Quantitative test of nano gold immunochromatography paper based on photoelectric detection

and information processing technology

Chapter One Introduction

1.1 The purpose and significance of this research

Immunoassay is the method of using antigen and antibody to detect trace substances in
specimens. Based on the specificity and sensitivity of antigen and antibody response, the
application scope of immunoassay covers all categories of medical examination. Any substance
can be detected by immunoassay as long as it can obtain the corresponding specific antibody. As
the position of immunoassay in medical inspection is increasingly urgent, new methods and new
skills have been introduced over the years. On the whole, its eagerness to be anxious is in the
areas of active [1-3] testing and lightweight [4-5] testing.

At present, the methods for fully automated immunoassay are: solid-phase enzyme
immunoassay (Enzyme Linked Immunosorbent Assay, ELISA), time discrimination Fluorescence
Immunoassay (Time Resolved Fluorescence Immunoassay, TRFIA), chemiluminescence
immunoassay (Chemi Luminescence Immunoassay, CLIA) and so on. Enzyme-linked immunoassay
(ELISA analyzer) used to be a more commonly used method in routine tests. This method has a
lower cost and can be used for the determination of a large number of specimens, but the
measurement time is long and the sensitivity is low, which has been basically replaced by other
methods. Time fluorescence immunoassay is often disturbed by background fluorescence of
serum components, test tubes, instrument components, and stray light from the excitation light
source, which affects sensitivity. The most representative is DELFIA, an automatic analyzer
developed and produced by Wallac in Finland. The main difference between the TRFIA analyzer
and the ELISA analyzer is the use of a special time-resolved fluorometer for measurement. The
sensitivity is higher than the ELISA analyzer, which can reach 10-17 / mol, but there are many
interference factors. Chemiluminescence (CLIA) [6-9] is an analytical technique that combines a
highly sensitive chemiluminescence measurement technique with a highly specific immune
response to detect antigens or antibodies. The development of CLIA method has become a
mature advanced technology for the detection of ultra-trace active substances. Its main
advantages are [10]: ① High sensitivity, because it does not require an external light source, it
has a higher signal-to-noise ratio than the fluorescence method, and the lowest 100 molecules
(10-21 / mol) can be detected, the sensitivity is 1 to 2 orders of magnitude greater than ELISA,
which is equivalent to or even more than TRFIA; ② The luminescent marker is stable, and the
validity period can reach several months or even years; ③ The detection range is wide, Up to 6
orders of magnitude; ④ High degree of automation, which avoids errors caused by manual
operations and improves the precision of analysis methods. The immunoassay system developed
by Boehringer Mannheim is called ELECSYS and was put into mass production in 1997. In recent
years, CLIA has developed rapidly, and has become one of the fastest developing and
popularizing immunoassay methods. Its technology is in a leading position in fully automated
solid-phase labeling immunoassays.
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1.2 Research status of immunoassay of gold nanoimmunochromatographic test strips

Nano gold immunochromatographic test strips (referred to as colloidal gold chromatography test
strips or gold standard test strips) mainly use the characteristics of high electron density of
colloidal gold particles with a diameter between 1 and 150 nm. When the colloidal gold marker is
in the corresponding When a large amount of ligands are aggregated, red or pink lines are visible
to the naked eye, so they are often used in qualitative rapid immunoassay methods [16-17]. In
1971, colloidal gold was introduced into immunochemistry, and it was rapidly developed as an
immunolabeling technology [18-19]. Especially in the past 20 years, colloidal gold labeling has
become an important immunolabeling technology, and it has received more and more research.
Attention to the field. At present, immunogold staining at the electron microscope level (IGS),
immunogold staining at the light microscope level (IGSS), and immunogold staining at the
macroscopic level are increasingly becoming powerful tools for scientific research and clinical
diagnosis [20].
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Chapter II Quantitative Test Mechanism of Nanogold Immunochromatographic Test Strips

The quantitative testing process of the nano-gold immunochromatographic test strip is actually a
comprehensive modulation process of biochemical, optical, and electrical. The traditional test
usually only considers the biochemical properties of the test strip, so the corresponding
improvement measures are still only at the level of test strip material selection and biochemical
performance analysis, such as the material of absorbent paper, the material of nitrocellulose
membrane, the size of nano-gold particles And the amount of labeled antibody. Therefore, the
current nano gold immunochromatographic test strips are mainly used for qualitative or semi-
quantitative tests, and the progress of quantitative tests is slow.
In order to speed up the research speed of the quantitative test of the test strip, this paper
combines computer technology, sensing technology, artificial intelligence and other technologies
with biochemical reactions, and proposes a new research idea, that is, trying to use the nanogold
immunochromatographic test strip as a first-level Sensors, fiber optic sensors are used as
secondary sensors, using artificial intelligence and modern information processing technology to
describe the spectral peak characteristic curve generated by the nano-gold
immunochromatographic test strip, and in-depth analysis of various factors affecting the spectral
peak curve to determine the quantitative Test plan.
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2.1 Nano gold immunochromatographic test strip

Nano-gold immunochromatographic test strips for clinical diagnosis have developed rapidly in
recent years due to their advantages of intuitiveness, high detection efficiency, simple method,
no pollution, and low cost. It uses the specific laws of antigen and antibody specificity and the
basic principles of colloidal gold color development. When the test solution (blood, serum, urine,
saliva, depending on the test item) is soaked or dripped into the test strip, the test solution
passes through the water The capillary action of the fiber brings the colloidal gold to the test line
where the antibody is adsorbed. If the antigen in the test solution corresponds to this antibody,
specific binding occurs and the test line is red or purple [114]; otherwise, the test line does not
develop color. Using the photoelectric detection system with the optical fiber sensor as the core
to analyze the spectral shape and peak value of the test line, the concentration of the liquid to be
measured which is positively correlated with the color of the test line can be obtained.
Nanogold refers to fine gold particles, the diameter of which is generally 1 ～ 100nm, and the
surface of the gold particles is negatively charged. Due to the electrostatic repulsive force, it
maintains a stable state in water and forms a stable colloid, so it is also called colloid. gold.
Because the surface of colloidal gold particles has more electric charges, it can adsorb and bind
high molecular substances such as proteins. Using this surface adsorption, the protein is
adsorbed on the surface of the gold sol particles to obtain the colloidal gold-labeled protein.
Because the gold particles have the characteristics of high electron density, and these markers
are aggregated on the solid support to a certain density (ie, when the gold particles are 107 /
mm2), they can appear red to purple to the naked eye. In the 1970s, due to the unique
advantages of nano-gold, it was introduced into immunological experiments as a marker, forming
a gold labeling technology that is one of the four major labeling technologies of modern
immunology, and has been widely used.
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Chapter 3 Wavelet analysis to remove the photoelectric noise of the test strip and extract the
edge characteristics of the test line ................................. 36-58
3.1 Denoising method based on wavelet transform ....................................
3.1.1 Principle of Denoising .............................. 37-39
3.1.2 Wavelet decomposition and reconstruction denoising analysis ........................ 39-41
3.1.3 Denoising analysis of wavelet transform threshold shrinkage method ........................ 41-43
3.1.4 Analysis of translation invariant wavelet denoising ................................ 43-45
3.1.5 Wavelet Transform Modulus Maximum Denoising Analysis ........................ 45-48
3.1.6 Comparison of several wavelet denoising methods ............ 48-49
3.2 Extracting test line edge features ... 49-57
3.2.1 Characterization of Signal Singularity .................... 49-51
3.2.2 Selection principle of wavelet function ................... 51
3.2.3 Select feature scale .......................... 51-52
3.2.4 Implementation of several singularity detection techniques ...
3.2.4.1 Singularity detection method based on multi-resolution analysis ........................ 52
3.2.4.2 Binary discrete wavelet transform singularity detection method ... 52-53
3.2.4.3 Singularity detection method of continuous wavelet transform ......... 53-54
3.2.5 Algorithm implementation of singularity detection based on wavelet transform .................
54-55
3.2.6 Wavelet transform to realize the recognition of the position of the test line ......................
55-57
3.3 Summary .............................. 57-58
Chapter 4 Research on biochemical noise of fuzzy cluster recognition test strip ................ 58-65
4.1 Fuzzy cluster analysis .................. 58-60
4.1.1 Fuzzy C-means algorithm ........... 58-60
4.1.2 Judgment coefficient ........................... 60
4.2 Research on the biochemical noise of the fuzzy cluster recognition test strip ............. 60-64
4.2.1 Principles of Recognition ..................... 60-61
4.2.2 Recognition example ......... 61-64
4.3 Summary ........................ 64-65
Chapter 5 Fitting of standard working curve based on wavelet neural network ... 65-75

Chapter VIII Clinical Test of Nanometer Gold Immunochromatographic Test Strip Intelligent
Quantitative Tester

Clinical Trial (Clinical Trial) is a very intense medical research essentials. The clinical trial of
medical stuff refers to the clinical principle to verify whether the theoretical principles,
fundamental layout, performance and other elements of medical stuff can guarantee its
tranquility and usefulness. Clinical trials are an indispensable step in the research of medical stuff.
Because the tools involved are humans, it is necessary to accept the test methods that have
resonated with the following countries in order to ensure that the research is objective, scientific,
and efficient.
The core issue of standardized clinical trials is to take into account not only the particularity and
complexity of human-oriented subjects, but also the scientific nature of experimental research.
According to the requirements of clinical research design, clinical trials should follow the three
principles of randomization, establishment of control, and blindness.
The nano-gold immunochromatographic test strip intelligent quantitative tester developed by
applying the new ideas of the nano-gold immunochromatographic intelligent quantitative test
introduced in this article has obtained the People's Republic of China Medical Device Registration
Certificate. According to the three principles of clinical trials, the instruments were used in the
Human Chorionic Gonadotropin (HCG), Alpha-Fetoprotein (AFP), and Prostate Specific Antigen
(PSA) in Xiamen Traditional Chinese Medicine Hospital, Shanghai Eighth People ’s Hospital, and
Shanghai Seventh People ’s Hospital. A large number of clinical trials have been done [162-164].
The test results prove that the instrument has important theoretical significance and clinical
application value for quantitative testing of nano-gold immunochromatographic test strips.
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to sum up

This paper combined with the research status of quantitative test of nano-gold
immunochromatographic test strips, deeply analyzed the problems of quantitative test of nano-
gold immunochromatographic test strips. Theoretically study the quantitative testing mechanism
of nano-gold immunochromatographic test strips, focusing on the spectral peak characteristic
curve of the test strips, the nano-gold immunochromatography and the photoelectric detection
technology with optical fiber sensing as the core and modern information processing The
combination of technology has proposed a new concept and new idea of intelligent quantitative
testing of nano-gold immunochromatographic test strips, thereby realizing accurate quantitative
testing of nano-gold immunochromatographic test strips, providing a new and effective method
for clinical immunoassay Means of detection. The research based on the combination of
computer technology, sensor technology, artificial intelligence and other technologies and
biochemical reactions has important theoretical significance and application value. The
innovative work of this thesis mainly includes:
(1) Discuss the quantitative testing mechanism of nano-gold immunochromatographic test strips
and the characteristics of spectral peak signals, analyze in depth the various interference factors
that affect quantitative testing, such as biochemical noise, optical noise, electrical noise, etc., and
establish ideal spectral peaks The mathematical expression of the characteristic curve has laid a
theoretical foundation for the development of a nano-gold immunochromatographic test strip
quantitative tester.
(2) Introduce wavelet transform, fuzzy cluster recognition and wavelet neural network technology
into the quantitative test of nano-gold immunochromatographic test strips, complete the edge
feature extraction of the test strips, noise signal filtering and curve fitting, etc. The analysis of the
signal of the nano-gold immunochromatographic test strip by processing technology has shifted
from theoretical research to practical application. This paper proposes a new method for
intelligent quantitative testing of nano-gold immunochromatographic test strips based on
modern information processing technology, which expands the application of modern
information processing technology in the field of biomedicine.
(3) According to Lamber-Beer law and complementary light principle, research the photoelectric
detection system with fiber optic sensor as the core, and apply the adaptive particle swarm
optimization algorithm to the optimized design of the photoelectric detection system to quantify
the nanogold immunochromatographic test strip The research of testing solves the problem of
obtaining the best signal.

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