WO2008123790A1 - Procédé de diagnostic d'une maladie cancéreuse - Google Patents

Procédé de diagnostic d'une maladie cancéreuse Download PDF

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Publication number
WO2008123790A1
WO2008123790A1 PCT/RU2007/000165 RU2007000165W WO2008123790A1 WO 2008123790 A1 WO2008123790 A1 WO 2008123790A1 RU 2007000165 W RU2007000165 W RU 2007000165W WO 2008123790 A1 WO2008123790 A1 WO 2008123790A1
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WIPO (PCT)
Prior art keywords
sample
spectrum
intensity
cancer
serum
Prior art date
Application number
PCT/RU2007/000165
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English (en)
Russian (ru)
Inventor
Viktor Mikhailovich Mushta
Fedor Vitalyevich Donenko
Vyacheslav Gennadyevich Pevgov
Viktor Vladimirovich Krestinin
Viktor Mikhailovich Putilin
Anton Vladimirovich Sumarokov
Original Assignee
Viktor Mikhailovich Mushta
Fedor Vitalyevich Donenko
Vyacheslav Gennadyevich Pevgov
Viktor Vladimirovich Krestinin
Viktor Mikhailovich Putilin
Anton Vladimirovich Sumarokov
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Viktor Mikhailovich Mushta, Fedor Vitalyevich Donenko, Vyacheslav Gennadyevich Pevgov, Viktor Vladimirovich Krestinin, Viktor Mikhailovich Putilin, Anton Vladimirovich Sumarokov filed Critical Viktor Mikhailovich Mushta
Priority to PCT/RU2007/000165 priority Critical patent/WO2008123790A1/fr
Publication of WO2008123790A1 publication Critical patent/WO2008123790A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/49—Blood
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/28—Investigating the spectrum
    • G01J3/443—Emission spectrometry

Definitions

  • the invention relates to medicine, namely to oncology, and can be used for the diagnosis of cancer, in particular, for the diagnosis of an early stage of cancer, for example, during routine medical examinations.
  • METHOD FOR SELECTION OF FACES FOR IDENTIFICATION OF MALIGNANT NOVALOGINES which is used in medicine, namely, during routine dispensary prophylactic examination of people with the aim of prompt detection of cancer patients in the early stages of tumor development.
  • the method includes taking biological samples from examined samples, isolating Escherichia and streptococcus cultures from it, followed by incubation in the presence of L929 tumor cells, preparing a smear and staining, as well as microscopic examination with the calculation of the diagnostic index of whole cells index / ILC /.
  • an ICC 50-60% of patients are selected at risk, and with an ICI of 49% or lower, in the group of patients
  • SUBSTITUTE SHEET (RULE 26) malignant neoplasms.
  • the disadvantage of this method also lies in its complexity and complexity, which affects the accuracy of the indicators.
  • the METHOD FOR DIFFERENTIAL DIAGNOSTICS OF OBLIGATORY FACILITIES OF THE PREDUCT AND MALIGNANT NEW FORMATIONS [3], which is based on the determination in the human plasma of the presence of particles with hydrodynamic radii of 5-30 nm in the ratio of 35-55%, the obligate form of precancer is diagnosed in the subject. When determining 55% and above, these particles are diagnosed with malignant neoplasms. When determining other ratios of the same particles, they conclude that these diseases are absent. The method allows to identify and evaluate changes in the homeostasis system.
  • the disadvantages of the method [3] is the low information content, which is associated with the fact that biological serum macromolecules: lipoproteins, lipids, proteins in the blood serum are unstable and can themselves form complexes during sample preparation and preparation, the use of physiological saline without stabilizing its acidity when taking measurements.
  • biological serum macromolecules lipoproteins, lipids, proteins in the blood serum are unstable and can themselves form complexes during sample preparation and preparation, the use of physiological saline without stabilizing its acidity when taking measurements.
  • the sampling, the time taken to obtain serum or plasma, the room temperature during the study - all this affects the formation and dissociation of the samples and leads to the fact that this method gives many false positive results.
  • the lack of stabilization of the acidity of physiological saline only enhances the instability of the test sample.
  • the technical result, to which the claimed invention is directed, is to provide reliable diagnosis of cancer, including at an early stage of the disease by obtaining a comparative quantitative characteristic of the radiation intensity during laser correlation spectroscopy of native plasma or blood serum under conditions of monitoring the concentration of salts and the acidity of the solution and acquisition of light scattering intensity spectra for three measurements with control of temperature changes sample sizes and control of particle size changes, in addition, the technical result is the availability, simplicity, cost-effectiveness of the method, as well as high information content, accuracy and visibility of the obtained
  • SUBSTITUTE SHEET (RULE 26) results, which would guarantee the reliability and evidence of the presence or absence of cancer, as well as the possible risk of its occurrence.
  • the proposed method for the diagnosis of cancer including laser correlation spectroscopy of native serum or blood plasma with a comparative determination of the intensity of the emission spectra, characterized in that the initial radiation spectrum of a sample of native serum or blood plasma is obtained at 10-39 0 C, with which the increase in the intensity of the emission spectrum of the specified sample in the buffer solution is compared when it is heated to 70-90 ° C, compared with the original, then a sample of native serum or blood plasma is cooled to 20-40 0 C and the intensity indicators of the spectrum obtained by cooling the sample to 20-40 0 C are compared with the initial radiation spectrum, if there are particles with a size of 150-1000 nm in the resulting radiation spectrum, and fractions of scattered radiation from them less than 2% diagnose a possible risk of detecting cancer, in the presence of these particles with a size of 150 - 1000 nm and fractions of scattered radiation from them more than 2% diagnose the presence of cancer.
  • serum or plasma is obtained from a blood sample, which is analyzed by the radiation intensity during laser correlation spectroscopy. Particles with a size of 150-1000 nm can be detected in the test sample. It is necessary to determine the moment of formation of particles found in the sample: before blood sampling or upon receipt of serum or plasma. To do this, initially obtain the emission spectrum of the sample at 10-39 0 C, then the serum or blood plasma sample is heated to 70-90 0 C. Then, the specified sample is cooled to 20-40 0 C and the intensity indicators of the spectrum obtained by cooling the sample to 20 are compared -40 0 C, with the original emission spectrum. If there are particles with a size of 150 - 1000 nm in the resulting emission spectrum, and the fraction of scattered radiation from them is less than 2%, a possible risk of cancer detection is diagnosed, in the presence of these particles with
  • SUBSTITUTE SHEET (RULE 26) with a size of 150 - 1000 nm and a fraction of the scattered radiation from them more than 2% diagnose the presence of cancer.
  • Biological macromolecules are stabilized in a buffer solution pH7.0 - pH7.6, which consists of a solution of sodium chloride salts and salts of phosphoric acid with an ionic strength equal to the ionic strength of physiological saline, while fixing the temperature at which measurements are made, controlling the temperature during measurements , and change with high accuracy, about 0.3 C, the temperature of the sample.
  • a buffer solution pH7.0 - pH7.6 which consists of a solution of sodium chloride salts and salts of phosphoric acid with an ionic strength equal to the ionic strength of physiological saline
  • the complexes of biological macromolecules dissociate and the radiation intensity changes during laser correlation spectroscopy - they get the first comparison spectrum. If the intensity of the obtained spectrum decreases with respect to the initial spectrum, it is concluded that the obtained spectrum characterizes the presence of cancer risk.
  • it is cooled to 20-40 0 C and a second comparison spectrum is obtained.
  • the spectrum obtained during the measurement process is examined to obtain the distribution function of nanoparticles in size from 1 to 2,000 nm.
  • SUBSTITUTE SHEET (RULE 26) or patient’s blood plasma, which makes it possible to measure precisely changes in serum or blood plasma macromolecules and level the features of preliminary sample preparation.
  • the radiation spectrum of the sample is processed, measuring the particle size distribution, cooled to a temperature range of 20-40 0 C after it is preheated to 70-90 0 C and by the presence of particles with a characteristic size of 150 - 1000 nm, depending on the fraction of scattered radiation, conclusion on the predisposition to cancer.
  • the inventive method allows to detect changes in the state of biological macromolecules, while ensuring high accuracy and information content of the measurements. Studies are performed with a minimum volume of serum or plasma - up to 0.2 ml, the preparative preparation of which allows a comparative analysis of the interaction of biological macromolecules with laser radiation with a change in the temperature of the sample.
  • FIG. Figure 1 presents a table of examples of studying the light scattering intensity of samples by laser correlation spectroscopy of native plasma or blood serum.
  • Table 1 shows the diagnostic data obtained by analyzing the comparative characteristics of the intensities of the entire spectrum, where the same sample taken from the patient was used as a comparison control, which made it possible to measure precisely the changes in the serum or plasma macromolecules of the patient and level the features of the preliminary preparation sample.
  • the accuracy of the diagnostic data shown in Table 1 was confirmed by numerous examples of radiation spectra obtained both from patients who turned out to be healthy and from patients whose diagnosis of a cancer detected by this method was confirmed.
  • the inventive method is as follows. Fasting blood is taken from a person’s vein. Then, serum or plasma is obtained from blood by any of the generally accepted methods. To do this, the test tube with blood is kept until a clot forms, and then the resulting
  • SUBSTITUTE SHEET (RULE 26) blood clot from the walls of the tube in a circular motion.
  • the tube is centrifuged at 300-300Og for 10-30 minutes and the supernatant is transferred to a clean tube, which is either frozen or transferred to the device operator for analysis. If a blood sample was collected in a test tube with an anticoagulant, then after freezing it, it is necessary to remove the fibrin clot from the sample by centrifugation or pipette.
  • hypoisotonic cuvette from 10 to 100% of the isotonicity of physiological saline or from 0.09 to 0.9 g of sodium chloride and phosphoronic acid salt per 100 g of water, phosphate aqueous buffer pH 7.0 - pH 7.8 at temperature 10-39 0 C.
  • the total volume of the sample in the cuvette is 1.0 ml.
  • Laser correlation spectroscopy of the prepared sample is carried out. Spectrum accumulation is carried out for 2-10 minutes. Then this sample is heated to 70-90 0 C, and laser correlation spectroscopy of the prepared sample is carried out. Spectrum accumulation is carried out for 2-10 minutes, thereby obtaining the first comparison spectrum.
  • the sample is cooled until the temperature of the sample returns to 20-40 0 C. And again, laser correlation spectroscopy of the prepared sample is carried out. Spectrum accumulation is carried out for 2-10 minutes, thereby obtaining a second comparison spectrum. Comparing the intensity of the signals of the initial and first comparisons, they conclude that there is a risk of cancer, or the presence of cancer or the absence of cancer. So, if the intensity of the first comparison spectrum is lower than the intensity of the initial spectrum, then a conclusion is made about the risk of cancer or cancer, if the intensity of the first comparison spectrum is higher than the intensity of the initial spectrum, then a conclusion is made that there is no cancer.
  • the second comparison spectrum is analyzed for the presence of particles of 150 - 1000 nm and their fraction in the scattered radiation.
  • the second comparison spectrum is used as a quality standard of the procedure, since the second spectrum of the sample after cooling should coincide with the original spectrum.
  • the patient is 41 years old. Diagnosis: colon cancer, stage IY, metastases to regional lymph nodes, single metastasis to the liver.
  • Donor M 34 years old. During a medical examination of the previous blood donation at the donor point, no pathologies or diseases were detected. The study used blood taken from a donor for routine analyzes required for all voluntary donors.
  • SUBSTITUTE SHEET (RULE 26) conclude that there is no oncological disease, since the intensity of the first comparison spectrum is higher than that of the initial one.
  • the claimed method for the diagnosis of cancer allows you to identify changes in the state of biological macromolecules, while ensuring high accuracy and information. Studies are performed with a minimum volume of blood serum - up to 0.2 ml, the preparative preparation of which allows a comparative analysis of the interaction of biological macromolecules with laser radiation when the temperature of the sample is changed.
  • the invention is aimed at, namely, a method for diagnosing cancer is provided, which provides reliable diagnosis of cancer, including at an early stage of the disease by obtaining a comparative quantitative characteristic of radiation intensity during laser correlation spectroscopy native plasma, or serum under conditions of monitoring the concentration of salts and acidity of the solution and removal light scattering intensity spectra for three measurements with the control of changes in the temperature of the samples and control of the change in particle size, in addition, the distinguishing feature of the method is its availability, simplicity, cost-effectiveness of the method, as well as high information content, accuracy and visibility of the results, which guarantees the reliability and evidence of the absence of cancer, as well as the possible risk of its occurrence.
  • the inventive method for the diagnosis of cancer also has another fundamental difference from the known methods of diagnosis, in which the absolute measurement of any parameters of the sample, for example, the percentage of particles of a certain size, while in the present method receive comparative quantitative characteristics of the radiation intensity when laser correlation spectroscopy of native plasma or serum under conditions of monitoring the concentration of salts and the acidity of the solution and
  • the invention relates to medicine, namely to oncology, and can be used for the diagnosis of cancer, in particular, for the diagnosis of an early stage of cancer, for example, during routine medical examination.
  • a method for diagnosing an oncological disease including laser correlation spectroscopy of native serum or blood plasma with a comparative determination of the intensity of the radiation spectra, differs from the known analogues in that for its implementation, the initial radiation spectrum of a sample of native serum or blood plasma is obtained at 10-39 0 C, s which compare the increase in the intensity of the emission spectrum of the specified sample in a buffer solution when it is heated to 70-90 ° C, compared with the original, then the sample is willow serum or blood plasma is cooled to 20-40 0 C and the spectrum intensity indicators obtained when it is cooled to 20-40 0 C are compared with the initial radiation spectrum, if there are particles with a size of 150 - 1000 nm in the resulting radiation spectrum, and the fraction of the scattered radiation from them less than 2% diagnose a possible risk of detecting
  • the proposed method can be applied in conditions of mass medical examinations of the population in centers adapted to the conditions of work with blood serum, which confirms its industrial applicability.
  • the method is quite simple, affordable, economical, as it does not require large time and financial costs. Using this method, you can select a group of people in need of an in-depth instrumental study in order to detect a tumor focus.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
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  • Chemical & Material Sciences (AREA)
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  • Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Ecology (AREA)
  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention concerne le domaine de la médecine. Le procédé de diagnostic de maladies cancéreuses par spectroscopie à corrélation laser du sérum natif ou du plasma sanguin par rapport à la détermination comparative de l'intensité des spectres de rayonnement. On obtient un spectre initial du rayonnement d'un échantillon du sérum natif ou du plasma sanguin à 10-39°C avec lequel on compare l'augmentation d'intensité du spectre de rayonnement de cet échantillon dans une solution tampon lors de son réchauffement à 70-90°C par rapport au stade initial. L'échantillon de plasma natif est ensuite refroidi à 20-40°C, et les valeurs d'intensité de spectre, obtenues lors du refroidissement de l'échantillon à 20-40°C, sont comparées au spectre de rayonnement initial. En cas de présence dans le spectre de rayonnement obtenu de particules ayant la taille de 150-1000 nm et d'une partie de rayonnement diffus d'au moins 2 % on diagnostique un risque possible de détection d'une maladie cancéreuse. En cas de présence desdites particules ayant la taille de 150-1000 nm et d'une partie de rayonnement diffus de plus de 2 % on diagnostique une maladie cancéreuse.
PCT/RU2007/000165 2007-04-06 2007-04-06 Procédé de diagnostic d'une maladie cancéreuse WO2008123790A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3956695A (en) * 1974-07-22 1976-05-11 Stamm Michael E Microwave spectral identification of cells
RU2105306C1 (ru) * 1996-10-01 1998-02-20 Александр Васильевич Аклеев Способ дифференциальной диагностики облигатных форм предрака и злокачественных новообразований
RU2132635C1 (ru) * 1996-09-30 1999-07-10 Алексеев Сергей Григорьевич Способ диагностики онкологических заболеваний и устройство для его осуществления
US20060099569A1 (en) * 2002-09-30 2006-05-11 Keiko Akimoto Method and device for diagnosing oncological diseases

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3956695A (en) * 1974-07-22 1976-05-11 Stamm Michael E Microwave spectral identification of cells
RU2132635C1 (ru) * 1996-09-30 1999-07-10 Алексеев Сергей Григорьевич Способ диагностики онкологических заболеваний и устройство для его осуществления
RU2105306C1 (ru) * 1996-10-01 1998-02-20 Александр Васильевич Аклеев Способ дифференциальной диагностики облигатных форм предрака и злокачественных новообразований
US20060099569A1 (en) * 2002-09-30 2006-05-11 Keiko Akimoto Method and device for diagnosing oncological diseases

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MIGMANOVA K.L.: "Vozmozhnosti lazernoi korrelyatsionnoi spektroskopii syvorotki krovi v diagnostiki opukholei golovy i shei, avtoreferat na soisk uchen step.kand.med.nauk", ST. PETERSBURG, 2002, pages 1 - 23 *

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