CN210665482U - Helicobacter pylori detection device based on CN free radical isotope spectrum - Google Patents
Helicobacter pylori detection device based on CN free radical isotope spectrum Download PDFInfo
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- CN210665482U CN210665482U CN201921671923.4U CN201921671923U CN210665482U CN 210665482 U CN210665482 U CN 210665482U CN 201921671923 U CN201921671923 U CN 201921671923U CN 210665482 U CN210665482 U CN 210665482U
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- 150000003254 radicals Chemical class 0.000 title claims abstract description 72
- 238000001514 detection method Methods 0.000 title claims abstract description 65
- 238000001228 spectrum Methods 0.000 title claims abstract description 43
- 241000590002 Helicobacter pylori Species 0.000 title claims abstract description 23
- 229940037467 helicobacter pylori Drugs 0.000 title claims abstract description 23
- 238000012545 processing Methods 0.000 claims abstract description 16
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- 230000003595 spectral effect Effects 0.000 abstract description 50
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 12
- 229910052799 carbon Inorganic materials 0.000 abstract description 12
- 238000006073 displacement reaction Methods 0.000 abstract description 2
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- 239000004202 carbamide Substances 0.000 description 8
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- KZMAWJRXKGLWGS-UHFFFAOYSA-N 2-chloro-n-[4-(4-methoxyphenyl)-1,3-thiazol-2-yl]-n-(3-methoxypropyl)acetamide Chemical compound S1C(N(C(=O)CCl)CCCOC)=NC(C=2C=CC(OC)=CC=2)=C1 KZMAWJRXKGLWGS-UHFFFAOYSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
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- 108010046334 Urease Proteins 0.000 description 1
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Abstract
The utility model relates to a helicobacter pylori detection device based on CN free radical isotope spectrum, wherein, pulse form is arranged to the detection case by survey person's expired gas, and through the plasma of focused laser beam ablation one-tenth, CO in the expired gas2And N in the environment2Combining to generate CN free radical molecules, collecting CN free radical molecule isotope spectrum data by a multi-channel spectrometer, transmitting the CN free radical molecule isotope spectrum data to a data processing computer for data processing to obtain the frequency displacement and the spectral line intensity of the CN free radical molecule spectral line, and further obtaining the abundance information of the carbon element isotope in the expiration of the detected person. The utility model can avoid the complex sample pretreatment, does not need to detect in the vacuum environment, can shorten the detection time,the detection efficiency is improved, and the cost of the detection device is reduced. In addition, the emission spectral line of CN free radical molecule can not receive atomic spectral line interference, and mass spectrometric detection marks the difficulty to the material spectral line that the mass number is close, therefore the utility model discloses a detection accuracy is higher.
Description
Technical Field
The utility model belongs to isotope on-line measuring field relates to carbon isotope abundance and molecular emission spectrum's measurement, especially relates to a helicobacter pylori detection device based on CN free radical isotope spectrum.
Background
Helicobacter pylori is one of the chronic infectious pathogenic bacteria with the highest infection rate in the world population, and is the only microorganism species known to exist in the human stomach. Research shows that helicobacter pylori infection can cause various diseases such as chronic gastritis, gastric and duodenal ulcers, lymphoid tissue lymphoma and gastric adenocarcinoma related to digestive gastric mucosa. Therefore, it is of great significance to discover early helicobacter pylori infection and effectively kill helicobacter pylori through antibiotics.
There are many methods for detecting helicobacter pylori infection, such as biopsy, isolated culture of helicobacter pylori, rapid urease test, urea breath test, urinary ammonia excretion test, serology test, polymerase chain reaction, etc. At present, the mainstream detection method adopted by hospitals is urea [ 2 ]13C]A method of detecting expiration by orally administering urea to a patient13C]Capsules, if helicobacter pylori is present in the stomach of a patient, this pathogen secretes urease to hydrolyse urea, which after hydrolysis forms carbon dioxide which enters the lungs with the blood and is excreted by respiration, and which can subsequently be detected in the exhaled breath of the patient13Isotope of C. The detection method is safe, accurate, convenient, painless, non-invasive and non-cross-infection, and is known as one of effective methods for detecting helicobacter pylori.
In the above urea [ 2 ]13C]The most critical step in breath test methodsThe carbon isotope (C)13C) And (4) accurately detecting the content. The existing isotope detection means mainly depend on mass spectrometry detection, although the accuracy of the mass spectrometry detection is higher, the pretreatment process of the isotope detection is complicated, the expired gas of a patient needs to be mixed with carrier gas in advance and then enters a mass spectrometry detection box, meanwhile, the mass spectrometry detection needs to be carried out in vacuum, and the requirement on the detection environment is extremely high. Thus, this method has a long analysis time for the abundance of isotopes. In addition, most of the mass spectrometry detection equipment needs to be imported from foreign countries, and the price is high, so that the cost of the mass spectrometry detection-based expiration detection method is high.
Disclosure of Invention
The utility model provides a helicobacter pylori detection device and a method based on CN free radical isotope spectrum, which have high efficiency, low cost and lower requirements on detection environment, and realizes the urea expiration method of helicobacter pylori,13and C, rapid online detection.
The utility model discloses the technical scheme who adopts does:
helicobacter pylori detection device based on CN free radical isotope spectrum includes: detection case, gas conduit, time schedule controller, multichannel spectrum appearance, data processing computer to and set up pulse valve, Nd in the detection case: YAG pulse laser, focusing lens and fiber probe;
the gas guide pipe extends into the detection box, the pulse valve is installed at the end part of the gas guide pipe, and a blowing nozzle is arranged at the end part of the other end, opposite to the gas guide pipe, of the gas guide pipe; and the Nd: a laser beam emitted by a YAG pulse laser is focused to the outlet position of the pulse valve by the focusing lens, and the fiber-optic probe faces to the outlet position of the pulse valve; the time schedule controller is respectively connected with the Nd: YAG pulse laser and multichannel spectrum appearance link to each other, the fiber optic probe with the multichannel spectrum appearance links to each other, the multichannel spectrum appearance with data processing computer links to each other.
Further, the timing controller is respectively connected with the Nd: YAG pulse laser with the multichannel spectrum appearance passes through optical fiber connection, the fiber optic probe with the multichannel spectrum appearance passes through optical fiber connection, the multichannel spectrum appearance with the data processing computer passes through optical fiber connection, it has a plurality of through-holes that supply optic fibre to pass to open on the detection case.
Furthermore, a filter for absorbing moisture in the gas to be detected is arranged on the gas conduit.
The helicobacter pylori detection method based on CN free radical isotope spectrum comprises the following steps:
step one, the mouthpiece accepts the subject to take urea [ 2 ]13C]The gas exhaled after the capsule is discharged into the detection box in a pulse mode at the pulse valve after absorbing moisture through the filter along the gas conduit;
step two, exhausting exhaled gas molecules in the detection box by Nd: a 1064nm laser beam emitted by YAG pulse laser and focused by a focusing lens is ablated into plasma, and CO in the exhaled air2Molecule and N in the detection Box Environment2Combining molecules to generate CN free radical molecules, collecting CN free radical molecule isotope spectral data by a multi-channel spectrometer through an optical fiber probe, and adjusting a time-control device to eliminate spectral data background noise in the process;
thirdly, the CN free radical molecular isotope spectrum data collected by the multi-channel spectrometer are transmitted to a data processing computer for data processing, so that the frequency displacement and the spectral line intensity of the CN free radical molecular spectrum in the isotope spectrum are obtained, and further the abundance information of the carbon element isotope in the expiration of the detected person is obtained;
the frequency shift Deltav formula of the CN free radical molecular spectrum is as follows:
in the formula (1), the first and second groups,
μ is the approximate mass number of the CN radical molecule, i represents13C and other carbon isotopes, omegaeHexix-eAll are spectral constants, v is the number of vibrating quanta, A' represents the energyA level parameter, A' represents a lower energy level parameter;
the spectral line intensity of the CN free radical molecule spectrum is a characteristic peak area obtained by integrating frequency spectral lines corresponding to the CN free radical molecules;
the method specifically comprises the following steps:
step 3-1, firstly, sequentially carrying out normalization and wavelet transformation denoising treatment on CN free radical molecular isotope spectral data, and then extracting B from denoised spectrum2Σ+(v＝0)→X2Σ+(v ═ 1) energy state transitions,12frequency spectrum corresponding to CN free radical molecule and13frequency spectral lines corresponding to CN free radical molecules are obtained by integration respectively12CN and13the characteristic peak area of CN free radical molecule frequency spectral line, namely spectral line intensity;
step 3-2, calculating12CN and13obtaining a characteristic spectral line frequency theoretical value by using the vibration energy level information of CN free radical molecules, wherein E is the vibration energy level energy, h is a Planck constant, and ν is the vibration frequency of the molecules; for CO with known abundance ratio2Performing experimental measurement on an isotope sample, collecting a CN free radical molecular spectrum of the sample, extracting spectral line intensity at a theoretical value of characteristic spectral line frequency, and determining a calibration curve between the CN free radical spectral line intensity and isotope abundance by using a least square method;
step 3-3, substituting the spectral line intensity obtained in the step 3-1 into the calibration curve in the step 3-2 for comparison, inverting the abundance of the isotope, and determining13The content of C isotope.
Further, in step 3-1, CN radical is B2Σ+(v＝0)→X2Σ+(v ═ 1) transitions of energy states,12corresponding spectral line frequency v of CN free radical molecule1＝8.3624×1014Hz，13The spectral line frequency v of the CN radical molecule corresponding to this transition2＝8.3521×1014Hz。
The beneficial effects of the utility model reside in that:
(1) the utility model discloses an adopt spectral detection to replace current mass spectrum technique to carry out quick accurate detection to the carbon isotope, the expired gas can directly be followed gas conduit and detected in getting into the detection case, has avoided loaded down with trivial details sample pretreatment step, can shorten the check-out time to the helicobacter pylori greatly, also improves detection efficiency simultaneously effectively.
(2) The utility model is based on the emission spectrum of CN free radical molecules to realize the pairing13The detection of C isotope abundance is characterized by that in the laser spectrum most of spectral lines are atomic spectral lines, and the difference of linear form and distribution characteristics of molecular spectral line and atomic spectral line is greater, so that the emission spectral line of CN free radical molecule can not be interfered, so that it not only can prevent the difficulty of calibrating spectral lines of substances whose mass numbers are similar in mass spectrum technique, but also can raise the accuracy of mass spectrum technique13The detection precision of the C isotope abundance, namely the detection precision of the content of the helicobacter pylori.
(3) The mass spectrum technology adopted by the existing detection system needs to detect a sample in a vacuum environment, and the CN free radical isotope spectrum technology adopted by the utility model can be directly carried out in an atmospheric environment, thereby greatly reducing the requirements of experimental environment, simplifying the detection process and facilitating the portability of a detection device; meanwhile, the cost of the detection device is reduced, and the commercialization of the product is facilitated.
Drawings
FIG. 1 is a schematic structural diagram of a helicobacter pylori detection device based on CN free radical isotope spectroscopy according to the present invention;
FIG. 2 is a block diagram showing the flow chart of the method for detecting helicobacter pylori based on CN free radical isotope spectroscopy;
FIG. 3 is a CN radical isotope spectrogram (C:)12CN and13CN radical molecules);
reference numerals: 1-detection box, 2-blowing nozzle, 3-filter, 4-gas conduit, 5-pulse valve, 6-Nd: YAG pulse laser, 7-focusing lens, 8-laser beam, 9-optical fiber probe, 10-optical fiber, 11-time schedule controller, 12-multi-channel spectrometer and 13-data processing computer.
Detailed Description
The utility model discloses mainly be through laser and gaseous material effect, produce CN free radical isotope molecule emission spectrum to realize the accurate on-line measuring of carbon isotope with the help of measuring isotope molecule emission spectrum's frequency shift.
The device and method for detecting helicobacter pylori based on CN free radical isotope spectroscopy of the present invention will be further described with reference to the accompanying drawings and specific embodiments.
The helicobacter pylori detection device based on CN free radical isotope spectroscopy shown in figure 1 comprises: detection case 1, gas conduit 4, pulse valve 5, Nd: YAG pulse laser 6, focusing lens 7, fiber probe 9, time sequence controller 11, multi-channel spectrometer 12 and data processing computer 13. Wherein, pulse valve 5, Nd: YAG pulse laser 6, focusing lens 7 and fiber probe 9 are arranged in the detection box 1, and the time schedule controller 11, multi-channel spectrometer 12 and data processing computer 13 are arranged outside the detection box 1.
As shown in FIG. 2, the helicobacter pylori detection method based on CN free radical isotope spectrum, which adopts the detection device, comprises the following steps:
step one, the mouthpiece 2 receives the subject to take urea [ 2 ]13C]The gas exhaled after the capsule, which has absorbed moisture through the filter 3 along the gas conduit 4, is discharged in pulses into the test chamber 1 at the pulse valve 5.
Step two, arrange the call in the detection box 1The gas molecules are substituted by Nd: a 1064nm laser beam 8 from a YAG pulse laser 6 focused by a focusing lens 7 is ablated into plasma (laser ablation), and polyatomic molecular CO in the gas exhaled at the edge portion of the high-temperature plasma2(mainly comprises12C and13C) and N in the environment in the detection box 12The molecules are combined to generate CN free radical molecules and radiate CN free radical molecular spectral lines (the edge temperature of high-temperature plasma is lower than the central temperature of the high-temperature plasma, the CN free radical molecules with too high temperature cannot exist, and the temperature condition for generating the CN free radicals is only provided at the edge of the plasma with lower temperature). The multichannel spectrometer 12 acquires CN free radical molecular isotope spectral data through the optical fiber probe 9, and in the process, the timing controller 11 is adjusted to set a proper delay time to eliminate the spectral data background noise, and the specific delay time is adjusted and determined.
And step three, the CN free radical molecule isotope spectral data collected by the multi-channel spectrometer 12 are transmitted to the data processing computer 13 for data processing, so that the frequency shift and the spectral line intensity of the CN free radical molecule spectrum in the isotope spectral data are obtained, and further the abundance information of the carbon element isotope in the expiration of the detected person is obtained.
The frequency shift Deltav formula of the CN free radical molecular spectrum is as follows:
in the formula (1), the first and second groups,
μ is the approximate mass number of the CN radical molecule, i represents13C and other carbon isotopes, omegaeAnd XeAll are spectral constants, v is the number of vibrating quanta, A 'represents an upper energy level parameter, and A' represents a lower energy level parameter;
and the spectral line intensity of the CN free radical molecule spectrum is a characteristic peak area obtained by integrating frequency spectral lines corresponding to the CN free radical molecules.
Specifically, the third step includes:
step 3-1, firstFirstly, sequentially carrying out normalization and wavelet transformation denoising treatment on CN free radical molecular isotope spectral data, and then extracting B from denoised spectrum2Σ+(v＝0)→X2Σ+(v ═ 1) energy state transitions,12CN radical molecule corresponding frequency spectrum and13frequency spectral lines corresponding to CN free radical molecules are obtained by integration respectively12CN and13the characteristic peak area of the CN free radical molecule frequency spectral line, namely the spectral line intensity.
In this example, CN radical is B2Σ+(v＝0)→X2Σ+(v ═ 1) transitions of energy states,12corresponding spectral line frequency v of CN free radical molecule1＝8.3624×1014Hz，13The spectral line frequency v of the CN radical molecule corresponding to this transition2＝8.3521×1014Hz, as shown in FIG. 3, the spectrum line corresponding to the transition of CN radical molecular spectrum generates frequency shift due to different carbon isotopes, and the carbon isotopes can be distinguished through the frequency shift Deltav.
Step 3-2, calculating by Gaussian 09 software12CN and13the vibration energy level information of CN free radical molecule is first constructed in software12CN free radical molecular structure, then B3PW91/6-31+ G (d) group components are selected to optimize the molecular structure and output the optimized molecular structure information, and the calculation result shows that12The vibration frequency of CN free radical molecule is 2167.96/cm-1. Then, build in software13CN free radical molecular structure calculated by the same radical group13The vibration frequency of CN molecules is 2122.44/cm-1Obtained from E ═ h ν12CN and13the vibration energy level difference between CN is 0.0056eV, wherein E is the vibration energy level energy, h is the Planck constant, and ν is the vibration frequency of the molecule. Will be provided with12The theoretical vibration energy level difference is subtracted from the energy corresponding to the CN free radical molecular spectral line to obtain the product13And energy corresponding to the molecular characteristic spectral line of the CN free radical and a frequency theoretical value thereof. For CO with known abundance ratio2Performing experimental measurement on isotope sample, collecting CN free radical molecular spectrum, and extracting characteristic spectral line frequency theoryThe spectral line intensity at theoretical value is obtained, and a calibration curve between the CN free radical spectral line intensity and the isotope abundance is determined by using a least square method. The calibration curve is established as the relation curve of the abundance ratio and the information of the line intensity, namely once the line intensity is known, the abundance ratio of the carbon isotope can be known.
Step 3-3, substituting the spectral line intensity obtained in the step 3-1 into the calibration curve in the step 3-2 for comparison, and inverting the abundance of the isotope to obtain13The content of C isotope.
The above description is only for the specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto, and any replacement or change method that can be easily conceived by those skilled in the art within the technical scope of the present invention should be covered by the protection scope of the present invention.
Claims (3)
1. Helicobacter pylori detection device based on CN free radical isotope spectrum, characterized by comprising: detection case (1), gas conduit (4), time schedule controller (11), multichannel spectrum appearance (12), data processing computer (13) to and pulse valve (5), Nd of setting in detection case (1): YAG pulse laser (6), focusing lens (7) and fiber probe (9);
the gas guide pipe (4) extends into the detection box (1), the pulse valve (5) is arranged at the end part of the gas guide pipe (4), and the blowing nozzle (2) is arranged at the end part of the other end of the gas guide pipe (4) opposite to the blowing nozzle; nd: a laser beam (8) emitted by a YAG pulse laser (6) is focused to the outlet position of the pulse valve (5) by a focusing lens (7), and an optical fiber probe (9) faces to the outlet position of the pulse valve (5); the timing controller (11) is respectively connected with Nd: YAG pulse laser (6) and multichannel spectrum appearance (12) link to each other, and fiber probe (9) link to each other with multichannel spectrum appearance (12), and multichannel spectrum appearance (12) link to each other with data processing computer (13).
2. The helicobacter pylori detection device based on CN free radical isotope spectroscopy according to claim 1, wherein the timing controller (11) is respectively connected to Nd: YAG pulse laser (6) and multichannel spectrum appearance (12) pass through optic fibre (10) and connect, and optic fibre probe (9) pass through optic fibre (10) with multichannel spectrum appearance (12) and connect, and multichannel spectrum appearance (12) pass through optic fibre (10) with data processing computer (13) and connect, and it has a plurality of through-holes that supply optic fibre (10) to pass to open on detection case (1).
3. The helicobacter pylori detection apparatus based on CN free radical isotope spectroscopy according to claim 1 or 2, wherein the gas conduit (4) is further provided with a filter (3) for absorbing moisture in the gas to be detected.
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| CN110487731B (en) * | 2019-10-08 | 2024-04-19 | 南京信息工程大学 | Helicobacter pylori detection device and method based on CN free radical isotope spectrum |
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