Moradi M, Sattarahmady N, Rahi A, Hatam G R, Sorkhabadi S M Rezayat, Heli H
Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.
Nanomedicine and Nanobiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Physics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Talanta. 2016 Dec 1;161:48-53. doi: 10.1016/j.talanta.2016.08.030. Epub 2016 Aug 9.
Detection of leishmaniasis is important in clinical diagnoses. In the present study, identification of Leishmania parasites was performed by a label-free, PCR-free and signal-on ultrasensitive electrochemical DNA biosensor. Gold nanoleaves were firstly electrodeposited by an electrodeposition method using spermidine as a shape directing agent. The biosensor was fabricated by immobilization of a Leishmania major specific DNA probe onto gold nanoleaves, and methylene blue was employed as a marker. Hybridization of the complementary single stranded DNA sequence with the biosensor under the selected conditions was then investigated. The biosensor could detect a synthetic DNA target in a range of 1.0×10 to 1.0×10molL with a limit of detection of 1.8×10molL, and genomic DNA in a range of 0.5-20ngμL with a limit of detection of 0.07ngμL. The biosensor could distinguish Leishmania major from a non-complementary-sequence oligonucleotide and the tropica species with a high selectivity. The biosensor was applicable to detect Leishmania major in patient samples.
利什曼病的检测在临床诊断中很重要。在本研究中，利什曼原虫的鉴定是通过一种无标记、无PCR且信号开启的超灵敏电化学DNA生物传感器进行的。首先使用亚精胺作为形状导向剂，通过电沉积法电沉积金纳米片。通过将利什曼原虫主要特异性DNA探针固定在金纳米片上来制备生物传感器，并使用亚甲基蓝作为标记物。然后研究了在选定条件下互补单链DNA序列与生物传感器的杂交情况。该生物传感器能够检测浓度范围为1.0×10至1.0×10mol/L的合成DNA靶标，检测限为1.8×10mol/L，以及浓度范围为0.5 - 20ng/μL的基因组DNA，检测限为0.07ng/μL。该生物传感器能够以高选择性区分利什曼原虫主要种与非互补序列寡核苷酸以及热带种。该生物传感器适用于检测患者样本中的利什曼原虫主要种。
