在细胞外囊DNA中快速地检测突变
Md Mofizur Rahman1,2, Lixue Wang1,3,2, Yundi Chen1
1The Pq Laboratory of BiomeDx/Rx, Department of Biomedical Engineering, Binghamton University, Binghamton, NY 13902, USA.
Extracellular vesicles and circulating nucleic acids
|April 10, 2025
概括
一种新的30分钟的DNA突变检测试验使用Cas12a装载脂质体来更快地诊断癌症. 这种无PCR和无测序的方法有助于针对非小细胞肺癌患者的向治疗跟踪.
科学领域:
- 生物技术是生物技术.
- 分子诊断学 分子诊断
- 在瘤学瘤学.
背景情况:
- 传统的癌症突变检测试验通常需要PCR和测序,这耗时且依赖于专用设备.
- 由于对病毒检测的高需求,COVID-19大流行突显了癌症突变分析的挑战,影响了针对性治疗的患者.
- 需要快速,可访问和设备独立的方法来检测癌症突变.
研究的目的:
- 使用CRISPR-Cas12a技术开发一种快速的,无PCR和测序的DNA突变检测试验.
- 为了能够有效地检测细胞外囊 (EVs) 中致癌突变,以改善癌症诊断和监测.
- 为了创建一个用户友好的测试,兼容基本的实验室设备,如微板阅读器.
主要方法:
- 使用了CRISPR-Cas12a复合体和光灭 (FQ) 探头,这些探头被封装在脂质体内.
- 通过膜融合将Cas12a和FQ探针引入瘤衍生的细胞外囊泡 (EVs).
- 激活的Cas12a,在与目标DNA杂交后,诱导FQ探针的跨裂变,产生可量化的光信号.
主要成果:
- 在30分钟内成功检测到EV DNA中的EGFR L858R突变,从而消除了繁忙的DNA提取和净化.
- 在30名晚期非小细胞肺癌 (NSCLC) 患者和10名健康捐赠者的队列中,获得了86.7%的灵敏度,90%的特异性和87.5%的准确性.
- 证明了大约8 × 10 ^ 5 EVs的检测极限,相当于~10%的突变等位基因频率 (MAF),适用于在没有目标放大的情况下检测晚期肺癌中的突变.
结论:
- 开发的基于Cas12a的测定方法提供了一种快速有效的方法来检测EVs中的DNA突变.
- 这种分析消除了PCR,测序和大量样本准备的需要,使其可用于常规诊断.
- 该技术有望为多重和高通量突变检测的未来进步提供希望,促进在家进行自我诊断和治疗监测.
相关概念视频
Rapidly Varying Flow
40
Rapidly varying flow (RVF) in open channels is characterized by abrupt changes in flow depth over a short distance, with the rate of depth change relative to distance often approaching unity. These flows are inherently complex due to their transient and multi-dimensional nature, making exact analysis difficult. However, approximate solutions using simplified models provide valuable insights into their behavior.Key Features of Rapidly Varying FlowRVF is commonly observed in scenarios involving...
40
RACE - Rapid Amplification of cDNA Ends
6.2K
Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...
6.2K
Increased pulse rate
626
Tachycardia is a condition marked by an abnormally fast or irregular heart rate, surpassing the typical resting rate. In adults, tachycardia is characterized by a pulse rate ranging from 100 to 180 beats per minute. The increased heart rate can result in inadequate blood flow to various body parts, ultimately diminishing the oxygen supply to organs and tissues.
Many factors can elevate the risk of developing tachycardia. These include advanced age, a family history of arrhythmias, and an...
Many factors can elevate the risk of developing tachycardia. These include advanced age, a family history of arrhythmias, and an...
626
Hazard Rate
77
The hazard rate, also known as the hazard function or failure rate, is a statistical measure used to describe the instantaneous rate at which an event occurs, given that the event has not yet happened. From a probabilistic perspective, it represents the likelihood that a subject will experience the event in a very small time interval, conditional on surviving up to the beginning of that interval. In terms of frequency, the hazard rate can be viewed as the ratio of the number of events to the...
77
Decreased pulse rate
517
Bradycardia is a medical condition in which the heart rate is slower than normal. It occurs when the heart's natural pacemaker, the sinus node, generates slower electrical impulses than the standard rhythm. In adults, bradycardia is diagnosed when the pulse rate falls below 60 beats per minute, indicating a deviation from the normal heart rate range.
There are specific risk factors that can elevate the likelihood of developing bradycardia. Advanced age is a significant factor, with...
There are specific risk factors that can elevate the likelihood of developing bradycardia. Advanced age is a significant factor, with...
517
Reflex Activity
1.5K
A reflex activity is an automatic, involuntary response to specific stimuli. It is a part of our survival mechanism, designed to protect us from potential harm. For example, when a bright light suddenly shines into our eyes, we instinctively close them or look away. This is a simple reflex activity orchestrated by the nervous system without conscious thought or effort.
A reflex exam is a diagnostic procedure performed by a healthcare professional to evaluate the functionality of a patient's...
A reflex exam is a diagnostic procedure performed by a healthcare professional to evaluate the functionality of a patient's...
1.5K


