Electrokinetic sandwich assay and DNA mediated charge amplification for enhanced sensitivity and specificity

نویسندگان

چکیده

An electrical immuno-sandwich assay utilizing an electrokinetic-based streaming current method for signal transduction is proposed. The records the changes in current, first when a target molecule binds to capture probes immobilized on inner surface of silica micro-capillary, and then detection interact with bound molecules surface. difference signals these two steps constitute response assay, which offers better selectivity linear concentration dependent within range 0.2–100 nM. proof concept demonstrated by detecting different concentrations Immunoglobulin G (IgG) both phosphate buffered saline (PBS) spiked E. coli cell lysate. A superior specificity sandwich compared corresponding direct along limit 90 pM PBS. prospect improving sensitivity was theoretically analysed, indicated that charge contrast between probe plays crucial role determining signal. This aspect experimentally validated modulating zeta potential conjugating negatively charged DNA oligonucleotides. length conjugated varied from 5 30 nucleotides, altering ?9.3 ± 0.8 mV ?20.1 0.9 mV. measurements showed clear consistent enhancement as function lengths. results presented here conclusively demonstrate electric well further improvement. study therefore step forward developing highly selective sensitive electrokinetic assays possible application clinical investigations.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Sensitivity and Specificity of Nucleic Acid Sequence-Based Amplification Method for Diagnosis of Cutaneous Leishmaniasis

Abstract Background and Objective: Culture, microscopic method is a gold standard method for identification of Lishmania parasite. The use of Molecular methods such as RT- PCR compared to microscopic methods has a higher sensitivity and specificity however, it is not widely used due to its expensive equipment and the time requested. The use of nucleic acid sequence based amplification (NASBA) ...

متن کامل

Tentacle probe sandwich assay in porous polymer monolith improves specificity, sensitivity and kinetics

Nucleic acid sandwich assays improve low-density array analysis through the addition of a capture probe and a specific label, increasing specificity and sensitivity. Here, we employ photo-initiated porous polymer monolith (PPM) as a high-surface area substrate for sandwich assay analysis. PPMs are shown to enhance extraction efficiency by 20-fold from 2 microl of sample. We further compare the ...

متن کامل

بررسی کارایی روشLAMP (Loop-mediated isothermal DNA amplification)

مقدمه:سالمونلاها از مهمترین باکتری های بیماری و عامل تیفوئید، باکتریمی، آنتروکولیت و سالمونلوز است که از عفونت های مشترک انسان و حیوانات می باشد. بیماریهای ناشی از این عامل یک معضل بزرگ بهداشتی در جهان به خصوص در کشورهای در حال توسعه از جمله کشور ما می باشد. بنابراین تشخیص سریع، دقیق و به موقع آن برای جلوگیری از اپیدمی و عوارض مخرب این باکتری ضروری به نظر می رسد. برای تشخیص سالمونلا روش های مخت...

متن کامل

Heterogeneous surface charge enhanced micromixing for electrokinetic flows.

Enhancing the species mixing in microfluidic applications is key to reducing analysis time and increasing device portability. The mixing in electroosmotic flow is usually diffusion-dominated. Recent numerical studies have indicated that the introduction of electrically charged surface heterogeneities may augment mixing efficiencies by creating localized regions of flow circulation. In this stud...

متن کامل

Two methods for increased specificity and sensitivity in loop-mediated isothermal amplification.

The technique of loop-mediated isothermal amplification (LAMP) utilizes four (or six) primers targeting six (or eight) regions within a fairly small segment of a genome for amplification, with concentration higher than that used in traditional PCR methods. The high concentrations of primers used leads to an increased likelihood of non-specific amplification induced by primer dimers. In this stu...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

ژورنال

عنوان ژورنال: Biosensors and Bioelectronics

سال: 2021

ISSN: ['0956-5663', '1873-4235']

DOI: https://doi.org/10.1016/j.bios.2020.112917