Construction method of novel coronavirus nucleic acid detection model based on surface enhanced infrared spectroscopy and principal component analysis
A technology of principal component analysis and infrared spectroscopy, which is applied in the field of construction of the new coronavirus nucleic acid detection model, can solve the problems of long detection time, high detection cost, unfavorable long-term use, etc., and achieve the effect of fast test, low cost and high sensitivity
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Embodiment 1
[0040] A calcium fluoride window was used as the raw material, and a 10 nm Au film was evaporated on it as a surface-enhanced infrared substrate (10 nm Au); compared with a 50 nm flat gold film (50 nm Au). 1 μM of thiol-modified DNA probe solution was deposited on 10 nm Au and 50 nm Au, respectively, and the infrared transmission spectrum was measured. The results are as follows: figure 1 shown. from figure 1 It can be seen that 10 nm Au significantly enhances the IR spectral signals of 1 μM thiol-modified DNA probes.
Embodiment 2
[0042] (1) Preparation of materials:
[0043] test group:
[0044] Surface-enhanced infrared spectroscopy substrate with DNA probes on the surface: the preparation process is as follows: the calcium fluoride window substrate is placed in acetone, absolute ethanol and ultrapure water in sequence for 10 min, and then washed with N 2 Gun drying; 10 nm gold film was evaporated on a clean substrate to form randomly distributed gold nanoparticles; S covalent bond, a single-molecule self-assembly layer is formed on the surface of Au, and the nucleotide sequence of the DNA probe is shown in SEQ ID NO: 1 (p-nCoV-N).
[0045]Nucleic acid single strand solution of the new coronavirus sample: The nucleotide sequence of the nucleic acid single strand of the new coronavirus sample is shown in SEQ ID NO: 2 (t-nCoV-N).
[0046] Nucleic acid single strand solution of healthy sample: The nucleotide sequence of the nucleic acid single strand of the healthy sample is shown in SEQ ID NO: 3 (t-no...
Embodiment 3
[0056] The infrared spectral data of t-nCoV-N and p-nCoV-N collected in Example 2 were processed for smoothing, baseline calibration, curve normalization, and quadratic differentiation, and then processed using the PCA Spectroscopy program in Origin to obtain the following results: image 3 results shown. image 3 a is the scatterplot of the PCA analysis of the t-nCoV-N and p-nCoV-N group data, PC1 and PC2 represent the two principal components with the largest overall difference, and the ellipses represent the 95% confidence interval for each group of samples. from image 3 a It can be seen that a distinction can be made between the t-nCoV-N and p-nCoV-N groups. image 3 b From top to bottom, the original infrared spectrum of t-nCoV-N, the second differential spectrum, the loading plot of PC1 and the loading plot of PC2. from image 3 b It can be seen that the quadratic differential spectrum can reveal all the peaks covered by the broad peaks in the original spectrum, whic...
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