The Detection Limit of PCR Amplification for Cryptosporidium spp. Oocysts in Fecal Samples

Al-Warid, Harith Saeed; Al-Saqur, Ihsan M.; Mahmood, Souhaila H.

doi:10.1007/s40009-018-0770-7

Cited by 4 publications

(1 citation statement)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Therefore, by relating parasite particle concentration to Ct values, it is normal to observe that the number of cycles decreases when C. parvum load increases, as reported in previous studies. 61,62 We included the current measurement using DPV for the proposed NMI aptasensor with both healthy stool samples (n = 7) and patient stool samples (n = 10) to evaluate its performance in complex media. The patient samples were also randomized with a mixture of both healthy and diseased ones to assess the clinical applicability under realistic settings (Figures 4e,f).…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

Aptamer-Based Electrochemical Microfluidic Biosensor for the Detection of Cryptosporidium parvum

et al. 2023

View full text Add to dashboard Cite

Cryptosporidium parvum is a high-risk and opportunistic waterborne parasitic pathogen with highly infectious oocysts that can survive harsh environmental conditions for long periods. Current state-of-the-art methods are limited to lengthy imaging and antibody-based detection techniques that are slow, labor-intensive, and demand trained personnel. Therefore, the development of new sensing platforms for rapid and accurate identification at the point-of-care (POC) is essential to improve public health. Herein, we propose a novel electrochemical microfluidic aptasensor based on hierarchical 3D gold nano-/ microislands (NMIs), functionalized with aptamers specific to C. parvum. We used aptamers as robust synthetic biorecognition elements with a remarkable ability to bind and discriminate among molecules to develop a highly selective biosensor. Also, the 3D gold NMIs feature a large active surface area that provides high sensitivity and a low limit of detection (LOD), especially when they are combined with aptamers,. The performance of the NMI aptasensor was assessed by testing the biosensor's ability to detect different concentrations of C. parvum oocysts spiked in different sample matrices, i.e., buffer, tap water, and stool, within 40 min detection time. The electrochemical measurements showed an acceptable LOD of 5 oocysts mL −1 in buffer medium, as well as 10 oocysts mL −1 in stool and tap water media, over a wide linear range of 10−100,000 oocysts mL −1 . Moreover, the NMI aptasensor recognized C. parvum oocysts with high selectivity while exhibiting no significant cross-reactivity to other related coccidian parasites. The specific feasibility of the aptasensor was further demonstrated by the detection of the target C. parvum in patient stool samples. Our assay showed coherent results with microscopy and real-time quantitative polymerase chain reaction, achieving high sensitivity and specificity with a significant signal difference (p < 0.001). Therefore, the proposed microfluidic electrochemical biosensor platform could be a stepping stone for the development of rapid and accurate detection of parasites at the POC.

show abstract

Section: ■ Experimental Sectionmentioning

confidence: 99%