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A microfluidic chip for whole blood nucleic acid detection

A microfluidic chip and nucleic acid technology, which is applied in the field of detection and nucleic acid extraction, can solve problems such as aerosol pollution, cross contamination, false positives, etc., and achieve the effects of reducing complexity, simplifying chip design, and avoiding cross contamination of samples

Active Publication Date: 2021-03-19
INTEGRATED BIOSYSTEMS CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the complexity of the nucleic acid extraction process and the sensitivity of amplification detection techniques, cross-contamination, especially aerosol contamination, is prone to occur, leading to false positive results

Method used

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  • A microfluidic chip for whole blood nucleic acid detection
  • A microfluidic chip for whole blood nucleic acid detection
  • A microfluidic chip for whole blood nucleic acid detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Such as figure 1 As shown, a microfluidic chip for whole blood nucleic acid detection disclosed in the present invention consists of two parts: an outer functional area and an inner functional area. Wherein, the outer functional area is circular, and its position can be fixed by the limiting groove (9) in the outer functional area, and the outer functional area includes a purification reagent storage unit, a waste liquid unit and an amplification detection unit.

[0039] Purification reagent storage unit sequentially includes pretreatment liquid chamber (1), sample chamber (2), washing liquid chamber I (3), washing liquid chamber II (4), dealcohol liquid chamber (5), eluent chamber (6 ), and each liquid storage chamber contains a vent hole (7) for injecting reagents or ventilating with the outside world.

[0040] The liquid storage chamber of the purification reagent storage unit is sequentially filled with nucleic acid purification reagents, the pretreatment liquid ch...

Embodiment 2

[0068] The chip structure design and manufacturing method are the same as in Embodiment 1.

[0069] The chip is preloaded with reaction reagents required for nucleic acid purification and amplification detection. Specific reagent types and components are as follows:

[0070] 1) The pretreatment solution chamber contains a pretreatment solution with a pH of 6.5, and its components include 5M guanidine hydrochloride, 10mM Tris-HCl, 1mM EDTA, 1% (v / v) Triton-X100, 0.1% (v / v) Tween20;

[0071] 2) The pH of the lysate is 6.5, and the components include 5M guanidine hydrochloride, 10mM Tris-HCl, 1mM EDTA, 1M sodium acetate, 1% (v / v) Triton-X100, 0.1% (v / v) Tween20;

[0072] 3) Washing liquid I chamber contains washing liquid I, the composition is 80% (v / v) isopropanol;

[0073] 4) Washing liquid II chamber contains washing liquid II, the composition is 70% (v / v) ethanol;

[0074] 5) The dealcohol solution chamber contains dealcohol solution with a pH of 6.5, and its components in...

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Abstract

The invention relates to the technical field of nucleic acid extraction and detection, in particular to a multi-chamber and multi-channel microfluidic chip for whole blood nucleic acid extraction, amplification and detection. The chip can realize the automatic extraction, amplification and detection process of whole blood sample nucleic acid. The chip is composed of an outer functional area and an inner functional area. The outer functional area includes a purification reagent storage unit, a waste liquid unit, and an amplification detection unit; the inner functional area includes a nucleic acid extraction unit, and there is a sealing ring between the outer functional area and the inner functional area. , the protruding microchannel on the sealing ring can realize the connection or closure of the outer functional area and the inner functional area. The chip is pre-installed with reagents for nucleic acid purification, amplification and detection, which can be stored at room temperature and operated in a closed manner to avoid cross-contamination of samples and aerosol contamination of amplification products.

Description

technical field [0001] The invention relates to the technical field of nucleic acid extraction and detection, in particular to a multi-chamber and multi-channel microfluidic chip for whole blood nucleic acid extraction, amplification and detection. Background technique [0002] Whole blood refers to the complete blood components that have not been centrifuged or separated, and it is one of the important sample sources for clinical testing and on-site testing. Because the complex components often interfere with nucleic acid purification and detection methods, it is of great significance to develop a rapid and efficient nucleic acid extraction and purification reagent system based on the composition characteristics of whole blood samples. Commonly used nucleic acid extraction and purification methods include phenol-chloroform extraction, spin column method, and magnetic bead method. The extraction process is complex, cumbersome, time-consuming, and may use toxic reagents, whi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01L3/00
CPCB01L3/502707B01L2300/0803
Inventor 蔡亦梅高静范东雨李洁昆张瑜代有来任鲁风
Owner INTEGRATED BIOSYSTEMS CO LTD
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