Development of an Aptamer-based Lateral Flow Device for Point-of-care Detection of Toxic Chinese Medicine Herbs (ALPTH Project)
试验速览
- 阶段
- 不适用
- 状态
- 招募中
- 入组人数
- 90
- 试验地点
- 2
- 主要终点
- The accuracy of the newly developed lateral flow device in detecting aconitine, benzoylaconine and aconine in human urine samples
研究概览
简要总结
Aconitine and related alkaloids are potent cardiotoxins and neurotoxins found in the Aconitum species used in traditional Chinese medicine (TCM), such as 'Chuanwu', 'Caowu' and 'Fuzi'. They pose significant health risks when used inappropriately without professional supervision. Patients with acute aconite poisoning often present with a combination of cardiovascular, neurological, and gastrointestinal toxicities, which can occur after an overdose, inadequate processing of the aconite roots, erroneous use of tincture preparations, and contamination or substitution of other herbs with aconite roots. Timely diagnosis of aconite poisoning remains challenging due to the long laboratory turnaround time. The aim of this study is to develop an aptamer-based lateral flow device (LFD) for point-of-care detection of aconitine and its metabolites (benzoylaconine and aconine) and evaluate its diagnostic performance.
详细描述
In this study, the investigators will first develop an aptamer-based LFD for point-of-care detection of aconitine, benzoylaconine and aconine in human urine samples. The investigators will perform in vitro selection of binding aptamers for each of the target alkaloids (aconitine, benzoylaconine and aconine) using systematic evolution of ligands by exponential enrichment (SELEX). High throughput sequencing (HTS) and bioinformatics analysis will be performed to characterise the selected aptamers. The selected aptamers will be integrated into ELISA and LFD competition assay. Spiked samples (80 samples, including 20 for each target alkaloids and 20 controls) will be used for calibrating the LFD reader and determining the cutoff points.
The investigators will then conduct a cross-sectional study on stored human urine samples to evaluate the diagnostic performance of the newly developed LFD. The gold standard will be the independent laboratory analysis by the Centre for PanorOmic Sciences (CPOS) at The University of Hong Kong.
Patient urine samples will be stored at 4°C in emergency departments or Chinese medicine clinics. The samples will then be transported to the Laboratory Block of the School of Biomedical Sciences by our research staff within 12 hours of collection. LFD assays will be performed in batch with 5 replicates. 150 µL aliquots of patients' urine sample will be thawed on ice and added to the LFD sample pad. After the flow has completed (approximately 5 minutes), a LFD reader will be used to scan the line intensities for statistical analysis and diagnosis. The operators of the LFD assays will be blinded to the CPOS analytic results.
The primary outcome is the accuracy of the newly developed LFD in detecting aconitine, benzoylaconine and aconine in human urine samples. The secondary outcomes include other diagnostic metrics, such as sensitivity, specificity, positive and negative predictive values, positive and negative likelihood ratios, the area under the receiver operating characteristic curve and their respective 95% confidence intervals (CIs). The hypothsis is that the LFD can achieve a high diagnostic accuracy, with a sum of the lower bounds of sensitivity and specificity 95% CIs exceeding 1.75.
The study will be conducted in full compliance with the Declaration of Helsinki. The risks of participating in this study will be minimal since the recruited participants only need to submit urine samples. This study will also adhere to the safety guidelines of the University of Hong Kong Biological Safety Policy.
研究设计
- 研究类型
- Observational
- 观察模型
- Case Control
- 时间视角
- Cross Sectional
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •(Aconite Poisoned Group):
- •adult patients aged 18 years or above
- •clinical presentations consistent with acute aconite poisoning, including neurological, gastrointestinal, or cardiovascular symptoms
- •exposure to aconite alkaloids within 3 days
排除标准
- •(Aconite Poisoned Group):
- •refusal of informed consent for urine collection
- •alternative explanations for clinical presentations
- •Inclusion Criteria (Aconite Therapeutic Use Group):
- •adult patients aged 18 years or above
- •documented history of using aconite for therapeutic purposes, as prescribed by a qualified practitioner in a Chinese medicine clinic
- •exposure to aconite alkaloids within 3 days
- •Exclusion Criteria (Aconite Therapeutic Use Group):
- •- Refusal of informed consent for urine collection.
- •Inclusion Criteria (Control Group):
- •adults aged 18 years or above
- •no documented history of exposure to aconitine, benzoylaconine, or aconine within 30 days
- •no clinical evidence of aconite toxicities
- •Exclusion Criteria (Control Group):
- •refusal of informed consent for urine collection
- •recent history of poisoning or overdose from any other substances within 30 days
研究组 & 干预措施
Aconite Poisoned Group
Patients with acute aconite poisoning reported to the Hong Kong Poison Control Centre
Aconite Therapeutic Use Group
Patients with therapeutic use of aconite with no clinical toxicities in Chinese medicine clinics
Control Group
Patients with no exposure to aconite
结局指标
主要结局
The accuracy of the newly developed lateral flow device in detecting aconitine, benzoylaconine and aconine in human urine samples
时间窗: 0 hour
The sum of the lower bounds of the 95% confidence intervals of sensitivity and specificity
次要结局
- True positive(0 hour)
- False positive(0 hour)
- True Negative(0 hour)
- False Negative(0 hour)
- Sensitivity(0 hour)
- Specificity(0 hour)
- Positive likelihood ratio(0 hour)
- Negative likelihood ratio(0 hour)
- Positive predictive values(0 hour)
- Negative predictive value(0 hour)
- The area under the receiver operating characteristic curve(0 hour)
研究者
Lam Pui Kin
Clinical Associate Professor of Practice
The University of Hong Kong