WO2023003524A1 - Screening kit for risk of insecticide exposure - Google Patents
Screening kit for risk of insecticide exposure Download PDFInfo
- Publication number
- WO2023003524A1 WO2023003524A1 PCT/TH2021/000043 TH2021000043W WO2023003524A1 WO 2023003524 A1 WO2023003524 A1 WO 2023003524A1 TH 2021000043 W TH2021000043 W TH 2021000043W WO 2023003524 A1 WO2023003524 A1 WO 2023003524A1
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- WIPO (PCT)
- Prior art keywords
- risk
- screening kit
- insecticide
- insecticide exposure
- exposure according
- Prior art date
Links
- 239000002917 insecticide Substances 0.000 title claims abstract description 58
- 238000012216 screening Methods 0.000 title claims abstract description 26
- 210000004369 blood Anatomy 0.000 claims abstract description 24
- 239000008280 blood Substances 0.000 claims abstract description 24
- 238000006243 chemical reaction Methods 0.000 claims abstract description 17
- 239000000126 substance Substances 0.000 claims abstract description 14
- 239000002904 solvent Substances 0.000 claims abstract description 11
- 238000000926 separation method Methods 0.000 claims abstract description 6
- 238000002360 preparation method Methods 0.000 claims abstract description 4
- 238000012360 testing method Methods 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 102000004190 Enzymes Human genes 0.000 claims description 9
- 108090000790 Enzymes Proteins 0.000 claims description 9
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 238000005259 measurement Methods 0.000 claims description 6
- 210000002966 serum Anatomy 0.000 claims description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 4
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 3
- 239000001110 calcium chloride Substances 0.000 claims description 3
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 3
- 238000001514 detection method Methods 0.000 claims description 3
- 239000000758 substrate Substances 0.000 claims description 3
- NTBLZMAMTZXLBP-UHFFFAOYSA-M 2-acetylsulfanylethyl(trimethyl)azanium;iodide Chemical compound [I-].CC(=O)SCC[N+](C)(C)C NTBLZMAMTZXLBP-UHFFFAOYSA-M 0.000 claims description 2
- GALNBQVDMJRFGJ-UHFFFAOYSA-M 2-butanoyloxyethyl(trimethyl)azanium;iodide Chemical compound [I-].CCCC(=O)OCC[N+](C)(C)C GALNBQVDMJRFGJ-UHFFFAOYSA-M 0.000 claims description 2
- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 2
- 238000005534 hematocrit Methods 0.000 claims description 2
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- 235000011164 potassium chloride Nutrition 0.000 claims description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 2
- XINQFOMFQFGGCQ-UHFFFAOYSA-L (2-dodecoxy-2-oxoethyl)-[6-[(2-dodecoxy-2-oxoethyl)-dimethylazaniumyl]hexyl]-dimethylazanium;dichloride Chemical compound [Cl-].[Cl-].CCCCCCCCCCCCOC(=O)C[N+](C)(C)CCCCCC[N+](C)(C)CC(=O)OCCCCCCCCCCCC XINQFOMFQFGGCQ-UHFFFAOYSA-L 0.000 claims 1
- 238000007664 blowing Methods 0.000 claims 1
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- 238000000034 method Methods 0.000 description 6
- 239000002574 poison Substances 0.000 description 6
- 231100000614 poison Toxicity 0.000 description 6
- 102100033639 Acetylcholinesterase Human genes 0.000 description 4
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- 229940022698 acetylcholinesterase Drugs 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 102000003914 Cholinesterases Human genes 0.000 description 3
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- 229940048961 cholinesterase Drugs 0.000 description 3
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- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 description 2
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- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 208000022535 Infectious Skin disease Diseases 0.000 description 1
- 241000256602 Isoptera Species 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
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- 208000007101 Muscle Cramp Diseases 0.000 description 1
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- 229920001222 biopolymer Polymers 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
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- 230000008859 change Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- YAGKRVSRTSUGEY-UHFFFAOYSA-N ferricyanide Chemical compound [Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] YAGKRVSRTSUGEY-UHFFFAOYSA-N 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
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- 231100000888 hearing loss Toxicity 0.000 description 1
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- 201000001881 impotence Diseases 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 208000021267 infertility disease Diseases 0.000 description 1
- 230000010365 information processing Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
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- 239000003550 marker Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 150000002903 organophosphorus compounds Chemical class 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000000361 pesticidal effect Effects 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
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- 208000017520 skin disease Diseases 0.000 description 1
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- 230000003519 ventilatory effect Effects 0.000 description 1
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- 239000011592 zinc chloride Substances 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
- C12Q1/44—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase
- C12Q1/46—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase involving cholinesterase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2334/00—O-linked chromogens for determinations of hydrolase enzymes, e.g. glycosidases, phosphatases, esterases
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2430/00—Assays, e.g. immunoassays or enzyme assays, involving synthetic organic compounds as analytes
- G01N2430/10—Insecticides
Definitions
- Insecticide exposure is deemed to be an incident that can occur in daily lives, not exclusively to agriculturists, where there may be exposure to insecticides in the quantity exceeding standards. Frequent insecticide use in households or heavy use may, in turn, results in accumulated residuals in the body in the quantity exceeding standards as well.
- Data from the Department of Disease Control revealed survey findings that there were people at risk of illnesses from poisons of insecticides and pesticides, who had a chance of contracting infectious skin diseases as well as a higher risk of chronic diseases in groups of those with low immunity or having blood vessel diseases. It has been found that the morbidity from poisons of insecticidal and pesticidal chemicals is increasing every year.
- risk level assessment of insecticide exposure is another important thing in reducing risk potentials that may lead to physical detriment or cause diseases. Should such risk be detected quickly, lifestyles can be modified, or safe products can be chosen to reduce physical risk.
- Risk level assessment of exposure to insecticides or chemicals varies in its format so that it is right for the operation, right for the specimen and right for the type of substance that needs assessed. Based on exploring, work that has applied for the protection and has similarities to the said work has been found as follows.
- a Thai petty patent application number 1703002151 related to a portable quantitative screening kit for risk of insecticide exposure consists of a reactive unit serving to show a reaction between the blood to be screened and a chemical that is a cholinesterase substrate containing thiocholine, a reaction meter unit with spectrophotometry serving to measure ferricyanide changes of the product from the reactive unit, and an information-processing unit to convert signals from the reaction meter unit so that the chemical reaction changes are visualized.
- the said invention uses an analytical tool with detectable specificity, hence limitation in the measurement.
- a United States patent under the publication number US20110166043A1 is related to a composite method in detecting a marker of exposure of mammals to organophosphate compounds.
- Organophosphate compounds consist of pesticides and metabolites, and include highly reactive organophosphate compounds.
- the biomarker represents the interaction of organophosphate compounds and polypeptides.
- the said invention is different with regard to the process and the reagent used in the detection.
- a United States patent under the publication number US 10913967B2 is related to a system capable of assessing red blood cell AChE function for alerting of exposure and probably suggesting treatment under medical standards, a portable system to oversee in vitro diagnostic points that can detect red blood cell acetylcholinesterase (AChE) from undiluted blood samples as an alert prior to signs of exposure to organophosphorus compound poisons or other poisons.
- the said invention uses different red blood cells as test samples and results in a limitation in that laboratory is required.
- the above-mentioned inventions reveal different assessment processes as per objectives of chemicals that need testing and have a limitation in that laboratory is required for their use, hence the research and development of enzyme-derived assessment kit with accuracy in indicating insecticide exposure levels.
- the developed technique was designed for easy, quick and convenient use with sufficient accuracy in indicating risk levels of insecticide exposure to be used in the daily life of each individual.
- Screening kit for risk of insecticide exposure is a kit that serves to assess the risk of exposure to health-threatening substances with enzyme in assessing insecticide exposure levels in the body. Assessment steps consist of blood sample collection, solvent preparation, sample separation and reaction, and then the solution is tested in a color meter so that exposure risk levels of insecticides accumulated in the body for the past 12-day period can be determined accurately.
- the intent of this invention has been to develop screening kit for risk of insecticide exposure that can assess the exposure risk of health-threatening substances, which may include chemicals, pesticides and insecticides in the field because the said kit has been developed for easy use with accuracy that yields results quickly; and to develop the use at an organization level to facilitate safe work mode as well as contributing to further efficient health check-up welfare.
- Figure 1 shows a part of a sample color meter.
- Figure 2 shows another part of a sample color meter.
- Indication of insecticide exposure risk using enzyme The said enzyme is re-generated every 12 days, and the screening kit for risk of insecticide exposure thereby helps indicate insecticide exposure in the past 12-day period.
- the reaction was designed for easy, quick and convenient use with sufficient accuracy in indicating risk levels of insecticide exposure, suitable for field use and use for the assessment of insecticide exposure risk in households, communities, organizations and people at risk of insecticide exposure, as a tool in monitoring insecticide exposure levels of individuals that can provide assessment results within one to two hours following blood drawing, thereby enabling the use of the risk assessment results in conjunction with risk factor assessment in individuals to assist medical and public health personnel in promoting health or as suggestion in insecticide exposure control and minimize the risk in individuals efficiently.
- the assessment process of the screening kit for risk of insecticide exposure consists of the following steps.
- Solvent preparation Prepare 3 ml solvent at the substance concentration of 0.1-1.5% weight by volume, where the solvent can be chosen from calcium chloride (CaCk), potassium chloride (KC1), dipotassium phosphate (KH2PO4), sodium chloride (NaCl), magnesium chloride (MgCk) or 5,5-dithiobis(2- nitrobenzoic acid).
- the solvent can be chosen from calcium chloride (CaCk), potassium chloride (KC1), dipotassium phosphate (KH2PO4), sodium chloride (NaCl), magnesium chloride (MgCk) or 5,5-dithiobis(2- nitrobenzoic acid).
- substrate being acetylthiocholine iodide at the concentration of 2.50-7.50% weight by volume and butyrylcholine iodide at the concentration of 2.50-7.50% weight by volume and an enzyme cofactor, namely magnesium chloride (MgCk) or zinc chloride (ZnCk), at the concentration of 0.2- 1.5% weight by volume.
- MgCk magnesium chloride
- ZnCk zinc chloride
- Figures 1 and 2 show a sample color meter, where the color meter serves to check colors specific to the screening kit for risk of insecticide exposure.
- the color meter consists of the body 100 provided as a sample tube receptacle with an arranged aperture 101 that supports sample tube insertion, and one side of the body 100 is arranged with a display 102 for showing readings of risk level assessment that can be shown as color bands, numerals, letters or symbols, and one side of the body 100 is arranged with a slot 103 used for connecting with another equipment cable or power supply charging,
- the body 100 is assembled with the base 104 that makes a test tube aperture 105 for receiving a test tube 106 align with or close to the sample tube aperture 101.
- the color measurement consists of at least one bulb 107 as lighting in metering color intensity levels of the test tube 106.
- the assessment of insecticide exposure risk is the color level measurement of solution through measuring only the increase or decrease of the red color and/or the blue color. Subsequently, results from the color meter are shown on the display and interpreted per the instructions in Table 1. This color meter is not limited to measurement in a specified wavelength range and is thereby deemed to have an advantage when compared with a spectrophotometer.
- test result falls in criteria 3-5
- another test should be performed to confirm the result, and if the test result falls in criteria 3-5, it is recommended that the suggestion at the end of Table 1 be followed.
- This test kit is for field use for research or study on changes of insecticide exposure levels/insecticide exposure risk only. 2.
- the enzyme used in this assessment has interfering factors being 1. Hepatic diseases, 2.
- Test result confirmation it is recommended that results be interpreted in conjunction with risk- screening questionnaire, history of underlying diseases, history of insecticide use both in occupation like farm spraying and in households, such as substances to kill ants, termites, flies, weevils and annoying insects.
- Test 1 Two hundred and nine serum samples were selected from a population group with history of insecticide exposure (positive) and a group without history of insecticide exposure (negative). 2. The samples were tested in a spectrophotometer to test the efficiency of the reagent in the screening kit for risk of insecticide exposure in comparison with Ellman reaction, 1 2 which is a standard laboratory technique (spectrophotometer, 405 nm).
- test results demonstrated the efficiency or accuracy at 97.13% while the sensitivity was 98.86%, and the specificity was 95.87%; that is, greater than 95% for all variables tested, leading to a conclusion that they are sufficient to support efficient use of the screening kit for risk of insecticide exposure in classifying insecticide exposure.
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Screening kit for risk of insecticide exposure is the development of kit that serves to assess the risk of exposure to health-threatening substances by assessing insecticide exposure levels in the body. The assessment steps consist of blood sample collection, solvent preparation, sample separation and reaction, and then the solution is tested in a color meter so that exposure risk levels of insecticides accumulated in the body can be determined quickly and accurately.
Description
SCREENING KIT FOR RISK OF INSECTICIDE EXPOSURE
Field of the Invention
Chemical science and agriculture with regard to a screening kit for risk of insecticide exposure
Background of the Invention
Insecticide exposure is deemed to be an incident that can occur in daily lives, not exclusively to agriculturists, where there may be exposure to insecticides in the quantity exceeding standards. Frequent insecticide use in households or heavy use may, in turn, results in accumulated residuals in the body in the quantity exceeding standards as well. Data from the Department of Disease Control revealed survey findings that there were people at risk of illnesses from poisons of insecticides and pesticides, who had a chance of contracting infectious skin diseases as well as a higher risk of chronic diseases in groups of those with low immunity or having blood vessel diseases. It has been found that the morbidity from poisons of insecticidal and pesticidal chemicals is increasing every year. Considering the severity of the said chemicals, it has found that their toxicity can be manifested in two forms, namely the acute one, where signs are present immediately upon chemical exposure, such as nausea, vomiting, headache, myalgia, muscle rigidity and spasm, diarrhea, difficulty breathing, blurred vision or burning eyes; and the chronic one which is caused by chronic chemical exposure and caused by accumulated poisons that lead to diseases or health problems, such as cancer, diabetes mellitus, paresis, paralysis, skin diseases, sterility, congenital deformities, hearing loss, impotence, life-threatening conditions and so on. For example, organophosphates-carbamates have inhibitory activities toward nervous system functions.
As such, risk level assessment of insecticide exposure is another important thing in reducing risk potentials that may lead to physical detriment or cause diseases. Should such risk be detected quickly, lifestyles can be modified, or safe products can be chosen to reduce physical risk. Risk level assessment of exposure to insecticides or chemicals varies in its format so that it is right for the operation, right for the specimen and right for the type of substance that needs assessed. Based on exploring, work that has applied for the protection and has similarities to the said work has been found as follows.
A Thai petty patent application number 1703002151 related to a portable quantitative screening kit for risk of insecticide exposure consists of a reactive unit serving to show a reaction between the blood to be screened and a chemical that is a cholinesterase substrate containing
thiocholine, a reaction meter unit with spectrophotometry serving to measure ferricyanide changes of the product from the reactive unit, and an information-processing unit to convert signals from the reaction meter unit so that the chemical reaction changes are visualized. The said invention uses an analytical tool with detectable specificity, hence limitation in the measurement.
A United States patent under the publication number US20110166043A1 is related to a composite method in detecting a marker of exposure of mammals to organophosphate compounds. Organophosphate compounds consist of pesticides and metabolites, and include highly reactive organophosphate compounds. The biomarker represents the interaction of organophosphate compounds and polypeptides. The biomarker interaction detected by an optical sensor, which consists of a receptor linked to a biopolymer, results in optical reading for detection of change values. The said invention is different with regard to the process and the reagent used in the detection.
A United States patent under the publication number US 10913967B2 is related to a system capable of assessing red blood cell AChE function for alerting of exposure and probably suggesting treatment under medical standards, a portable system to oversee in vitro diagnostic points that can detect red blood cell acetylcholinesterase (AChE) from undiluted blood samples as an alert prior to signs of exposure to organophosphorus compound poisons or other poisons. The said invention uses different red blood cells as test samples and results in a limitation in that laboratory is required.
The above-mentioned inventions reveal different assessment processes as per objectives of chemicals that need testing and have a limitation in that laboratory is required for their use, hence the research and development of enzyme-derived assessment kit with accuracy in indicating insecticide exposure levels. The developed technique was designed for easy, quick and convenient use with sufficient accuracy in indicating risk levels of insecticide exposure to be used in the daily life of each individual.
Summary of the Invention
Screening kit for risk of insecticide exposure is a kit that serves to assess the risk of exposure to health-threatening substances with enzyme in assessing insecticide exposure levels in the body. Assessment steps consist of blood sample collection, solvent preparation, sample separation and reaction, and then the solution is tested in a color meter so that exposure risk levels of insecticides accumulated in the body for the past 12-day period can be determined accurately.
The intent of this invention has been to develop screening kit for risk of insecticide exposure that can assess the exposure risk of health-threatening substances, which may include chemicals, pesticides and insecticides in the field because the said kit has been developed for easy use with accuracy that yields results quickly; and to develop the use at an organization level to facilitate safe work mode as well as contributing to further efficient health check-up welfare.
Brief Description of the Drawings
Figure 1 shows a part of a sample color meter.
Figure 2 shows another part of a sample color meter.
Detailed Description of the Invention
Indication of insecticide exposure risk using enzyme: The said enzyme is re-generated every 12 days, and the screening kit for risk of insecticide exposure thereby helps indicate insecticide exposure in the past 12-day period. The reaction was designed for easy, quick and convenient use with sufficient accuracy in indicating risk levels of insecticide exposure, suitable for field use and use for the assessment of insecticide exposure risk in households, communities, organizations and people at risk of insecticide exposure, as a tool in monitoring insecticide exposure levels of individuals that can provide assessment results within one to two hours following blood drawing, thereby enabling the use of the risk assessment results in conjunction with risk factor assessment in individuals to assist medical and public health personnel in promoting health or as suggestion in insecticide exposure control and minimize the risk in individuals efficiently.
The assessment process of the screening kit for risk of insecticide exposure consists of the following steps.
A. Blood sample collection
- Prick a fingertip for blood with a finger pricking device and fill two blood collection tubes.
- Keep the blood collection tubes with blood collection apparatus to allow the blood to separate into compartments for a duration of 30 minutes.
- Centrifuge the blood collection tubes in a hematocrit centrifuge for a duration of five minutes.
B. Solvent preparation
- Prepare 3 ml solvent at the substance concentration of 0.1-1.5% weight by volume, where the solvent can be chosen from calcium chloride (CaCk), potassium chloride (KC1), dipotassium phosphate (KH2PO4), sodium chloride (NaCl), magnesium chloride (MgCk) or 5,5-dithiobis(2- nitrobenzoic acid).
- Adjust the pH to a range of 7.0-8.50 with sodium hydroxide (NaOH) or hydrochloric (HC1) in reaction tube.
- Add 1 drop of substrate being acetylthiocholine iodide at the concentration of 2.50-7.50% weight by volume and butyrylcholine iodide at the concentration of 2.50-7.50% weight by volume and an enzyme cofactor, namely magnesium chloride (MgCk) or zinc chloride (ZnCk), at the concentration of 0.2- 1.5% weight by volume.
C. Sample separation
- Blow the blood collecting tubes using a dropper to separate the serum.
- Add 1 drop of the serum into the reaction tube.
D. Result examination
- Shake the reaction tube to mix.
- Check with the color meter for the reaction of the solution between the blood sample and the solvent for a duration of 30 seconds to 3 minutes.
Figures 1 and 2 show a sample color meter, where the color meter serves to check colors specific to the screening kit for risk of insecticide exposure. The color meter consists of the body 100 provided as a sample tube receptacle with an arranged aperture 101 that supports sample tube insertion, and one side of the body 100 is arranged with a display 102 for showing readings of risk level assessment that can be shown as color bands, numerals, letters or symbols, and one side of the body 100 is arranged with a slot 103 used for connecting with another equipment cable or power supply charging,
The body 100 is assembled with the base 104 that makes a test tube aperture 105 for receiving a test tube 106 align with or close to the sample tube aperture 101. The color measurement consists of at least one bulb 107 as lighting in metering color intensity levels of the test tube 106. Also, there is a socket 108 used for connecting with another equipment cable or power supply charging, and the socket 108 is arranged with a part that connects with battery 109 as power supply when using the color meter outdoor.
The assessment of insecticide exposure risk is the color level measurement of solution through measuring only the increase or decrease of the red color and/or the blue color. Subsequently, results from the color meter are shown on the display and interpreted per the instructions in Table 1. This color meter is not limited to measurement in a specified wavelength range and is thereby deemed to have an advantage when compared with a spectrophotometer.
Remarks:
1. It is recommended that each test be duplicated for accurate result.
2. In the event of one round of test, if the test result falls in criteria 3-5, another test should be performed to confirm the result, and if the test result falls in criteria 3-5, it is recommended that the suggestion at the end of Table 1 be followed.
Additional Details for Consideration of Use
1. This test kit is for field use for research or study on changes of insecticide exposure levels/insecticide exposure risk only. 2. The enzyme used in this assessment has interfering factors being 1. Hepatic diseases, 2.
Hyperlipidemia, 3. Diabetes mellitus, 4. Hypertension and 5. Renal diseases. Group of population with pathology related to the said factors induce abnormal enzyme levels, and assessment findings may fall in levels 3-5. Therefore, the assessment is not recommended in this population group.
3. Test result confirmation: it is recommended that results be interpreted in conjunction with risk- screening questionnaire, history of underlying diseases, history of insecticide use both in occupation like farm spraying and in households, such as substances to kill ants, termites, flies, weevils and annoying insects.
4. Should subjects have test results that fall in levels 3-5 who are found to have no underlying diseases as per 2. And to have history of insecticide use, recommendations for detoxification and health promotion are as follows.
- Reduce insecticide use in their daily lives and stay away from areas with insecticide use and have insecticide exposure levels re-assessed in 12 days.
- Consider using herbs and health supplements used in detoxification, such as Thunbergia tea and liver health-promoting food, to detoxify existing poisons and promote liver health. Subsequently, have insecticide exposure levels re-assessed every 12 days to monitor changes in insecticide exposure levels in the body.
An efficiency test of the reagent in the screening kit for risk of insecticide exposure consisted of:
Test 1. Two hundred and nine serum samples were selected from a population group with history of insecticide exposure (positive) and a group without history of insecticide exposure (negative).
2. The samples were tested in a spectrophotometer to test the efficiency of the reagent in the screening kit for risk of insecticide exposure in comparison with Ellman reaction,1 2 which is a standard laboratory technique (spectrophotometer, 405 nm).
3. A cut-off value was used that indicated enzyme inhibition based on a study by Rao et al. 2016.3 Test Results
Efficiency Assessment of Reaction Selected for Use in Product Design
Sensitivity or true positive rate:
= true positive/(tme positive + false negative) = (87/(87+l))X100 = 98.86%
Specificity or true negative rate:
= true negative/(tme negative + false positive)
= (116/(116+5)) X 100 = 95.87%
Accuracy (ACC): = (true positive + true negative)/(true positive + false positive + true negative + false negative)
= ((87 + 116)/(87 + 1 + 116 + 5)) X 100 = 97.13%
The test results demonstrated the efficiency or accuracy at 97.13% while the sensitivity was 98.86%, and the specificity was 95.87%; that is, greater than 95% for all variables tested, leading to a conclusion that they are sufficient to support efficient use of the screening kit for risk of insecticide exposure in classifying insecticide exposure.
References
1. Ellman GL, Courtney KD, Andres V, Featherstone RM. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961;7:88-95.
2. Komersova A, Komers K, Cegan A. New Findings about Ellman’ s Method to Determine Cholinesterase Activity. Zeitschrift fur Naturforsch - Sect C J Biosci. 2007;62(1-2):150-154. doi: 10.1515/znc-2007 - 1 -225
3. Rao G, Jyothsna M. Serum Cholinesterase Fevels in Organophosphorous Poisoning Patients on Ventilatory Support. Orig Res Artie Indian J Clin Anaesth. 2016;3(l):52-55. doi: 10.5958/2394- 4994.2016.00012.3
Best Mode of the Invention
As described in Detailed Description of the Invention
Claims
1. A screening kit for risk of insecticide exposure consists of the following steps.
A. Blood sample collection
B. Solvent preparation C. Sample separation
2. The screening kit for risk of insecticide exposure according to claim 1, where the blood sample collection consists of:
- Blood drawing from a fingertip with a finger pricking device and use of fingertip blood collection tubes - Placement of the blood collection tubes with blood collection apparatus pending blood separation into compartments
- Centrifugation of the blood collection tubes in a hematocrit centrifuge
3. The screening kit for risk of insecticide exposure according to claim 1, where a solvent prepared consists of: - Preparation of 3 ml solvent at the substance concentration of 0.1 -1.5% weight by volume
- Adjustment of the pH to a range of 7.0-8.50 with sodium hydroxide (NaOH) or hydrochloric (HC1) in reaction tube
- Addition of 1 drop of substrate with acetylthiocholine iodide at the concentration of 2.50- 7.50 % weight by volume and butyrylcholine iodide at the concentration of 2.50-7.50 % weight by volume and an enzyme cofactor at the concentration of 0.2- 1.5 % weight by volume
4. The screening kit for risk of insecticide exposure according to claim 1, where the sample separation consists of:
- Blowing at the blood collection tubes using a dropper so that the serum is separated
- Addition of 1 drop of the serum into the reaction tube
5. The screening kit for risk of insecticide exposure according to claim 1, where the detection consists of
- The reaction tube shaking to mix
- Measurement with a color meter for the reaction of the solution of the blood sample and the solvent for a duration of 30 seconds to 3 minutes
6. The screening kit for risk of insecticide exposure according to claim 3, where the solvent can be chosen from calcium chloride (CaC12), potassium chloride (KC1), dipotassium phosphate (KH2P04), sodium chloride (NaCl), magnesium chloride (MgC12) or 5,5'-dithiobis(2- nitrobenzoic acid)
7. The screening kit for risk of insecticide exposure is composed of the body (100) provided as a sample tube receptacle with an arranged aperture (101) that supports sample tube insertion. Characteristics are that one side of the body (100) is arranged with a display (102) for showing values of risk level assessment, where the body (100) is assembled with the base (104) that makes a test tube aperture (105) for receiving a test tube (106) align with or close to a sample tube aperture (101), where the color measurement consists of a tube (107).
8. The screening kit for risk of insecticide exposure according to claim 7, where the display (102) shows values of risk level assessment that can be visualized as color bands, numerals, letters or symbols
9. The screening kit for risk of insecticide exposure according to claim 7, where the body (100) is arranged with a slot (103) that may be used for connecting with another equipment cable or power supply charging
10. The screening kit for risk of insecticide exposure according to claim 7, where there is at least one bulb (107) as lighting in metering the color intensity levels of the test tube (106)
11. The screening kit for risk of insecticide exposure according to claim 7, where the body (100) is arranged with a socket (108) used for connecting with another equipment cable or power supply charging 12. The screening kit for risk of insecticide exposure according to claim 11 , where the socket (108) is arranged with a part that connects with battery (109) as power supply when using the color meter outdoor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/TH2021/000043 WO2023003524A1 (en) | 2021-07-21 | 2021-07-21 | Screening kit for risk of insecticide exposure |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/TH2021/000043 WO2023003524A1 (en) | 2021-07-21 | 2021-07-21 | Screening kit for risk of insecticide exposure |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2023003524A1 true WO2023003524A1 (en) | 2023-01-26 |
Family
ID=84978857
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/TH2021/000043 WO2023003524A1 (en) | 2021-07-21 | 2021-07-21 | Screening kit for risk of insecticide exposure |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2023003524A1 (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1116539A (en) * | 1965-01-28 | 1968-06-06 | Horace William Gerarde | Improvements relating to blood testing |
| GB1164255A (en) * | 1967-03-21 | 1969-09-17 | Pfizer & Co C | Improvements in or relating to Diagnostic Package. |
| US20020165620A1 (en) * | 2000-05-05 | 2002-11-07 | Feaster Shawn R. | Assay for detecting, measuring and monitoring the activities and concentrations of proteins and methods of use thereof |
| CN1508262A (en) * | 2002-12-18 | 2004-06-30 | ҽѧ�о�Ժ | Acetylcholinesterase analysis |
| CN101503731A (en) * | 2009-03-19 | 2009-08-12 | 东莞市绿健生物科技有限公司 | Fast detecting reagent kit for toxicity of pesticide residue |
| US20100062455A1 (en) * | 2006-11-27 | 2010-03-11 | Weng Keong Loke | Rapid Detection of Cholinesterase Inhibitors |
| US20130071394A1 (en) * | 2011-09-16 | 2013-03-21 | John K. Troyer | Compositions and combinations of organophosphorus bioscavengers and hyaluronan-degrading enzymes, and methods of use |
-
2021
- 2021-07-21 WO PCT/TH2021/000043 patent/WO2023003524A1/en active Application Filing
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1116539A (en) * | 1965-01-28 | 1968-06-06 | Horace William Gerarde | Improvements relating to blood testing |
| GB1164255A (en) * | 1967-03-21 | 1969-09-17 | Pfizer & Co C | Improvements in or relating to Diagnostic Package. |
| US20020165620A1 (en) * | 2000-05-05 | 2002-11-07 | Feaster Shawn R. | Assay for detecting, measuring and monitoring the activities and concentrations of proteins and methods of use thereof |
| CN1508262A (en) * | 2002-12-18 | 2004-06-30 | ҽѧ�о�Ժ | Acetylcholinesterase analysis |
| US20100062455A1 (en) * | 2006-11-27 | 2010-03-11 | Weng Keong Loke | Rapid Detection of Cholinesterase Inhibitors |
| CN101503731A (en) * | 2009-03-19 | 2009-08-12 | 东莞市绿健生物科技有限公司 | Fast detecting reagent kit for toxicity of pesticide residue |
| US20130071394A1 (en) * | 2011-09-16 | 2013-03-21 | John K. Troyer | Compositions and combinations of organophosphorus bioscavengers and hyaluronan-degrading enzymes, and methods of use |
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