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一种用于寄生虫乳酸脱氢酶检测的免疫传感器,作为疟疾生物标志物 - 与商业检测试剂盒的比较。

An immunosensor for parasite lactate dehydrogenase detection as a malaria biomarker - Comparison with commercial test kit.

机构信息

Cranfield University, Cranfield, Bedfordshire MK43 0AL, England, UK.

Cranfield University, Cranfield, Bedfordshire MK43 0AL, England, UK.

出版信息

Talanta. 2018 Sep 1;187:321-329. doi: 10.1016/j.talanta.2018.04.086. Epub 2018 Apr 30.

Abstract

This paper describes the development of an affinity sensor for the detection of Plasmodium falciparum parasite lactate dehydrogenase (pLDH) as one of the biomarkers used for malaria detection. The gold sensor was functionalised with anti-pLDH after cleaning the electrode surface to remove impurities (120 °C, 1 h). The sensor was then treated to block unreacted groups on the surface and minimise matrix interference, before applying it in a sandwich assay to detect pLDH in buffer samples using a dose concentration assay. The sensor was optimised to achieve the best detection sensitivity before using it for pLDH detection in serum samples. The developed sensor achieved a limit of detection (LOD) of 1.80 ng mL and 0.70 ng mL for the detection of pLDH in buffer and in serum samples respectively. The sensor sensitivity was enhanced further with the use of AuNP conjugated to the detection anti-pLDH-enzyme, achieving an LOD of 19 pg mL in buffer and 23 pg mL in serum samples. The performance of the sensor was compared to commercially available Plasmodium immunochromatographic (ICT) malaria kits. The developed sensor was able to detect pLDH in the Dd2 culture medium supernatant at 0.002% parasitaemia without the use of AuNP signal enhancement when compared to the OptiMAL-IT ICT kit (detect pLDH) and the BinaxNOW ICT kit (detection of both pLDH and PfHRP 2) samples. Therefore, the sensor developed in this work is highly sensitive and can be used for pLDH detection for on-site diagnosis of malaria. A cheap and simple device as developed in this work is required to tackle malaria detection.

摘要

本文介绍了一种用于检测恶性疟原虫乳酸脱氢酶(pLDH)的亲和传感器的开发,pLDH 是用于疟疾检测的生物标志物之一。在清洁电极表面以去除杂质(120°C,1 小时)后,金传感器用抗-pLDH 官能化。然后,对传感器进行处理以封闭表面上未反应的基团并最大程度减少基质干扰,然后在夹心测定中在缓冲样品中应用它,使用剂量浓度测定法检测 pLDH。优化传感器以获得最佳检测灵敏度,然后再用于检测血清样品中的 pLDH。开发的传感器在缓冲液和血清样品中检测 pLDH 的检测限(LOD)分别为 1.80ng/mL 和 0.70ng/mL。通过使用与检测抗-pLDH 酶缀合的 AuNP,进一步提高了传感器的灵敏度,在缓冲液中达到 LOD 为 19pg/mL,在血清样品中达到 LOD 为 23pg/mL。将传感器的性能与市售的恶性疟原虫免疫层析(ICT)疟疾试剂盒进行比较。与 OptiMAL-IT ICT 试剂盒(检测 pLDH)和 BinaxNOW ICT 试剂盒(检测 pLDH 和 PfHRP 2)相比,当不使用 AuNP 信号增强时,该传感器能够在 0.002%疟原虫血症的 Dd2 培养基上清液中检测到 pLDH。因此,本工作中开发的传感器具有很高的灵敏度,可用于现场诊断疟疾的 pLDH 检测。需要开发这种廉价且简单的设备来解决疟疾检测问题。

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