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通过磁性分离、厚血膜显微镜检查和逆转录聚合酶链反应检测恶性疟原虫配子体的敏感性比较

A comparison of the sensitivities of detection of Plasmodium falciparum gametocytes by magnetic fractionation, thick blood film microscopy, and RT-PCR.

作者信息

Karl Stephan, Davis Timothy M E, St-Pierre Tim G

机构信息

School of Physics, M013, The University of Western Australia, 35 Stirling Highway, Crawley WA 6009, Australia.

出版信息

Malar J. 2009 May 11;8:98. doi: 10.1186/1475-2875-8-98.

Abstract

BACKGROUND

The magnetic properties of Plasmodium-infected erythrocytes have been exploited for different clinical and research purposes. A recent study in a rural clinical setting in Papua New Guinea has demonstrated that Plasmodium falciparum gametocyte detection is facilitated by magnetic deposition microscopy but no study has yet determined the relative sensitivity and limit of detection of a magnetic fractionation technique. The present study compares the detection limit and sensitivity of a technique based on the use of commercially available magnetic fractionation columns with those for thick blood film microscopy and reverse transcriptase polymerase chain reaction (RT-PCR) methods.

METHODS

Gametocyte detection in six series of dilutions of cultured P. falciparum parasites with known gametocytaemia was conducted using magnetic fractionation, thick blood film, and RT-PCR techniques.

RESULTS

The preparations obtained by the magnetic fractionation method were of thin film quality allowing easy gametocyte identification by light microscopy. Magnetic fractionation had a higher sensitivity and approximately two orders of magnitude better limit of detection than thick blood film microscopy. Gametocytes were also more readily detectable on the magnetically fractionated preparations. Magnetic fractionation had a similar limit of detection to that of RT-PCR.

CONCLUSION

Magnetic fractionation is a highly sensitive and convenient method for gametocyte detection in comparison with the standard thick blood film and RT-PCR methods, and could readily be adapted to field application.

摘要

背景

疟原虫感染红细胞的磁性特性已被用于不同的临床和研究目的。最近在巴布亚新几内亚农村临床环境中的一项研究表明,磁性沉积显微镜有助于检测恶性疟原虫配子体,但尚未有研究确定磁性分离技术的相对灵敏度和检测限。本研究比较了基于使用市售磁性分离柱的技术与厚血膜显微镜检查和逆转录聚合酶链反应(RT-PCR)方法的检测限和灵敏度。

方法

使用磁性分离、厚血膜和RT-PCR技术对已知配子体血症的六组恶性疟原虫培养物稀释液进行配子体检测。

结果

通过磁性分离方法获得的制剂具有薄膜质量,便于通过光学显微镜轻松识别配子体。磁性分离比厚血膜显微镜检查具有更高的灵敏度和大约两个数量级更好的检测限。在磁性分离的制剂上也更容易检测到配子体。磁性分离的检测限与RT-PCR相似。

结论

与标准厚血膜和RT-PCR方法相比,磁性分离是一种用于配子体检测的高度灵敏且方便的方法,并且可以很容易地应用于现场。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c3a4/2689255/6308a81bc593/1475-2875-8-98-1.jpg

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