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针对克隆的弓形虫寄生虫的两种聚合酶链反应方案的性能

Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites.

作者信息

Hanafy Nihal A, Badr Mohamed S, Nasr Ghada M

机构信息

Department of Parasitology, Faculty of Medicine, Cairo University, Cairo, Egypt.

Department of Microbiology, International Medical School, Management and Science University, Shah Alam, Malaysia.

出版信息

Open Access Maced J Med Sci. 2018 Sep 22;6(9):1577-1580. doi: 10.3889/oamjms.2018.400. eCollection 2018 Sep 25.

Abstract

BACKGROUND

is a common parasitic infection of humans. Infection is usually mild. Serious complications can occur in pregnant and immunocompromised patients.

AIM

The present study aims to investigate the performance of 2 different PCR protocols; real-time quantitative molecular assays (qPCR) and conventional molecular assays (cPCR), using 2 different sets of primers and by using cloned purified Toxoplasma genomic substances to be evaluated as reference samples.

METHODS

The target DNA was provided in 8 different quantities.

RESULTS

Amplification failure was reported only with the cPCR in samples of low concentrations using both primer sets. Quantitative PCR detected the 8 different dilutions of the purified using the 2 sets of primers while cPCR was sensitive to detect only 6 different dilutions.

CONCLUSION

Generally real-time quantitative molecular assays, is easy to use method compared to conventional PCR assay and produces more reliable results within only one hour time but still the possible application of qPCRs in routine diagnosis necessitates analysis of a large number of clinical samples in further studies to make the proper choice.

摘要

背景

是一种常见的人类寄生虫感染。感染通常较轻。孕妇和免疫功能低下的患者可能会出现严重并发症。

目的

本研究旨在使用两组不同的引物,并通过使用克隆纯化的弓形虫基因组物质作为参考样本,来研究两种不同的PCR方案;实时定量分子检测(qPCR)和传统分子检测(cPCR)的性能。

方法

以8种不同的量提供目标DNA。

结果

使用两组引物时,仅在低浓度样本的cPCR中报告了扩增失败。定量PCR使用两组引物检测到了纯化的8种不同稀释度,而cPCR仅能敏感地检测到6种不同稀释度。

结论

一般来说,与传统PCR检测相比,实时定量分子检测是一种易于使用的方法,并且在仅一小时的时间内就能产生更可靠的结果,但qPCR在常规诊断中的可能应用仍需要在进一步研究中分析大量临床样本,以便做出正确选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0dae/6182548/9e467096f4be/OAMJMS-6-1577-g001.jpg

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