A sample-in- result -out microfluidic system for β-thalassemia diagnostics via direct whole blood PCR-reverse dot blot

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta Pub Date : 2025-05-08 DOI:10.1016/j.cca.2025.120348

Chao Ye , Yan Wei , Yilian Zhao , Tan Tan , Youqiong Li , Xigui Long , Huiyuan Gao , Xiaoxing Zhou , Mengru Xie , Jilin Qing , Zhizhong Chen

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引用次数: 0

Abstract

Objective

Existing thalassemia detection methods demand high − end labs and have complex procedures, leading to long testing cycles. This study aims to develop a convenient detection method integrated with a microfluidic platform for a sample − to − result process. Method: First, optimal conditions for the whole blood direct PCR − reverse dot hybridization (dPCR − RDB) system were explored. Then, its performance was evaluated with clinical samples. Finally, the entire process was integrated into a palm − sized microfluidic chip for “sample − in, result − out” detection. Result: A stable dPCR − RDB system was established. Clinical verification on 149 samples showed a 0.1 μl whole − blood minimum detection limit, 100 % specificity, and resistance to high triglyceride and bilirubin levels. It had 100 % positive and negative coincidence rates with traditional methods (kappa = 1). The microfluidic − integrated platform achieved “sample − in, result − out” with 0.5–1 μl blood in 130 min, sans a PCR lab.

Conclusion

A “sample − in, result − out” microfluidic gene detection platform using whole blood as the template was successfully established.

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一种用于β-地中海贫血直接全血pcr -反向点印迹诊断的进样结果微流控系统

目的现有的地中海贫血检测方法需要高端的实验室和复杂的程序，导致检测周期长。本研究旨在开发一种方便的检测方法集成微流体平台的样品-结果过程。方法：首先，探索全血直接PCR -反向点杂交（dPCR - RDB）系统的最佳条件。然后用临床样品对其性能进行评价。最后，整个过程被集成到一个手掌大小的微流控芯片中，用于“样品输入，结果输出”检测。结果：建立了稳定的dPCR - RDB体系。对149份样品的临床验证显示，该方法的最低检出限为0.1 μl，特异性为100%，对高甘油三酯和胆红素水平具有抗性。与传统方法的正、负符合率均为100% （kappa = 1）。在没有PCR实验室的情况下，微流控集成平台在130分钟内以0.5-1 μl的血液实现了“样品输入，结果输出”。结论成功建立了以全血为模板的“样品进、结果出”的微流控基因检测平台。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinica Chimica Acta 医学-医学实验技术

CiteScore

10.10

自引率

2.00%

发文量

1268

审稿时长

23 days

期刊介绍： The Official Journal of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) Clinica Chimica Acta is a high-quality journal which publishes original Research Communications in the field of clinical chemistry and laboratory medicine, defined as the diagnostic application of chemistry, biochemistry, immunochemistry, biochemical aspects of hematology, toxicology, and molecular biology to the study of human disease in body fluids and cells. The objective of the journal is to publish novel information leading to a better understanding of biological mechanisms of human diseases, their prevention, diagnosis, and patient management. Reports of an applied clinical character are also welcome. Papers concerned with normal metabolic processes or with constituents of normal cells or body fluids, such as reports of experimental or clinical studies in animals, are only considered when they are clearly and directly relevant to human disease. Evaluation of commercial products have a low priority for publication, unless they are novel or represent a technological breakthrough. Studies dealing with effects of drugs and natural products and studies dealing with the redox status in various diseases are not within the journal''s scope. Development and evaluation of novel analytical methodologies where applicable to diagnostic clinical chemistry and laboratory medicine, including point-of-care testing, and topics on laboratory management and informatics will also be considered. Studies focused on emerging diagnostic technologies and (big) data analysis procedures including digitalization, mobile Health, and artificial Intelligence applied to Laboratory Medicine are also of interest.