mohammad, Abdollahi Seyyed Hossein, Afrooz Mohammadreza, Ziaali Naser, Ebrahimian Mohammad, Kazemi Arababadi Mohammad

机构信息

Immunology of Infectious Diseases Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran.

Molecular Medicine Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran.

出版信息

Iran J Parasitol. 2014 Sep;9(3):336-41.

Abstract

BACKGROUND

The differentiation between acute and latent forms of the Toxoplasma gondii (T. gondii) infection is still considered as a complicated issue. This study was aimed to elucidate the status of infection in the blood donors and the probable importance of blood transfusion in the transmission of the infection through detecting both immunological and genetic markers of acute and latent infection.

METHODS

Totally 235 blood samples from blood donors were collected. The levels of anti-T. gondii IgG and IgM antibodies were examined by specific ELISA kits. cDNA were synthesized from total extracted mRNA molecules from the serum samples and SAG1 gene, specific for tachyzoite form, were amplified using Real-Time PCR technique. Demographic information of study subjects including their gender, age, job, and habitat were recorded.

RESULTS

Out of 235 serum samples, 80 (34.04%) and 4 (1.71%) were positive regarding anti-T. gondii IgG and IgM antibodies, respectively. Real-Time PCR results showed that 14 out of 200 (6.97%) of blood donor had mRNA molecules of SAG1 gene. The positive results of Real-Time PCR of SAG1 in female gender and housekeepers were significantly higher than those of male gender and other job categories.

CONCLUSION

The prevalence of chronic and acute infection is high in Iranian blood donors. Additionally, evaluation of antibodies could not be reliable, because several donors negative for anti-T. gondii IgM antibodies had detectable SAG1 mRNA molecules. Hence, it seems that molecular diagnostic tests are essential to detect acute infections.

摘要

背景

弓形虫感染的急性和潜伏形式之间的区分仍被视为一个复杂的问题。本研究旨在通过检测急性和潜伏感染的免疫和基因标志物,阐明献血者的感染状况以及输血在感染传播中的可能重要性。

方法

共收集了235份来自献血者的血样。使用特定的ELISA试剂盒检测抗弓形虫IgG和IgM抗体水平。从血清样本中提取的总mRNA分子合成cDNA,并使用实时PCR技术扩增速殖子形式特异的SAG1基因。记录研究对象的人口统计学信息,包括他们的性别、年龄、职业和居住地。

结果

在235份血清样本中,分别有80份(34.04%)和4份(1.71%)抗弓形虫IgG和IgM抗体呈阳性。实时PCR结果显示,200名献血者中有14名(6.97%)具有SAG1基因的mRNA分子。女性和家政人员中SAG1实时PCR的阳性结果显著高于男性和其他职业类别。

结论

伊朗献血者中慢性和急性感染的患病率较高。此外,抗体评估可能不可靠,因为一些抗弓形虫IgM抗体阴性的献血者具有可检测到的SAG1 mRNA分子。因此,似乎分子诊断测试对于检测急性感染至关重要。

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