Schistosomiasis electrochemistry sensing quick determination kit, detection method and preparation method of kit

A technology for rapid detection of schistosomiasis, applied in the direction of material electrochemical variables, measuring devices, scientific instruments, etc., can solve the problems of inability to diagnose the disease early, low sensitivity, high missed detection rate, etc., and achieve low production cost, cross-sectional Low reactivity and less reagents

Active Publication Date: 2015-05-20
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to overcome the defects of low sensitivity, time-consuming, laborious, high missed detection rate and inability to carry out early diagnosis of schistosomiasis detection method in the prior art, and to provide a high-sensitivity and specificity detection method. Electrochemical sensing rapid detection kit for schistosomiasis, detection method and preparation method thereof

Method used

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  • Schistosomiasis electrochemistry sensing quick determination kit, detection method and preparation method of kit
  • Schistosomiasis electrochemistry sensing quick determination kit, detection method and preparation method of kit
  • Schistosomiasis electrochemistry sensing quick determination kit, detection method and preparation method of kit

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Embodiment 1

[0025] Example 1 Electrochemical detection of schistosomiasis antibodies using printed carbon electrodes as sensing elements

[0026] Printed carbon electrode surface treatment: The printed carbon electrode is scanned by cyclic voltammetry in PB buffer to activate the electrode, the voltage range is -0.3V~0.6V, the scan rate is 0.5V / s, scan for 10 cycles, and then rinse with ultrapure water , blow dry with nitrogen.

[0027] Sensing interface construction: drop 10ul of an aqueous solution containing 200mM EDC and 50mM NHS on the electrode surface, and react at room temperature for 15min to activate the carboxyl group. Add 10ul of the single SEA antigen and the mixed SjE16 and SEA antigens in different proportions to the surface of the electrode respectively, react at room temperature for 2 hours, rinse the electrode with PBST, and dry it with nitrogen gas. Add 50 ul of 1% BSA dropwise to the surface of the electrode to seal at room temperature for 1 hour, then rinse the elect...

Embodiment 2

[0031] The preparation method of embodiment 2 schistosome electrochemical sensing rapid assay kit

[0032] 1. Preparation of electrochemical sensing array: Rinse the multi-channel printed carbon paste electrode with ultrapure water, add 50ul10mM phosphate buffer (PB) dropwise to cover the three electrodes, and perform cyclic voltammetry on a multi-channel electrochemical detector To scan, rinse the electrodes with ultrapure water. Then drop 10ul carboxyl activation solution containing 200mM carbodiimide (EDC) and 50mM N-hydroxysuccinimide (NHS) on the surface of the working electrode, react at room temperature for 15 minutes, rinse the electrode with ultrapure water, and then On the surface of the working electrode, drop 5ul containing 50mg / L Schistosoma japonicum recombinant antigen SjE16 (provided by the Chinese Center for Disease Control and Prevention of Parasitic Diseases) and 6mg / L of Schistosoma japonicum soluble egg antigen SEA (provided by the Chinese Center for Disea...

Embodiment 3

[0039] The detection method of embodiment 3 schistosome antibody

[0040] 1. Sample dilution:

[0041] Qualitative detection: Dilute the serum to be tested 1:100 with the sample diluent, for example, add 1 μl of serum to 99 μl of diluent, mix thoroughly, and do not need to dilute the negative and positive reference substances.

[0042] Semi-quantitative detection: Dilute the serum to be tested at 1:100 with the sample diluent, and then serially dilute to 3200 times (the dilution factor can be appropriately increased or decreased according to the antibody concentration), and the negative and positive controls do not need to be diluted.

[0043] 2. Sample addition reaction:

[0044] 2.1 Add 10ul of diluted sample serum to each working electrode, and test 2 wells in parallel for each sample. At the same time, set 2 wells for negative, positive and blank controls. Take 10ul of negative and positive control substances and drop them on the surface of the working electrode respectiv...

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Abstract

The invention provides a schistosomiasis electrochemistry sensing quick determination kit, a detection method and a preparation method of the kit. The kit comprises an electrochemistry sensing array, an enzyme conjugate operating fluid, an enzyme substrate, a negative comparison product, a positive comparison product, a sample diluent and a scrubbing solution, wherein the electrochemistry sensing array comprises a printing carbon electrode and a surface-co-assembled schistosome antigen layer and an adsorbed sealing agent layer; a schistosome antigen is the mixture of SjE16 and SEA, an enzyme conjugate working solution is a second-antibody BSA (bovine serum albumin) solution containing a horseradish peroxidase mark, and the enzyme substrate is the mixture of tetramethyl benzidine and hydrogen peroxide; and the negative comparison product is normal blood serum, the positive comparison product is schistosome infection blood serum, and the sample diluent and the scrubbing solution are phosphate buffer solutions containing surface active agents. The kit provided by the invention has the advantages that the preparation is simple, the cost is low, the multi-sample analysis is performed, detection sensitivity, specificity and reproducibility are good, and the schistosome electrochemistry sensing quick determination kit and the detection method are hopefully used for screening and monitoring schistosome patients in an affected area.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to an electrochemical sensing rapid detection kit for schistosomiasis, a detection method and a preparation method thereof. Background technique [0002] Schistosomiasis is one of the most widely distributed zoonotic parasitic diseases in the world, which seriously endangers human health and directly affects the development of the global economy. Although schistosomiasis has been controlled to some extent in my country, it is still an important public health problem. At present, the diagnosis of schistosomiasis is still the key to the prevention and control of schistosomiasis. It is to find and confirm the direct or indirect evidence indicating the existence of schistosomiasis in the body, and provide a basis for the clinical diagnosis of schistosomiasis infection. The traditional detection method of schistosomiasis is pathogenic detection such as feces test, but the sensitivity...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/551G01N27/26G01N27/30
Inventor 樊春海宋世平邓王平冯正胡薇
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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