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使用TaqMan荧光探针通过实时聚合酶链反应检测免疫功能低下患者和孕妇感染的急性期和慢性期。

Detection of in Acute and Chronic Phases of Infection in Immunocompromised Patients and Pregnant Women with Real-time PCR Assay Using TaqMan Fluorescent Probe.

作者信息

Mousavi Parisa, Mirhendi Hossein, Mohebali Mehdi, Shojaee Saeedeh, Keshavarz Valian Hossein, Fallahi Shirzad, Mamishi Setareh

机构信息

Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

Dept. of Medical Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Iran J Parasitol. 2018 Jul-Sep;13(3):373-381.

Abstract

BACKGROUND

, cause severe medical complications in infants and immune-compromised individuals. As using early, sensitive and rapid technique has major in diagnosis of toxoplasmosis, the present study was aimed to detect parasite by using from repetitive element (RE) and B1genes, in blood samples of seropositive immuno-compromised patients and pregnant women.

METHODS

A total of 110 peripheral blood samples were collected from seropositive cases with anti- antibodies, including immunocompromised patients and pregnant women. DNA was extracted by a commercial kit and subjected to TaqMan probe-based real-time PCR assay by using primers and probes specific for RE and B1 genes, separately. The data were analyzed by Kappa test and SPSS-22 software.

RESULTS

In the pregnant women, 17 (68%) and 14 (56%) samples from 25 IgM+/ IgG+ cases and, 7 (25%) and 6 (21.4%) samples from 28 IgG+/IgM- cases were positive by RE and B1 real time PCR, respectively. Likewise, in immunocompromised group, 20 (66.6%) and 17 (56.6%) samples from 30 IgM+/ IgG+ cases and 2 (7.4%) and 2 (7.4%) samples from 27 IgG+/ IgM- cases were positive by RE and B1 real time PCR, respectively.

CONCLUSION

Probe-based real time PCR assay is a quantitative approach for early diagnosis of infection in clinical samples. Moreover, this method can be more appropriate in diagnosis of acute and reactivated toxoplasmosis. In addition our results indicated that RE gene is more sensitive than B1 gene.

摘要

背景

[原文此处不完整]在婴儿和免疫功能低下个体中会引发严重的医学并发症。由于采用早期、灵敏且快速的技术对弓形虫病诊断至关重要,本研究旨在通过使用重复元件(RE)和B1基因,检测血清学阳性的免疫功能低下患者和孕妇血液样本中的寄生虫。

方法

从具有抗[抗体名称原文此处缺失]抗体的血清学阳性病例中总共采集了110份外周血样本,包括免疫功能低下患者和孕妇。使用商业试剂盒提取DNA,并分别使用针对RE和B1基因的引物和探针进行基于TaqMan探针的实时PCR检测。数据通过Kappa检验和SPSS - 22软件进行分析。

结果

在孕妇中,25例IgM + / IgG +病例的17份(68%)和14份(56%)样本,以及28例IgG + / IgM -病例的7份(25%)和6份(21.4%)样本通过RE和B1实时PCR分别呈阳性。同样,在免疫功能低下组中,30例IgM + / IgG +病例的20份(66.6%)和17份(56.6%)样本,以及27例IgG + / IgM -病例的2份(7.4%)和2份(7.4%)样本通过RE和B1实时PCR分别呈阳性。

结论

基于探针的实时PCR检测是临床样本中弓形虫感染早期诊断的定量方法。此外,该方法在急性和再激活弓形虫病的诊断中可能更适用。此外,我们的结果表明RE基因比B1基因更灵敏。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbb9/6243173/533638c48a39/IJPA-13-373-g001.jpg

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