CN107643263B - Biomolecule detection method based on optical comb coherent imaging analysis - Google Patents
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Abstract
A biomolecule detection method based on optical comb coherent imaging analysis is characterized in that according to the fact that molecules with different structures do not have the same absorption spectrum, the wave coherence position of an absorption band and the intensity of the absorption band reflect the characteristics of the molecular structure in turn, and the method can be used for identifying the chemical groups or the composition structures of unknown samples; the absorption intensity of the band corresponds to the content of the molecule, so that the optical comb spectrum can be used for quantitative analysis and purity identification. Particularly, the method has the advantages of high reproducibility and high precision, can be used for the spectrum calibration of tens of millions of known biomolecules, and establishes a spectrum standard spectrum library or a molecule fingerprint spectrum library as the fingerprint spectrum of the biomolecules. The non-contact nondestructive detection can be carried out on the sample; carry out spectral analysis through high accuracy optical comb to the pathogen, can be fast, accurate discernment pathogen's kind and concentration, because optical comb has wide spectral range, can once only detect the pathogen moreover.
Description
Technical Field
The invention belongs to the technical field of biomolecule detection, and particularly relates to a biomolecule detection method based on optical comb coherent imaging analysis.
Background
Most of the golden standards for clinical disease detection require invasive monitoring to obtain diagnosis, for example, diabetes and acute and chronic renal failure require detection of the blood system, detection of renal fibrosis even requires invasive puncture local surgery to be definite, and the acquisition mode of puncture parts and specimens affects diagnosis of conclusions.
Diabetes is a syndrome caused by chronic hyperglycemia, is a relatively common disease caused by insufficient insulin secretion, and firstly causes sugar metabolism disorder due to insufficient insulin, sugar entry into cells is reduced, glycogen synthesis is reduced, sugar hydrolysis is reduced, pentose phosphate pathway is weakened, and cycle of trinuclear nucleic acid is weakened, thereby reducing glucose utilization by tissues such as liver, muscle, fat and the like.
The pathogenesis of chronic renal failure is very complex, recently, it is known as the "uremic toxin theory", which considers that metabolism is the basic guarantee for life maintenance, and the human kidney plays an irreplaceable role in the guarantee, in the case of renal failure, because most of the kidney parenchyma is destroyed, certain toxins which can be excreted through the kidney under normal conditions are accumulated in the body to generate toxic action, and the substances accumulated in the body are uremic toxins.
Acute and chronic renal failure refers to the condition that the human body can not produce urine through the kidney and waste products and excessive water produced by metabolism in the body are discharged out of the body, such as glucose, protein, amino acid, sodium ion, potassium ion, sodium bicarbonate, acid-base balance disorder and the like, and also refers to the disorder of hormones and inflammatory factors such as renin, erythropoietin, active vitamin D3, prostaglandin, IL-6, cystatin C and the like. Uremic kidney tissue is almost completely fibrotic, resulting in loss of kidney function. Clinical early diagnosis and definition of diabetes and kidney diseases are continuously explored, and the blood glucose and serum creatinine absorption peaks in blood samples are found to be sensitive.
Since the concept of double-optical comb spectroscopy is proposed, related scientific research works perform a large number of verification experiments in the near-infrared and intermediate-infrared band research field, infrared absorption spectra are important 'molecular fingerprints' for representing gas molecular characteristics, and compared with a traditional Fourier transform spectrometer containing a Michelson interferometer, the double-optical comb spectrometer has superior performance in the aspects of spectral resolution, sampling time and signal-to-noise ratio, but also has the defect of non-coincidence of wavelengths, and cannot perform qualitative or quantitative analysis on blood components and gas components.
Disclosure of Invention
In order to solve the above problems, the present invention provides a biomolecule detection method based on optical comb coherent imaging analysis, which realizes the analysis of related blood components and gas components by using an optical comb coherent imaging analyzer.
The invention provides a biomolecule detection method based on optical comb coherent imaging analysis, which is used for detecting the existence of biomolecules in a blood detection material and is characterized by comprising the following steps:
step one, an optical comb coherent imaging analyzer is arranged and adjusted, the analyzer comprises an optical comb synchronous controller (1), a primary optical comb (2), a secondary optical comb (3) and a dielectric reflector (4), the system comprises a 1:1 beam splitter (5), a photoelectric detector (6) and a spectrum analyzer (7), wherein an optical comb synchronous controller (1) is respectively connected with a primary optical comb (2) and a secondary optical comb (3), light emitted from the primary optical comb (2) is reflected by a dielectric reflector (4), light emitted from the secondary optical comb (3) is reflected by the 1:1 beam splitter (5), collimated light beams emitted from the primary optical comb (2) and the secondary optical comb (3) are combined through the dielectric reflector (4) and the 1:1 beam splitter (5) and irradiate the photoelectric detector (6), and the photoelectric detector (6) receives and analyzes converted signals for the spectrum analyzer (7);
opening an optical comb coherent imaging analyzer, and collecting a blank-control optical comb coherent spectrum;
placing a blood detection material between the light paths of the main light comb I (2) and the medium reflector (4), starting an optical comb coherent imaging analysis instrument, and collecting an optical comb coherent spectrum when a sample exists;
step four, calculating the difference of the atlas obtained in the step three and the atlas obtained in the step two to obtain an optical comb coherent atlas of the blood examination material;
and step five, comparing the wave coherence position and the intensity of the absorption band of the optical comb coherence spectrum of the blood sample material with the wave coherence position of the absorption band of the standard biomolecule and the intensity of the absorption band of the standard substance in the step four, and judging whether a certain biomolecule exists in the blood sample material according to the comparison result.
The biomolecule detection method based on the optical comb coherent imaging analysis provided by the invention can also have the following characteristics: wherein the biological molecules include glucose, protein, amino acids, sodium ion, potassium ion, sodium bicarbonate, renin, erythropoietin, active vitamin D3, prostaglandin, IL-6, cystatin C, and other hormones, inflammatory factors, creatinine.
The invention provides an optical combThe biomolecule detection method of coherent imaging analysis can also have the following characteristics: wherein, the wave number range of the primary optical comb I (2) and the secondary optical comb II (3) is 6500 and 550cm-1(ii) a Spectral resolution: 0.5cm-1。
The biomolecule detection method based on optical comb coherent imaging analysis provided by the invention can also have the following characteristics that:
and step six, calculating a relative ratio a/A of the intensity a of the absorption band in the optical comb coherent spectrum of the blood test material to the intensity A of the absorption band of the corresponding standard substance, and calculating to obtain the content a/A of the standard substance contained in the blood test material.
The biomolecule detection method based on the optical comb coherent imaging analysis provided by the invention can also have the following characteristics: wherein the blood sample is a blood smear dried by infrared ray.
The biomolecule detection method based on the optical comb coherent imaging analysis provided by the invention can also have the following characteristics: wherein the biomolecule is creatinine.
Action and Effect of the invention
The invention relates to a biomolecule detection method based on optical comb coherent imaging analysis, which can be used for identifying chemical groups or composition structures of unknown samples according to the fact that molecules with different structures do not have the same absorption spectrum, and the wave coherence position of the absorption band and the intensity of the absorption band reflect the characteristics of the molecular structure; the absorption intensity of the band corresponds to the content of the molecule, so that the optical comb spectrum can be used for quantitative analysis and purity identification; particularly, the method has the advantages of high accuracy and high precision, can be used for the spectrum calibration of tens of millions of known biomolecules, and establishes a spectrum standard spectrum library or a molecule fingerprint spectrum library as a fingerprint spectrum of the biomolecules; the optical comb spectrum has no limitation on the sample, so that the non-contact nondestructive testing can be performed on the sample; the high-precision optical comb is used for carrying out spectral analysis on the pathogens, the types and the concentrations of the pathogens can be rapidly and accurately identified, and the optical comb has a wide spectral range, can be used for detecting the pathogens at one time, has the characteristics of accuracy and high efficiency, and can meet the detection requirements of sudden and new-onset diseases.
Drawings
FIG. 1 is a schematic block diagram of an optical comb coherent imaging analyzer used in the present embodiment;
FIG. 2 spectrum of detection without drying;
FIG. 3 is a spectrum of a dried detection;
FIG. 4 illustrates suspected anomalous bands after gas drying; a is suspected to have a difference at 1682; b is suspected to have a difference at 1613; c is suspected to have a difference at 1567; d is suspected of having a difference at 1535;
FIG. 5 is a chart of the infrared transmission spectra of normal persons and five groups of patients; and
figure 6 infrared absorption spectra of five groups of patients relative to normal human serum creatinine.
Detailed Description
In order to make the technical means, the creation features, the achievement purposes and the effects of the invention easy to understand, the following embodiments specifically describe the biomolecule detection method based on the optical comb coherent imaging analysis in conjunction with the accompanying drawings.
Firstly, clinical experiments and analysis and intervention of the blood experiments are carried out among patients with different treatment schemes of hemodialysis and peritoneal dialysis in chronic renal failure, and pathological and optical imaging detection is also carried out on kidney tumor tissues; the respiratory component detection is performed on patients with clinical diabetes and acute and chronic renal failure. Clinical experiments are carried out on the detection of uremia, hypertension, expiration of patients with diabetes and the detection of serum samples.
The specific detection method will be specifically described below by taking a dried blood smear (prepared by coating blood to be detected on a KBr film and drying the coated blood for 5mins by irradiation with an infrared lamp) as an example.
Step one, setting and adjusting an optical comb coherent imaging analyzer, which comprises an optical comb synchronous controller (1), a primary optical comb (2), a secondary optical comb (3) and a dielectric reflector (4) as shown in figure 1, the system comprises a 1:1 beam splitter (5), a photoelectric detector (6) and a spectrum analyzer (7), wherein an optical comb synchronous controller (1) is respectively connected with a primary optical comb (2) and a secondary optical comb (3), light emitted from the primary optical comb (2) is reflected by a dielectric reflector (4), light emitted from the secondary optical comb (3) is reflected by the 1:1 beam splitter (5), collimated light beams emitted from the primary optical comb (2) and the secondary optical comb (3) are combined through the dielectric reflector (4) and the 1:1 beam splitter (5) and irradiate the photoelectric detector (6), and the photoelectric detector (6) receives and analyzes converted signals for the spectrum analyzer (7);
opening an optical comb coherent imaging analyzer, and collecting a blank-control optical comb coherent spectrum;
placing a blood detection material between the light paths of the main light comb I (2) and the medium reflector (4), starting an optical comb coherent imaging analysis instrument, and collecting an optical comb coherent spectrum when a sample exists;
step four, calculating the difference of the atlas obtained in the step three and the atlas obtained in the step two to obtain an optical comb coherent atlas of the blood examination material;
and step five, comparing the wave coherence position and the intensity of the absorption band of the optical comb coherence spectrum of the blood sample material with the wave coherence position of the absorption band of the standard biomolecule and the intensity of the absorption band of the standard substance in the step four, and judging whether a certain biomolecule exists in the blood sample material according to the comparison result.
As a result, it was found that for the patients with clinical acute and chronic renal failure, the acute renal failure patients without dialysis had no significant difference at 1560 and no relevant abnormality (see fig. 2) when the gas was not dried, but had significant difference at 1560 wavelength band after the gas was dried (see fig. 3), and suspected abnormalities were present at the four suspected spectrum abnormality wavelength bands 1535, 1567, 1613, 1682 (see fig. 4).
After sample detection, the creatinine value of a patient is found to be sensitive, and the value size and the absorption peak have positive correlation. When a patient sample combining cerebrovascular accident and hypertensive diabetes is processed and analyzed (figures 5 and 6), obvious absorption can be observed at 1652 wavenumber and 3100-3600 wavenumber by comparing infrared absorption spectrograms of blood of a patient with that of a normal person (by a subtraction method, the infrared spectrograms of the normal person are subtracted from the infrared spectrograms of the patient), wherein 1652 wavenumber represents an absorption peak of carbonyl (═ C0) in the blood sample, 3100-3600 wavenumber is an absorption peak of amino (-NH2), and the measurement result shows that the creatinine content in the patient is obviously higher than that of the normal person, stronger absorption is shown at a specific group absorption peak, and the height of the absorption peak can also be quantitatively marked to indicate the creatinine content in the patient, so that the method provides a good reference for clinical medical diagnosis. After clinical serological sample comparison, 743 serum Ccr 149umol/L, 730 serum Ccr55umol/L, 717 serum Ccr 48umol/L, 723 serum Ccr 38umol/L and 718 serum Ccr 36umol/L have absorption peak heights consistent with clinical serum sample detection.
The present embodiment relates to a biomolecule detection method based on optical comb coherent imaging analysis, which utilizes optical comb coherent imaging to analyze related blood components and gas components, compares the infrared absorption spectrograms of the blood of a patient and a normal person (by using a difference method, subtracting the infrared spectrogram of the normal person from the infrared spectrogram of the patient), significant absorption was observed at both 1652 wavenumbers and 3100-3600 wavenumbers, wherein 1652 wave number represents the absorption peak of carbonyl (═ C0) in the blood sample, 3100-3600 wave number is the absorption peak of amino (-NH2), and the measurement result shows that the creatinine content in the patient is obviously higher than that in the normal person, the strong absorption is shown at a specific group absorption peak, and the creatinine content in a patient can be indicated quantitatively by the height of the absorption peak, so that a good reference is provided for clinical medical diagnosis. After clinical serological sample comparison, it can be seen that 743 serum Ccr 149umol/L, 730 serum Ccr55umol/L, 717 serum Ccr 48umol/L, 723 serum Ccr 38umol/L, 718 serum Ccr 36umol/L, the serum creatinine values are ranked 723>743>718>730>717, the absorption peak height is consistent with clinical serum sample detection, and a spectrum standard spectrum library or a molecular fingerprint spectrum library for clinical chronic renal function serological detection can be established as a 'fingerprint spectrum' of a biomolecule.
Effects and effects of the embodiments
The invention relates to a biomolecule detection method based on optical comb coherent imaging analysis, which can be used for identifying chemical groups or composition structures of unknown samples according to the fact that molecules with different structures do not have the same absorption spectrum, and the wave coherence position of the absorption band and the intensity of the absorption band reflect the characteristics of the molecular structure; the absorption intensity of the band corresponds to the content of the molecule, so that the optical comb spectrum can be used for quantitative analysis and purity identification; particularly, the method has the advantages of high accuracy and high precision, can be used for the spectrum calibration of tens of millions of known biomolecules, and establishes a spectrum standard spectrum library or a molecule fingerprint spectrum library as a fingerprint spectrum of the biomolecules; the optical comb spectrum has no limitation on the sample, so that the non-contact nondestructive testing can be performed on the sample; the high-precision optical comb is used for carrying out spectral analysis on the pathogens, the types and the concentrations of the pathogens can be rapidly and accurately identified, and the optical comb has a wide spectral range, can be used for detecting the pathogens at one time, has the characteristics of accuracy and high efficiency, and can meet the detection requirements of sudden and new-onset diseases.
The invention provides a detection method for analyzing biomolecules by optical comb coherent imaging, which comprises the following steps: 6500 + 550cm-1(ii) a Spectral resolution: 0.5cm-1(ii) a Wave number accuracy: at 3000cm-1The error range is better than 0.1cm-1(ii) a Signal-to-noise ratio: 9300:1 (integration time 5 s). According to the fact that molecules with different structures do not have the same absorption spectrum, the wave coherence position of an absorption band and the intensity of the absorption band reflect the characteristics of the molecular structure in reverse, the method can be used for identifying the chemical group or the composition structure of an unknown sample, establishing a spectrum standard spectrum library or a molecular fingerprint spectrum library as the fingerprint spectrum of biomolecules, and carrying out non-contact nondestructive detection on the sample.
The detection method for analyzing the biological molecules by the optical comb coherent imaging provided by the invention is corresponding to the content of the molecules by the absorption intensity of a spectral band, so that the optical comb spectrum can be used for quantitative analysis and purity identification.
The detection method for analyzing the biomolecules by the optical comb coherent imaging can be used for the spectrum calibration of tens of millions of known biomolecules according to the measurement advantages of high reproducibility and high precision, and a spectrum standard spectrum library or a molecule fingerprint spectrum library is established to be used as a fingerprint spectrum of the biomolecules.
The optical comb coherent imaging analysis biomolecule detection method provided by the invention has the advantages that the optical comb spectrum has no any limitation on the sample, and the non-contact nondestructive detection can be carried out on the sample.
According to the detection method for analyzing the biomolecules by the optical comb coherent imaging, the optical comb spectrum can quickly and accurately identify the types and the concentrations of pathogens through the established spectrum library.
The detection method for analyzing the biomolecules by the optical comb coherent imaging provided by the invention has the characteristics of accuracy and high efficiency, can detect pathogens at one time due to the wide spectral range of the optical comb, and can meet the detection requirements of sudden and new diseases.
The above embodiments are preferred examples of the present invention, and are not intended to limit the scope of the present invention.
Claims (5)
1. A biomolecule detection method based on optical comb coherent imaging analysis is used for detecting the existence of biomolecules in a blood detection material, and is characterized by comprising the following steps:
step one, setting and adjusting an optical comb coherent imaging analyzer, wherein the analyzer comprises an optical comb synchronous controller (1), a main optical comb I (2), a secondary optical comb II (3), a medium reflector (4), a 1:1 beam splitter (5), a photoelectric detector (6) and a spectrum analyzer (7), the optical comb synchronous controller (1) is respectively connected with the first main optical comb (2) and the second auxiliary optical comb (3), light emitted from the first main optical comb (2) is reflected by a medium reflector (4), light emitted from the second auxiliary optical comb (3) is reflected by a 1:1 beam splitter (5), collimated light beams emitted from the first main optical comb (2) and the second auxiliary optical comb (3) are combined through the medium reflector (4) and the 1:1 beam splitter (5) and irradiate the photodetector (6), the photoelectric detector (6) receives and provides the converted signal for the spectrum analyzer (7) to analyze;
opening the optical comb coherent imaging analyzer, and collecting a blank-control optical comb coherent spectrum;
placing a blood detection material between the light paths of the first main optical comb (2) and the medium reflector (4), starting the optical comb coherent imaging analyzer, and collecting an optical comb coherent spectrum when a sample exists;
step four, calculating the difference of the atlas obtained in the step three and the atlas obtained in the step two to obtain an optical comb coherent atlas of the blood examination material;
step five, comparing the wave coherence position and the intensity of the absorption band of the optical comb coherence map of the blood detection material with the wave coherence position of the absorption band of a standard biomolecule and the intensity of the absorption band of a standard substance in the step four, and judging whether a certain biomolecule exists in the blood detection material according to the comparison result;
the optical comb coherent spectrum is an infrared absorption spectrogram,
the blood test material is a blood smear which is dried by infrared.
2. The biomolecule detection method based on optical comb coherent imaging analysis as claimed in claim 1, wherein:
wherein the biomolecule comprises glucose, protein, amino acids, sodium ions, potassium ions, sodium bicarbonate, renin, erythropoietin, active vitamin D3, prostaglandin, IL-6, cystatin C, inflammatory factors, creatinine.
3. The biomolecule detection method based on optical comb coherent imaging analysis as claimed in claim 1, wherein:
wherein the wave number ranges of the main optical comb I (2) and the auxiliary optical comb II (3) are 6500-550cm-1(ii) a Spectral resolution: 0.5cm-1。
4. The method for detecting biomolecules based on optical comb coherent imaging analysis according to claim 1, further comprising:
and sixthly, calculating a relative ratio a/A of the intensity a of the absorption band in the optical comb coherence spectrum of the blood detection material to the intensity A of the absorption band of the corresponding standard substance, and calculating to obtain that the content of the standard substance in the blood detection material is a/A.
5. The biomolecule detection method based on optical comb coherent imaging analysis as claimed in claim 1, wherein:
wherein the biomolecule is creatinine.
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