Signal amplifying method of leaky surface acoustic wave-binary peptide nucleic acid biosensor

A leaky surface wave, biosensor technology, applied in biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problems of limited detection sensitivity, small phase change value, long reaction time, etc., and shorten the hybridization reaction time. , improve the mass load, the effect of low cost

Inactive Publication Date: 2009-11-25
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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Problems solved by technology

But at the same time, it was found that when the LSAW-bisPNA biosensor is used for the genetic detection of clinical pathogenic microorganisms, due to the small quality

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  • Signal amplifying method of leaky surface acoustic wave-binary peptide nucleic acid biosensor
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  • Signal amplifying method of leaky surface acoustic wave-binary peptide nucleic acid biosensor

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Embodiment Construction

[0022] In order to make the object, technical solution and advantages of the present invention clearer, preferred embodiments of the present invention are described in detail below.

[0023] 1. Experimental materials

[0024] (1) Fabrication of double-ended pair resonant LSAW sensor: in the same piece of lithium tantalate (LiTaO 3 ) two channels for detection and reference are prepared on the crystal, and each channel is composed of an LSAW resonator. LiTaO 3 The crystal is cut and rotated by 36° in the Y direction, and LSAW is propagated in the X direction, and the hybridization reaction between the peptide binary nucleic acid probe and the target nucleic acid sequence is detected by recording the phase change. Gold film and LiTaO 3 Between the crystals is metallic chromium. The interdigital transducer first coats a layer of metal film on the surface of the substrate by vacuum coating, and then uses photolithography to carve the pattern of interdigital electrodes.

[002...

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Abstract

The invention discloses a signal amplifying method of leaky surface acoustic wave-binary peptide nucleic acid biosensor comprising adding RecA protein-complementary single strand DNA probe in hybridization reaction of a binary peptide nucleic acid probe and nucleic acid specimen to react together; the RecA protein-complementary single strand DNA probe can be acquired by: designing and synthesizing two complementary single strand DNAs according to a target DNA sequence so that at least one single strand DNA sequence is adjacent to the binary peptide nucleic acid probe; then co-incubating the RecA protein and two single strand DNAs under existence of ATPrS so that the RecA protein and two single strand DNAs are combined to acquire; the invention uses biological property that the RecA protein-complementary single strand DNA probe can form multi-strand body with bis-PNA/bi-stran DNA compound to improve quality load on surface of the sensor, which effectively improves detection agility of the sensor. The method is simple in operation with short hybridization reaction time and low cost, which is suitable for genetic rapid detection of micro clinical pathogenic microorganism.

Description

technical field [0001] The invention relates to a signal amplification method of a biosensor, in particular to a signal amplification method of a leaky acoustic surface wave-bibody peptide nucleic acid biosensor. Background technique [0002] With the development of molecular biology techniques, rapid gene detection has become one of the important means for early diagnosis of clinical etiology. Nucleic acid sensors will play an important role in the rapid detection of genes due to their high efficiency, sensitivity, specificity, compact structure, economical and practical characteristics. [0003] Nucleic acid sensors immobilize nucleic acid probes that can specifically recognize target DNA on the surface of a solid support, perform hybridization reactions with nucleic acid samples, and use various types of transducers to convert the results of the hybridization reactions into acoustic, optical, electrical, etc. The physical signal detected, thereby realizing the detection ...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王云霞张立群陈鸣府伟灵
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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