Determination method

Abstract

To provide a technique for determining a diet action actually performed by a subject.SOLUTION: The determination method of the present invention includes a determination step of determining a diet behavior selected from an restriction for energy intake behavior and an energy consumption promotion behavior performed by a subject within a determination target period, on the basis of an expression level of at least one of genes selected from the group consisting of intake restriction variation genes and consumption promotion variation genes or an expression product thereof, the genes or the expression product being derived from a biological sample of the subject. The intake restriction variation genes are at least one or more genes whose expression level varies depending on restriction for energy intake behavior and whose expression level does not vary depending on energy consumption promoting behavior. The consumption promotion variation genes are at least one or more genes whose expression level does not vary depending on restriction for energy intake behavior but varies depending on energy consumption promotion behavior.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for determining the actual dieting behavior of a subject. [Background technology]

[0002] Attention has been focused on behaviors that are highly effective for dieting and their effects for the purposes of health promotion, obesity relief, beauty, etc. For example, Patent Document 1 discloses a combination containing multiple polynucleotides that are differentially expressed in animals that exhibit a lean phenotype as a result of one or more types of lean phenotype-promoting treatments, including administration of conjugated linoleic acid (CLA), consumption of a high-protein diet, and increased exercise. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Special Publication No. 2012-501175 Summary of the Invention [Problem to be solved by the invention]

[0004] On the other hand, there is no known technology for assessing the dietary behavior (dietary management, increased exercise, etc.) actually undertaken by a subject.

[0005] The present invention relates to a technique for determining diet behavior actually performed by a subject. [Means for solving the problem]

[0006] A determination method according to one aspect of the present invention includes: The method includes a determination step of determining whether the diet behavior is selected from energy intake restriction behavior that restricts energy intake through food and energy consumption promotion behavior that promotes energy consumption by increasing the amount of exercise, which has been performed by the subject within the period to be determined up to the time of collection of the biological sample, based on the expression level of at least one gene selected from the group consisting of intake restriction variable genes and consumption promotion variable genes or their expression products, which is derived from the biological sample of the subject whose diet behavior is to be determined. The intake restriction fluctuating gene is at least one gene whose expression level fluctuates due to the energy intake restriction behavior and whose expression level does not fluctuate due to the energy consumption promoting behavior. The consumption-promoting variable gene is at least one gene whose expression level does not vary due to the energy-intake-restricting behavior, and whose expression level varies due to the energy-consumption-promoting behavior. [Effects of the Invention]

[0007] According to the present invention, it is possible to determine the diet behavior that a subject has actually performed. DETAILED DESCRIPTION OF THE INVENTION

[0008] The present invention is described in detail below. All patents, non-patent documents, and other publications cited herein are hereby incorporated by reference in their entirety.

[0009] [Summary of the Invention] The present invention is characterized by assessing the diet behavior undertaken by a subject based on the expression level of a predetermined gene or its expression product derived from the subject's biological sample. The present invention arose from the discovery of genes whose expression changes specifically before and after dietary and exercise interventions in a subject group, and was derived from the novel idea of ​​assessing the diet behavior undertaken by a subject using the expression levels of these genes or expression products. One of the advantages of the present invention is that the diet behavior undertaken (energy intake restriction behavior and / or energy consumption promotion behavior) can be objectively assessed from the genetic information, which is the biological information of each subject, thereby providing useful information for providing weight loss guidance tailored to each subject.

[0010] In one embodiment of the present invention, the term "gene" refers to double-stranded DNA including human genomic DNA, as well as single-stranded DNA (positive strand) including cDNA, single-stranded DNA (complementary strand) having a sequence complementary to the positive strand, and fragments thereof, and refers to DNA containing some biological information in the sequence information of the bases that make up the DNA. Furthermore, the term "gene" does not only refer to "genes" represented by a specific base sequence, but also includes nucleic acids encoding their homologs (i.e., homologs or orthologs), mutants such as genetic polymorphisms, and derivatives.

[0011] In one embodiment of the present invention, the term "expression product" of a gene encompasses both transcription products and translation products of the gene. A "transcription product" refers to RNA produced by transcription from a gene (DNA), and a "translation product" refers to a protein encoded by the gene that is translated and synthesized based on the RNA. "RNA" includes total RNA, mRNA, rRNA, tRNA, non-coding RNA, and synthetic RNA. In one embodiment of the present invention, the expression level of a target molecule may be measured using RNA, DNA encoding the RNA, a protein encoded by the RNA, a molecule that interacts with the protein, a molecule that interacts with the RNA, or a molecule that interacts with the DNA. RNA is preferred, and mRNA is more preferred. Examples of molecules that interact with RNA, DNA, or proteins include DNA, RNA, proteins, polysaccharides, oligosaccharides, monosaccharides, lipids, fatty acids, and their phosphorylations, alkylations, and sugar adducts, as well as complexes of any of the above.

[0012] In one embodiment of the present invention, the expression level of a gene or its expression product comprehensively means the expression amount or activity level of the gene or expression product, and is indicated, for example, by an index showing the expression amount, an index showing the expression activity, or a level representing these indexes in a graded manner. The index showing the expression level can be appropriately selected depending on the target for measuring the expression level, the measurement method, etc.

[0013] In one embodiment of the present invention, the expression level of a gene or its expression product is derived from a biological sample from a subject. The phrase "the expression level of a gene or its expression product is derived from a biological sample" refers to the expression level being measured or detected from the biological sample. Furthermore, a "biological sample from a subject" refers to a biological sample collected from a subject. The biological sample may be, for example, cells, body fluids (e.g., blood), urine, secretions (e.g., saliva, skin surface lipids), etc., but non-invasively collectable samples (e.g., urine, secretions), etc., are preferred, and skin surface lipids are particularly preferred. Here, "skin surface lipids (SSL)" refers to the fat-soluble fraction present on the surface of the skin and is sometimes called sebum. Generally, SSL mainly contains secretions from exocrine glands such as sebaceous glands in the skin and exists on the skin surface as a thin layer covering the skin surface. SSL contains RNA expressed in skin cells. The skin from which SSL is collected may be any part of the body, such as the head, face, neck, trunk, limbs, etc., and preferably is a part where sebum is secreted in large amounts, such as the skin of the face.

[0014] The method for collecting a biological sample can be selected appropriately depending on the type of biological sample. For example, any means used for recovering or removing SSL from the skin can be used to collect SSL from the subject's skin. Preferably, an SSL-absorbent material, an SSL-adhesive material, or an instrument for scraping SSL from the skin can be used. The SSL-absorbent material or SSL-adhesive material can be any material that has affinity for SSL, including polypropylene, pulp, etc. More specific examples of means for collecting SSL from the skin include absorbing SSL into a sheet-like material such as oil blotting paper or oil blotting film, adhering SSL to a glass plate or tape, or scraping SSL off with a spatula, scraper, or the like. To improve SSL adsorption, an SSL-absorbent material pre-soaked with a highly lipid-soluble solvent may be used.

[0015] The method for extracting genes from a biological sample can be selected appropriately depending on the biological sample and the gene to be extracted. For example, RNA can be extracted from SSL using methods commonly used for extracting or purifying RNA from biological samples, such as the acid guanidinium thiocyanate-phenol-chloroform extraction (AGPC) method, the spin column method, or a method using magnetic particles.

[0016] The method for measuring the expression level of a gene or its expression product from a biological sample can be selected appropriately depending on the target of measurement. For example, when RNA, cDNA, or DNA is used as the target, the method for measuring the expression level can be selected from nucleic acid amplification methods such as PCR using DNA that hybridizes to these as primers, real-time RT-PCR, multiplex PCR, SmartAmp, and LAMP, hybridization methods using nucleic acids that hybridize to these as probes (DNA chips, DNA microarrays, dot blot hybridization, slot blot hybridization, Northern blot hybridization, etc.), methods for determining base sequences (sequencing), or combinations of these. For example, when RNA is used as the target of measurement of the expression level of a gene or its expression product, the expression level of RNA may be analyzed, but preferably, the RNA is converted into cDNA by reverse transcription, and the expression level of the cDNA or its amplification product is then measured.

[0017] Furthermore, for example, when the target is a gene translation product (protein), a molecule that interacts with the protein, a molecule that interacts with RNA, or a molecule that interacts with DNA, methods for measuring the expression level include protein chip analysis, immunoassays (e.g., ELISA, etc.), mass spectrometry (e.g., LC-MS / MS, MALDI-TOF / MS), the one-hybrid method (PNAS 100, 12271-12276 (2003)), and the two-hybrid method (Biol. Reprod. 58, 302-311 (1998)), and these can be selected appropriately depending on the target.

[0018] In the following description, gene names are based on the official symbols listed in NCBI ([www.ncbi.nlm.nih.gov / ]). Furthermore, genes that can serve as diagnostic markers in each embodiment of the present invention also include genes having a base sequence substantially identical to the base sequence of the DNA constituting the gene, as long as they have the function of determining the dieting behavior of a subject. Here, a substantially identical base sequence means, for example, that when searched using the homology calculation algorithm NCBI BLAST under the conditions of expectation value = 10; gaps allowed; filtering = ON; match score = 1; mismatch score = -3, the base sequence has 90% or more identity with the base sequence of the DNA constituting the gene, preferably 95% or more, and even more preferably 98% or more identity.

[0019] In one embodiment of the present invention, the term "subject" refers to a person who is the subject of the assessment of dieting behavior according to the present invention. Furthermore, "subjects who are the subject of the assessment of dieting behavior" include those who have voluntarily undertaken dieting behavior, and those who have been instructed to undertake dieting behavior by a weight loss instructor (such as a medical professional or a trainer at a health promotion facility), and there are no limitations on whether or not they have actually been on a dieting behavior, or on the spontaneity of the dieting behavior. The gender of the subject is not particularly limited. Furthermore, the age group of the subject is not particularly limited, but from the perspective of improving prediction accuracy, it is preferable that the subject be an adult, preferably between 18 and 80 years old, and more preferably between 20 and 60 years old.

[0020] The obesity level of the subject is not particularly limited, but preferably the subject is, for example, of normal weight and / or of high obesity. A normal weight subject is, for example, a subject with a Body Mass Index (BMI) of 18.50 or more and less than 25.00. A high obesity subject is, for example, a subject with a BMI of 25.00 or more. An example of the subject of the present invention is a subject with a BMI of 23.00 or more. In addition to the above-mentioned BMI, "obesity level" can be evaluated using, for example, visceral fat mass, visceral fat area, subcutaneous fat mass, body fat mass, body fat percentage, waist circumference, abdominal circumference, weight, body mass index (Rohrer index, Kaup index, etc.), obesity index calculated from the body mass index, subcutaneous fat thickness in the triceps, subcutaneous fat thickness in the subscapularis, basal metabolic rate, blood markers correlated with visceral fat mass (cytokines such as adiponectin, leptin, TNFα, and IL-6, inflammatory markers such as CRP, blood lipids such as LDL-cholesterol and triglycerides, liver function markers, blood glucose levels, etc.).

[0021] In one embodiment of the present invention, the term "diet behavior" refers to behavior for weight loss. In one embodiment of the present invention, the diet behavior includes energy intake restriction behavior and energy consumption promotion behavior.

[0022] In one embodiment of the present invention, "energy intake restriction behavior" refers to behavior that restricts energy intake through diet. Examples of energy intake restriction behavior include at least one behavior selected from the group consisting of reducing carbohydrate intake and intake ratio, reducing lipid intake and intake ratio, increasing protein intake ratio, and increasing dietary fiber intake. The amount of reduction in dietary energy intake is not particularly limited, but can be, for example, a reduction in energy intake aimed at a target weight (a 1-10% reduction, preferably a 1-5% reduction, more preferably a 3% reduction from current weight).

[0023] In one embodiment of the present invention, the energy intake restriction behavior is preferably a behavior that is continuously performed. The period for continuously performing the energy intake restriction behavior is preferably one week or more, more preferably ten days or more, and even more preferably one month or more. Furthermore, during this period, the frequency of consciously consuming a diet that restricts energy intake is preferably one or more times a day, and more preferably every meal.

[0024] In one embodiment of the present invention, "energy consumption promoting behavior" refers to behavior that promotes energy consumption by increasing the amount of exercise. "Amount of exercise" refers to the amount of exercise determined based on the intensity, duration, and frequency of the exercise. "Exercise" refers to a behavior (physical activity) that consumes more energy than a state of rest, and is intentionally performed for the purpose of weight loss and is continuous. Specifically, examples of energy consumption promoting behavior in a subject include specific types of exercise such as sports, training, and aerobic exercise, which are performed in addition to the subject's current physical activity, as well as increasing the intensity, duration, and frequency of physical activity such as work or housework. More specific examples of energy expenditure-promoting behaviors include, but are not limited to, exercise that increases energy expenditure to achieve a target weight (a 1-10% reduction from current weight, preferably a 1-5% reduction, and more preferably a 3% reduction), as well as moderate-intensity (3.0-6.0 METs) physical activity (e.g., brisk walking, dancing, mowing the lawn, etc.) for a predetermined time per week (e.g., at least 150-300 minutes), high-intensity (6.0 METs or more) physical activity (e.g., running, swimming, climbing stairs, etc.) for a predetermined time per week (e.g., at least 75-150 minutes), and prescribed strength training (e.g., 15 minutes of multiple strength training exercises such as squats) performed a predetermined number of times per week (e.g., about three times).

[0025] In one embodiment of the present invention, the energy consumption promotion behavior is preferably a behavior that is performed continuously. The period for performing the energy consumption promotion behavior continuously is preferably one week or more, more preferably ten days or more, and even more preferably one month or more. Furthermore, during this period, the frequency with which the energy consumption promotion behavior is consciously performed is preferably three days or more per week, and more preferably every day.

[0026] In one embodiment of the present invention, the target period for assessment is the period up to the time of collection of the biological sample that is the subject's assessment of the implementation of the dieting behavior. For example, if the biological sample is collected at a single point in time, the target period for assessment is the period up to the time of collection of the biological sample. In this case, the start of the target period for assessment is not particularly limited, and can be, for example, the start of the subject's subjective dieting behavior or the start of the dieting behavior instructed by a weight loss instructor. Furthermore, if the start of these dieting behaviors is unknown, the target period for assessment can be set to a typical diet period (preferably one week or more, preferably ten days or more, more preferably one month or more). Alternatively, if the biological sample is collected at two or more points in time, the target period for assessment is set to the time from the earliest biological sample collection to the latest biological sample collection. Note that the "target period for assessment" is the target period for processing in the assessment method described below, and may be, for example, a portion of a diet program in which the subject participates.

[0027] Specific embodiments of the present invention will be described below: The embodiments of the present invention are not intended to be used for medical purposes on humans, and do not include any therapeutic or diagnostic procedures for medical purposes.

[0028] [First embodiment] The determination method according to the first embodiment of the present invention includes a determination step of determining a diet behavior selected from energy intake restriction behavior and energy consumption promotion behavior performed by a subject within a period to be determined up to the time of collection of the biological sample, based on the expression level of at least one gene or its expression product selected from the group consisting of intake restriction variable genes and consumption promotion variable genes derived from the biological sample of the subject whose diet behavior is to be determined.

[0029] In this embodiment, the intake restriction variable gene is at least one gene whose expression level varies with energy intake restriction behavior but does not vary with energy consumption-promoting behavior. In this embodiment, the intake restriction variable gene functions as a determination marker for determining whether a subject is engaging in energy intake restriction behavior. Furthermore, a "gene whose expression level varies with energy intake restriction behavior but does not vary with energy consumption-promoting behavior" refers to a gene whose expression level or its expression product is shown to increase or decrease statistically significantly in gene expression data before and after intervention in a subject population in which only energy intake restriction behavior is intervened, and whose expression level or its expression product is shown not to change statistically significantly in gene expression data before and after intervention in a subject population in which only energy consumption-promoting behavior is intervened.

[0030] In this embodiment, the intake restriction fluctuating gene is at least one gene selected from the group consisting of intake restriction increasing genes whose expression levels increase due to energy intake restriction behavior and intake restriction decreasing genes whose expression levels decrease due to energy intake restriction behavior. Specifically, intake restriction increasing genes are genes whose expression levels have been shown to significantly increase before and after intervention based on data from a subject population in which only the above-mentioned energy intake restriction behavior was intervened. Similarly, intake restriction decreasing genes are genes whose expression levels have been shown to significantly decrease before and after intervention based on data from a subject population in which only the above-mentioned energy intake restriction behavior was intervened.

[0031] In this embodiment, the gene that increases intake restriction preferably consists of at least one gene selected from the group consisting of 34 genes: KIAA0146, FBXW4, LRP11, TCEA3, ANXA9, SPINK7, DGCR6L, CA13, THOC3, MYL9, RPIA, TPRG1, FAM83H, AGR2, DANCR, FAM108B1, RAB11FIP5, RPL13, TSNARE1, TFAP2A, EPHA2, NDUFA4L2, JMJD8, RASD2, PMM1, SNX21, NCK2, ALOX12B, RAB38, BSCL2, HRAS, CNKSR3, GLE1, and CDKN1C.

[0032] In this embodiment, the intake restriction reducing gene preferably consists of at least one gene selected from the group consisting of 23 genes: CCDC82, LOC646214, TMEM14E, C5orf15, CCNL1, NRARP, LOC145474, IFIH1, RNF2, CRLF2, ITPRIPL2, LOC100303749, PRPF40A, MTSS1, CLNS1A, MEF2A, RB1, ANKRD44, ZNF292, NXF1, ARID4B, ZCRB1, and LPAR6.

[0033] In the Examples described below, each of the 57 intake restriction variable genes is a gene that was shown to have a significantly increased or decreased mRNA expression level in the data before and after the dietary intervention in the homogeneous dietary intervention subject group, a significantly unchanged mRNA expression level in the data before and after the exercise intervention in the homogeneous exercise intervention subject group, and a significantly increased or decreased mRNA expression level in the data before and after the exercise intervention in the homogeneous exercise intervention subject group, based on data from a group of subjects who underwent a dietary management instruction program (dietary intervention) and were certified to have complied with the instructions in the program (hereinafter referred to as the "homogeneous dietary intervention subject group"), among those who underwent an instruction program for increasing physical activity (exercise intervention), and a significantly increased or decreased mRNA expression level in the data before and after the exercise intervention in the homogeneous exercise intervention subject group, and a significantly increased or decreased RNA expression level in the ratio before and after the dietary intervention, and a significantly increased or decreased RNA expression level in the ratio before and after the dietary intervention in the homogeneous exercise intervention subject group. Therefore, each of the 57 intake restriction variable genes can function as a marker for determining whether or not energy intake restriction behavior equivalent to dietary intervention is being performed.

[0034] In this embodiment, the consumption-promoting variable gene is at least one gene whose expression level does not vary with energy intake restriction behavior, but whose expression level varies with energy consumption-promoting behavior. In this embodiment, the consumption-promoting variable gene functions as a determination marker for determining whether a subject is engaging in energy consumption-promoting behavior. Furthermore, a "gene whose expression level does not vary with energy intake restriction behavior, but whose expression level varies with energy consumption-promoting behavior" refers to a gene whose expression level or its expression product increases or decreases statistically significantly in gene expression data from a subject group treated with only energy consumption-promoting behavior, and whose expression level does not change statistically significantly in gene expression data from a subject group treated with only energy intake restriction behavior before and after the intervention.

[0035] In this embodiment, the consumption promotion variable gene is at least one gene selected from the group consisting of a consumption promotion increasing gene whose expression level increases due to an energy consumption promoting behavior and a consumption promotion decreasing gene whose expression level decreases due to an energy consumption promoting behavior. Specifically, the consumption promotion increasing gene is a gene whose expression level has been shown to increase significantly before and after the intervention based on data from a subject group in which only the above energy consumption promoting behavior was intervened. Similarly, the consumption promotion decreasing gene is a gene whose expression level has been shown to decrease significantly before and after the intervention based on data from a subject group in which only the above energy consumption promoting behavior was intervened.

[0036] In this embodiment, the consumption-promoting gene is preferably at least one gene selected from the group consisting of 41 genes: SCEL, KIAA0284, BARX2, S100A2, KRTAP1-5, CAMK2N1, KRT13, PIP, AIF1L, GBAP1, KRTAP3-3, PRKCZ, EVPL, WBSCR22, ARG1, KRTAP5-10, SEPP1, PLK2, KRT6C, GSDMC, PLEKHG1, CDC42BPG, MAST4, GOLGA7B, CGA, SDR16C5, ARHGAP23, ATP6V0A4, RERE, KLC3, CGN, RAPGEF5, OSBPL1A, MAL, FMN1, CXADR, OCLN, COQ9, NIPAL4, BAIAP2L1, and SH3GL3.

[0037] In this embodiment, the consumption-promoting reducing gene preferably comprises at least one gene selected from the group consisting of the five genes LOC100506888, OR11H12, LOC349196, USP17L6P, and USP17L5.

[0038] In the Examples described below, each of the 46 consumption-promoting variable genes is a gene whose mRNA expression level was shown to be significantly increased or decreased in data before and after exercise intervention in a homogeneous exercise-intervention subject group, whose mRNA expression level was not significantly changed in data before and after dietary intervention in a homogeneous dietary intervention subject group, and whose RNA expression level ratio before and after the exercise intervention was more than 2-fold or less than 1 / 2. Therefore, each of the 46 consumption-promoting variable genes can function as a marker for determining whether or not an energy consumption-promoting behavior equivalent to exercise intervention has been performed.

[0039] Furthermore, the expression levels of the intake restriction variable gene and / or consumption promotion variable gene, or their expression products, are preferably derived from a non-invasively collected biological sample such as SSL, which allows for easier collection of the biological sample and reduces the burden on the subject compared to using an invasively collected biological sample such as blood.

[0040] In the determination step of this embodiment, the diet behavior performed by the subject during the determination period can be determined based on the expression level of at least one gene selected from the group consisting of intake restriction variable genes and consumption promotion variable genes, or its expression product, in the following manner. For example, the expression level can be used to determine whether the condition corresponding to the performance of at least one of energy intake restriction behavior or energy consumption promotion behavior is met. Alternatively, as described below, a determination model for determining the diet behavior performed based on the expression level can be used, and the expression level derived from the subject's biological sample can be applied to this determination model to determine the diet behavior performed by the subject. These determination processes can be performed, for example, by a computer.

[0041] The derived judgment results include, for example, "energy intake restriction behavior was performed," "energy consumption promotion behavior was performed," "energy intake restriction behavior and energy consumption promotion behavior were performed," "neither energy intake restriction behavior nor energy consumption promotion behavior was performed," and "either energy intake restriction behavior or energy consumption promotion behavior was performed predominantly."

[0042] In the present embodiment, when determining whether or not a condition corresponding to the execution of at least one of the energy intake restriction behavior and the energy consumption promotion behavior is satisfied, the determination step includes a condition satisfaction determination step and a behavior determination step. This aspect will be specifically described below.

[0043] In the condition fulfillment determination step of this embodiment, it is determined whether the expression level of at least one gene selected from the group consisting of intake restriction variable genes and consumption promotion variable genes derived from the subject's biological sample or its expression product, or a change index representing a change in the expression level derived from the subject's biological sample during the period to be determined, satisfies a predetermined condition.

[0044] The determination marker used in this step may be at least one intake restriction variable gene, or at least one consumption promotion variable gene, or may include both intake restriction variable genes and consumption promotion variable genes. In particular, the determination marker used in this step is preferably at least one gene selected from the above-mentioned 34 intake restriction increasing genes, 23 intake restriction decreasing genes, 41 consumption promotion increasing genes, and 5 consumption promotion decreasing genes. Furthermore, the number of determination markers used may be one or more, but from the viewpoint of improving the accuracy of determination, it is preferable to use more than one.

[0045] The value indicating the expression level used in this step may specifically take the following values. For example, values ​​indicating the expression level when analyzing the expression levels of multiple genes by sequencing include a read count value, which is expression level data, an RPM value obtained by correcting the read count value for the difference in the total number of reads between samples, a value obtained by converting the RPM value to a base 2 logarithm (Log2RPM value), a base 2 logarithm obtained by adding an integer 1 to the RPM value (Log2(RPM+1) value), a count value corrected using DESeq2 (Normalized count value), and a base 2 logarithm obtained by adding an integer 1 (Log2(count+1) value). Other examples of values ​​indicating the expression level may include values ​​calculated using common quantitative values ​​in RNA-seq, such as fragments per kilobase of exon per million reads mapped (FPKM), reads per kilobase of exon per million reads mapped (RPKM), and transcripts per million (TPM). Other examples of values ​​indicating the expression level include signal values ​​obtained by microarray analysis and their corrected values. When analyzing the expression level of only a specific gene by RT-PCR or the like, the value indicating the expression level may be a value calculated by a method in which the expression level of the gene to be measured is converted to a relative expression level based on the expression level of a housekeeping gene (relative quantification), or by a method in which the absolute copy number is quantified using a plasmid containing the region of the gene to be measured (absolute quantification). Alternatively, the copy number obtained by digital PCR may be used as the expression level.

[0046] In this step, a change index representing a change in the expression level of a gene or its expression product derived from a subject's biological sample during a period of evaluation refers to an index representing a change between the expression level of the gene or its expression product derived from the subject's biological sample at the start of the period of evaluation and the expression level of the gene or its expression product derived from the subject's biological sample at the end of the period of evaluation. Specific examples of such change indexes include the amount of change or rate of change indicating the difference in the expression level between the start and end of the period of evaluation, the ratio of the expression level at the start of the period of evaluation to the expression level at the end of the period of evaluation (expression amount ratio), and indices indicating the range thereof. The "rate of change in the value indicating the expression level" refers to the ratio of the difference (amount of change) to the value at the start of the period of evaluation. The numerator and denominator in the above "expression level ratio" are not limited, and may be, for example, the ratio of the expression level at the end of the period to be determined to the expression level at the start of the period to be determined (expression level at the end of the period to be determined / expression level at the start of the period to be determined), or the ratio of the expression level at the start of the period to the expression level at the end of the period to be determined (expression level at the start of the period to be determined / expression level at the end of the period to be determined).

[0047] The predetermined conditions are conditions corresponding to the implementation of energy intake restriction behavior and / or energy consumption promoting behavior. Examples of processing using the predetermined conditions in this step include processing in which an intake restriction variable gene is used as a determination marker to determine whether the expression level or change indicator of the intake restriction variable gene satisfies the condition corresponding to the implementation of energy intake restriction behavior. Another example includes processing in which a consumption promotion variable gene is used as a determination marker to determine whether the expression level or change indicator of the consumption promotion variable gene satisfies the condition corresponding to the implementation of energy consumption promoting behavior. Yet another example includes processing in which both an intake restriction variable gene and a consumption promotion variable gene are used as determination markers to determine whether the expression level or change indicator of the intake restriction variable gene satisfies the condition corresponding to the implementation of energy intake restriction behavior, and whether the expression level or change indicator of the consumption promotion variable gene satisfies the condition corresponding to the implementation of energy consumption promoting behavior.

[0048] The predetermined conditions may include conditions set for each gene used as a determination marker. The conditions set for each intake restriction variable gene are conditions set in response to the implementation of energy intake restriction behavior, and the conditions set for each consumption promotion variable gene are conditions set in response to the implementation of energy consumption promotion behavior. Specific examples of such conditions include the range of values ​​that can be assumed for the expression level and / or change index of each gene or its expression product, and values ​​and ranges thereof calculated by substituting a numerical value indicating the expression level and / or its change index into a predetermined mathematical formula. In this example, when two or more genes are used as determination markers, the predetermined conditions may further include, in addition to the conditions set for each gene, a condition regarding the number or proportion of intake restriction variable genes that satisfy the condition, or a condition regarding the number or proportion of consumption promotion variable genes that satisfy the condition. As another example, when two or more genes are used as determination markers, the predetermined conditions may be a value or range thereof calculated by substituting a value indicating the expression level and / or change index corresponding to each of the multiple genes into a predetermined mathematical formula.

[0049] In the behavior determination step of this embodiment, a diet behavior selected from energy intake restriction behavior and energy consumption promotion behavior performed by the subject during the determination period is determined based on the determination result in the condition satisfaction determination step.

[0050] The behavior determination step in this embodiment may take the following forms. For example, this step may determine whether or not an energy intake restriction behavior has been performed based on the determination result of a condition satisfaction determination step using an intake restriction variation gene as a determination marker. As another example, this step may determine whether or not an energy consumption promotion behavior has been performed based on the determination result of a condition satisfaction determination step using a consumption promotion variation gene as a determination marker. As another example, the determination may determine whether or not at least one of an energy intake restriction behavior or an energy consumption promotion behavior has been performed based on the determination result of a condition satisfaction determination step using both an intake restriction variation gene and a consumption promotion variation gene as determination markers. As another example, the determination may determine whether an energy intake restriction behavior or an energy consumption promotion behavior has been performed predominantly based on the determination result of a condition satisfaction determination step using both an intake restriction variation gene and a consumption promotion variation gene as determination markers.

[0051] As described above, the method of this embodiment uses at least one gene selected from the group consisting of intake restriction variable genes and consumption promotion variable genes derived from the biological sample of the subject whose diet behavior is to be assessed as a marker to assess the diet behavior performed by the subject during the assessment period. This provides objective information for assessing the diet behavior actually performed by the subject, and the assessment results can be used for providing weight loss guidance, lifestyle guidance, etc. to the subject.

[0052] Taking advantage of these advantages, the assessment method of this embodiment can be used, for example, by a weight loss instructor to assess the diet behavior actually performed by a subject, or by a subject who wishes to lose weight to objectively assess his or her own diet behavior.

[0053] Hereinafter, embodiments corresponding to the aspects described in the above embodiment will be described. Note that in the following embodiments, descriptions of configurations similar to or corresponding to those in the first embodiment will be omitted as appropriate.

[0054] [Second embodiment] The method according to the second embodiment of the present invention includes a step of determining whether or not a subject has engaged in energy intake restriction behavior during a period of time to be determined, based on the expression level of at least one gene selected from the group consisting of intake restriction fluctuating genes or its expression product, derived from a biological sample from the subject. The intake restriction fluctuating gene, which is the determination marker used in this step, can be, for example, at least one selected from the 34 intake restriction increasing genes and the 23 intake restriction decreasing genes described above.

[0055] As in the first embodiment, the determination step of this embodiment preferably includes, for example, a condition satisfaction determination step and a behavior determination step.

[0056] In this embodiment, the condition fulfillment determination step determines whether the expression level of at least one gene selected from the group consisting of intake restriction variable genes or its expression product derived from the subject's biological sample, or the change index representing the change in the expression level derived from the subject's biological sample during the period to be determined, fulfills a predetermined condition corresponding to the implementation of energy intake restriction behavior.

[0057] The predetermined conditions corresponding to the implementation of the energy intake restriction behavior can be the various conditions described in the first embodiment. For example, when the predetermined conditions include conditions set for each intake restriction variable gene, the predetermined conditions can be the range of values ​​that can be taken by the expression level and / or change index of each intake restriction gene or its expression product, or a value or range calculated by substituting a numerical value indicating the expression level and / or change index into a predetermined formula. In this example, when there are two or more intake restriction variable genes, the predetermined conditions can further include, in addition to the conditions set for each gene, a condition regarding the number or proportion of intake restriction variable genes that satisfy the condition. As another example, when there are two or more intake restriction variable genes, the predetermined conditions can be a value or range calculated by substituting a value indicating the expression level and / or change index corresponding to each of the multiple genes into a predetermined formula.

[0058] The predetermined conditions may be set for each intake restriction fluctuating gene, and may include, for example, a condition that includes a reference value set for each gene as an endpoint of a numerical range. For example, if the intake restriction fluctuating gene includes an intake restriction increasing gene, the predetermined conditions may include a condition that the expression level or the change index of the intake restriction increasing gene is equal to or greater than a reference value set for each gene. Alternatively, if the intake restriction fluctuating gene includes an intake restriction decreasing gene, the predetermined conditions may include a condition that the expression level or the change index of the intake restriction decreasing gene is equal to or less than a reference value set for each gene, or less than the reference value.

[0059] The reference value can be set, for example, using a numerical value that represents a clear increase or decrease in the expression level during the period to be determined (e.g., the numerical ratio of the expression level at the start and end of the period to be determined), or a statistical value (mean, median, quartile, 95% confidence interval, etc.) of the expression level of each gene or its expression product or the above-mentioned change indicator calculated from data on a group of subjects who engaged in energy intake restriction behavior.

[0060] In the present embodiment, in the behavior determination step, if it is determined that the above-mentioned predetermined condition is satisfied, it is determined that the subject has performed an energy intake restriction behavior within the determination target period.

[0061] As described above, according to this embodiment, it is possible to determine whether or not energy intake restriction behavior has actually been performed using intake restriction fluctuating genes.

[0062] [Third embodiment] The determination method according to the third embodiment of the present invention includes a determination step of determining whether or not the subject performed an energy consumption-promoting behavior during a period of time to be determined based on the expression level of at least one gene selected from the group consisting of consumption-promoting variable genes or its expression product, which is derived from a biological sample of the subject. The consumption-promoting variable gene, which is the determination marker used in this step, can be, for example, at least one selected from the above-mentioned 41 consumption-promoting increasing genes and 5 consumption-promoting decreasing genes.

[0063] Specifically, similarly to the first embodiment, the determining step of this embodiment preferably includes a condition satisfaction determining step and a behavior determining step.

[0064] In this embodiment, in the condition fulfillment determination step, it is determined whether the change index representing the change in the expression level of at least one gene selected from the group consisting of consumption-promoting variable genes derived from the subject's biological sample or its expression product, or the expression level derived from the subject's biological sample, during the period to be determined, fulfills a predetermined condition corresponding to the implementation of energy consumption-promoting behavior.

[0065] The predetermined conditions corresponding to the implementation of the energy consumption promotion behavior can be the various conditions described in the first embodiment. For example, when the predetermined conditions include conditions set for each consumption promotion variable gene, the predetermined conditions may include the range of values ​​that can be assumed for the expression level and / or change index of each consumption promotion gene or its expression product, or a value or range calculated by substituting a value indicating the expression level and / or change index into a predetermined formula. In this example, when there are two or more consumption promotion variable genes, the predetermined conditions may further include, in addition to the conditions set for each gene, a condition regarding the number or proportion of consumption promotion variable genes that satisfy the condition. As another example, when there are two or more consumption promotion variable genes, the predetermined conditions may be a value or range calculated by substituting a value indicating the expression level and / or change index corresponding to each of the multiple genes into a predetermined formula.

[0066] The predetermined conditions may be set for each consumption promotion variable gene, and may include, for example, a condition that includes a reference value set for each gene as an endpoint of a numerical range. For example, if the consumption promotion variable gene includes a consumption promotion increasing gene, the predetermined conditions may include a condition that the expression level or change index of the consumption promotion increasing gene is equal to or greater than a reference value set for each gene. Alternatively, if the consumption promotion variable gene includes a consumption promotion decreasing gene, the predetermined conditions may include a condition that the expression level or change index of the consumption promotion decreasing gene is equal to or less than a reference value set for each gene, or less than the reference value.

[0067] The above-mentioned reference value can be set, for example, using a numerical value that represents a clear increase or decrease in the expression level during the period to be determined (for example, the numerical ratio of the expression level at the start and end of the period to be determined), or a statistical value (mean, median, quartile, 95% confidence interval, etc.) of the expression level of each gene or its expression product or the above-mentioned change indicator calculated from data on a group of subjects who engaged in energy consumption-promoting behavior.

[0068] In the present embodiment, in the behavior determination step, if it is determined that the above-mentioned predetermined condition is satisfied, it is determined that the subject has performed the energy consumption promoting behavior within the determination target period.

[0069] As described above, according to this embodiment, it is possible to determine whether or not an energy consumption promoting behavior has actually been performed using consumption promotion variable genes.

[0070] [Fourth embodiment] (Judgment process) Furthermore, the determination method according to a fourth embodiment of the present invention includes a determination step of determining whether a subject has engaged in a dieting behavior selected from energy intake restriction behavior and energy consumption promotion behavior during a period of time to be determined, based on the expression level of at least one gene selected from the group consisting of intake restriction variable genes or its expression product and the expression level of at least one gene selected from the group consisting of consumption promotion variable genes or its expression product. In this embodiment, the determination of dieting behavior is characterized by determining whether the subject engaged in at least one of energy intake restriction behavior and energy consumption promotion behavior during a period of time to be determined. The intake restriction variable gene used as a determination marker in this step can be, for example, at least one selected from the 34 intake restriction increasing genes and the 23 intake restriction decreasing genes described above. The consumption promotion variable gene used as a determination marker in this step can be, for example, at least one selected from the 41 consumption promotion increasing genes and the 5 consumption promotion decreasing genes described above.

[0071] The determination process of this embodiment includes, for example, a first condition satisfaction determination process, a second condition satisfaction determination process, and a behavior determination process. The first condition satisfaction process is the same as the condition satisfaction process described in the second embodiment, and the second condition satisfaction process is the same as the condition satisfaction process described in the third embodiment. Note that the order of the first condition satisfaction determination process and the second condition satisfaction determination process may be any order.

[0072] In the first condition satisfaction determination step of this embodiment, it is determined whether the expression level of at least one gene selected from the group consisting of intake restriction variable genes or its expression product derived from the subject's biological sample, or the change index representing the change in the expression level derived from the subject's biological sample during the determination period, satisfies the first condition corresponding to the implementation of energy intake restriction behavior. The first condition can be various conditions similar to the "predetermined condition" in the condition satisfaction step of the second embodiment.

[0073] In the second condition satisfaction determination step of this embodiment, it is determined whether the expression level of at least one gene selected from the group consisting of consumption-promoting variable genes or its expression product derived from the subject's biological sample, or the change index representing the change in the expression level derived from the subject's biological sample during the determination period, satisfies the second condition corresponding to the implementation of energy consumption-promoting behavior. The second condition can be various conditions similar to the "predetermined condition" in the condition satisfaction step of the third embodiment.

[0074] In the behavior determination process of this embodiment, it is determined whether or not the subject has performed at least one of an energy intake restriction behavior or an energy consumption promotion behavior within the determination period based on the determination results of the first condition fulfillment determination process and the second condition fulfillment determination process.

[0075] For example, in the behavior determination step of this embodiment, if it is determined that the first condition is satisfied in the first condition satisfaction determination step, it is determined that the energy intake restriction behavior has been performed, and if it is determined that the second condition is satisfied in the second condition satisfaction determination step, it is determined that the energy consumption promotion behavior has been performed. Therefore, the determination result can take four patterns: "both the energy intake restriction behavior and the energy consumption promotion behavior have been performed," "the energy intake restriction behavior has been performed but the energy consumption promotion behavior has not been performed," "the energy consumption promotion behavior has been performed but the energy intake restriction behavior has not been performed," and "neither the energy intake restriction behavior nor the energy consumption promotion behavior has been performed."

[0076] As described above, according to this embodiment, it is possible to determine whether or not energy intake restriction behavior and / or energy consumption promotion behavior has been performed using both intake restriction fluctuating genes and consumption promotion fluctuating genes.

[0077] [Fifth embodiment] The determination method of the fifth embodiment of the present invention, like the fourth embodiment, performs a determination process using both intake restriction fluctuating genes and consumption promoting genes, but differs from the fourth embodiment in that it uses multiple intake restriction fluctuating genes and multiple consumption promoting genes to determine whether energy intake restriction behavior or energy consumption promoting behavior was predominantly performed within the determination period.

[0078] Specifically, the determination step of this embodiment includes a first condition satisfaction determination step, a second condition satisfaction determination step, and a behavior determination step.

[0079] In this embodiment, in the step of determining whether the first condition is satisfied, it is determined whether the expression levels of multiple genes or their expression products selected from a group consisting of intake restriction variable genes derived from the subject's biological sample at the end of the period to be determined, or the change index representing the change in the expression level derived from the subject's biological sample between the start and end of the period to be determined, satisfy the first condition set for each gene.

[0080] In this step, the first condition is a condition set for each of the above genes, and may be any of the various conditions set for each gene described in Embodiment 2. Specific examples of the first condition include the range of values ​​that can be taken by the expression level and / or its change index of each gene or its expression product, and a value or range thereof calculated by substituting a numerical value indicating the expression level and / or its change index into a predetermined mathematical formula.

[0081] In this embodiment, in the second condition satisfaction determination step, it is determined whether the change index representing the change in the expression levels of multiple genes or their expression products selected from a group consisting of consumption-promoting variable genes derived from the subject's biological sample, or the expression levels derived from the subject's biological sample, during the period to be determined, satisfies the second condition set for each gene.

[0082] In this step, the second condition is a condition set for each of the above genes, and may be any of the various conditions set for each gene described in Embodiment 3. Specific examples of the second condition include the range of values ​​that can be taken by the expression level and / or its change index of each gene or its expression product, and a value or range thereof calculated by substituting a numerical value indicating the expression level and / or its change index into a predetermined mathematical formula.

[0083] In the behavioral assessment process of this embodiment, first, a first condition satisfaction degree indicating the number or proportion of intake restriction variable genes that satisfy the first condition in the first condition satisfaction assessment process, and a second condition satisfaction degree indicating the number or proportion of consumption promotion variable genes that satisfy the second condition in the second condition satisfaction assessment process are calculated.

[0084] "The proportion of intake restriction variable genes that satisfy the first condition in the first condition satisfaction determination step" means the proportion of the number of intake restriction variable genes that satisfy the first condition to the number of all intake restriction variable genes used in the first condition satisfaction determination step. Similarly, "the proportion of consumption promotion variable genes that satisfy the second condition in the second condition satisfaction determination step" means the proportion of the number of consumption promotion variable genes that satisfy the second condition to the number of all consumption promotion variable genes used in the second condition satisfaction determination step.

[0085] Next, in the behavior determination step of this embodiment, the first condition satisfaction degree and the second condition satisfaction degree are compared. If the first condition satisfaction degree is greater than the second condition satisfaction degree, it is determined that the energy intake restriction behavior was performed more preferentially than the energy consumption promotion behavior. If the first condition satisfaction degree is less than the second condition satisfaction degree, it is determined that the energy consumption promotion behavior was performed more preferentially than the energy intake restriction behavior. Note that in this embodiment, if the first condition satisfaction degree and the second condition satisfaction degree are determined to be the same, a determination result may be derived that both behaviors were performed equally, or whether these behaviors were performed may be determined depending on the values ​​of these satisfaction degrees.

[0086] In this embodiment, "energy intake restriction behavior was predominantly performed during the determination period" means that energy intake restriction behavior was more actively performed than energy consumption promotion behavior during the determination period. Similarly, "energy consumption promotion behavior was predominantly performed during the determination period" means that energy consumption promotion behavior was more actively performed than energy intake restriction behavior during the determination period.

[0087] In this way, in this embodiment, by using both intake restriction fluctuation genes and consumption promotion fluctuation genes, it is possible to determine whether energy intake restriction behavior or energy consumption promotion behavior was more prevalent during the period to be determined.

[0088] [Other embodiments] Although the embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments, and it goes without saying that various modifications can be made within the scope of the gist of the present invention.

[0089] The determination method according to another embodiment of the present invention may further include an acquisition step of acquiring the expression level of at least one gene or its expression product selected from the group consisting of intake restriction variable genes and consumption promotion variable genes, derived from a biological sample from a subject. In the acquisition step of this embodiment, "acquiring the expression level of a gene or its expression product" refers to acquiring information on the expression level of a gene or its expression product, and includes, for example, at least one of measuring the expression level of a gene or its expression product and acquiring information on the measurement results, or acquiring expression level information from another device, etc. Examples of ways of acquiring expression level information from a device, etc. include, for example, receiving expression level information from a computer, etc. via wireless or wired connections, and reading expression level information from a storage medium, etc., that stores expression level information.

[0090] Furthermore, in the above-mentioned acquisition step, the expression level information is preferably acquired in association with information on the subject from whom the biological sample was collected and information indicating the timing of collection of the biological sample. The subject information includes information to identify the subject (e.g., subject ID, name (nickname), email address, etc.), attribute information of the subject (e.g., gender, age (generation), occupation, address (residential area), information on facilities and instructors that provide weight loss guidance, etc.). The information indicating the timing of collection of the biological sample includes the date of collection of the biological sample.

[0091] In the above example, the determination step includes a condition satisfaction determination step and a behavior determination step, but is not limited thereto. For example, in the determination step, the expression level of at least one gene selected from the group consisting of intake restriction variable genes and consumption promotion variable genes derived from the subject's biological sample, or its expression product, may be applied to a determination model that determines diet behaviors selected from energy intake restriction behaviors and energy consumption promotion behaviors based on the expression level, thereby determining the diet behaviors performed by the subject during the determination period. The algorithm used in such a determination model is not particularly limited, and examples include logistic regression, support vector machine (SVM), decision tree, random forest, neural network, etc.

[0092] Another embodiment of the present invention provides a determination device that executes the above-described determination method. The determination device includes a control unit that acquires the expression level of at least one gene selected from the group consisting of intake restriction variable genes and consumption promotion variable genes, or its expression product, derived from a biological sample of a subject whose dieting behavior is to be determined, and determines, based on the acquired expression level, dieting behaviors performed by the subject during the determination period up to the time of collection of the biological sample, selected from energy intake restriction behaviors that restrict energy intake through food and energy consumption promotion behaviors that promote energy consumption by increasing the amount of exercise. The determination device may be any device having a control unit capable of performing the above-described determination process, such as a computer. The control unit may be, for example, a processor such as a CPU.

[0093] As yet another embodiment of the present invention, a determination marker for determining a diet behavior performed by a subject, the diet behavior being selected from an energy intake restriction behavior and an energy consumption promotion behavior, can be provided. The markers for determination are The gene comprises at least one gene selected from the group consisting of intake restriction variable genes whose expression levels fluctuate with energy intake restriction behavior but do not fluctuate with energy consumption promotion behavior, and consumption promotion variable genes whose expression levels do not fluctuate with energy intake restriction behavior but fluctuate with energy consumption promotion behavior. Intake restriction variable genes are An intake restriction-increasing gene whose expression level increases due to energy intake restriction behavior; and an intake restriction-reducing gene whose expression level is reduced by energy intake restriction behavior. Consumption-promoting variable genes are Consumption-promoting genes whose expression levels increase due to energy consumption-promoting behavior; and a consumption-promoting reducing gene whose expression level is reduced by an energy consumption-promoting behavior. The genes that increase intake restriction consist of at least one gene selected from the group consisting of 34 genes: KIAA0146, FBXW4, LRP11, TCEA3, ANXA9, SPINK7, DGCR6L, CA13, THOC3, MYL9, RPIA, TPRG1, FAM83H, AGR2, DANCR, FAM108B1, RAB11FIP5, RPL13, TSNARE1, TFAP2A, EPHA2, NDUFA4L2, JMJD8, RASD2, PMM1, SNX21, NCK2, ALOX12B, RAB38, BSCL2, HRAS, CNKSR3, GLE1, and CDKN1C. The gene that reduces intake restriction consists of at least one gene selected from the group consisting of 23 genes: CCDC82, LOC646214, TMEM14E, C5orf15, CCNL1, NRARP, LOC145474, IFIH1, RNF2, CRLF2, ITPRIPL2, LOC100303749, PRPF40A, MTSS1, CLNS1A, MEF2A, RB1, ANKRD44, ZNF292, NXF1, ARID4B, ZCRB1, and LPAR6. The consumption-promoting elevated gene is at least one gene selected from the group consisting of 41 genes: SCEL, KIAA0284, BARX2, S100A2, KRTAP1-5, CAMK2N1, KRT13, PIP, AIF1L, GBAP1, KRTAP3-3, PRKCZ, EVPL, WBSCR22, ARG1, KRTAP5-10, SEPP1, PLK2, KRT6C, GSDMC, PLEKHG1, CDC42BPG, MAST4, GOLGA7B, CGA, SDR16C5, ARHGAP23, ATP6V0A4, RERE, KLC3, CGN, RAPGEF5, OSBPL1A, MAL, FMN1, CXADR, OCLN, COQ9, NIPAL4, BAIAP2L1, and SH3GL3. The consumption-promoting reducing gene is at least one gene selected from the group consisting of five genes: LOC100506888, OR11H12, LOC349196, USP17L6P, and USP17L5.

[0094] In yet another embodiment of the present invention, there is provided a kit for determining whether a subject has engaged in a dieting behavior selected from the group consisting of an energy intake restriction behavior and an energy consumption promotion behavior, the kit containing a reagent for detecting at least one gene selected from the group consisting of the intake restriction variation gene and the consumption promotion variation gene, or its expression product, from a biological sample of the subject.

[0095] The reagent for detecting the expression level of the gene or its expression product preferably comprises at least one selected from, for example, an oligonucleotide that specifically hybridizes with a nucleic acid derived from the gene and an antibody that recognizes the expression product of the gene. The oligonucleotide comprises at least one selected from, for example, a PCR primer, a hybridization probe, an adapter sequence for sequencing, etc. The antibody may be an antibody that recognizes a protein that is the expression product of the gene.

[0096] In addition to the above-mentioned reagents, the determination kit may also include, for example, reagents for extracting and purifying RNA from the collected SSL, reagents used to detect the expression level of a gene or its expression product, etc. Examples of such reagents include labeling reagents, buffer solutions, chromogenic substrates, secondary antibodies, blocking agents, and control reagents used as positive or negative controls. Furthermore, the determination kit may also include indicators or guidance for detecting the expression level of the gene or its expression product, as well as other instruments necessary for testing.

[0097] In addition to the above-mentioned reagents, the determination kit may also contain tools and reagents necessary for collecting and storing biological samples such as SSLs. For example, tools and reagents necessary for collecting and storing SSLs include oil blotting films for collecting SSLs, reagents for storing the collected SSLs, and storage containers. [Example]

[0098] Example 1: Analysis of gene expression changes before and after weight loss intervention 1) Study participants BMI of 23 kg / m 2 120 adult men (aged 20 to 60 years) who met the eligibility criteria for the study were selected as participants in the dietary intervention group, which will undergo the dietary intervention described below. 113 adult men under the same conditions were selected as participants in the exercise intervention group, which will undergo the exercise intervention described below.

[0099] 2) Collection of lipids on the skin surface Before and after the intervention study described below, RNA-containing skin surface lipids (SSL) were collected from the entire face of the study participants using oil blotting film (5 cm x 8 cm, polypropylene, 3M). The oil blotting film was transferred to a glass vial and stored at -80°C until use for RNA extraction.

[0100] 3) Pre-intervention testing A physical examination was conducted as a pre-intervention test before the intervention test described below. Note that the examination in this test example was conducted with the consent of the test participants.

[0101] Physical examinations were conducted by inviting study participants to a designated venue. Physical examination items included height, weight, body composition (fat mass, lean mass, muscle mass, MQP, determined bone mass, total body water (TBW), total body water percentage (TBW%), body fat percentage, SMI (appendicular muscular index), trunk body fat percentage, trunk body fat mass, trunk lean mass, trunk muscle mass, etc.), visceral fat area, abdominal circumference, blood pressure, pulse, and body temperature. Height was measured using a standard height measuring device. Weight and body composition were measured using a body composition analyzer ("Professional Multi-Frequency Body Composition Analyzer MC-980A-N plus," manufactured by Tanita Corporation). Visceral fat area and abdominal circumference were measured using a Panasonic EW-FA90 visceral fat analyzer (medical device approval number 22500BZX00522000), which was wrapped around the participants' abdomens. Blood pressure and pulse were measured using an upper arm digital blood pressure monitor ("Automatic Blood Pressure Monitor HEM-104", manufactured by Omron Corporation). Body temperature was measured on the forehead using a non-contact thermometer.

[0102] 4) Implementation of dietary intervention Subsequently, the participants in the dietary intervention group underwent a two-month dietary intervention. Specifically, a registered dietitian provided each participant with dietary advice to reduce their calorie intake in order to achieve a target weight (a 3% reduction from their current weight). Dietary advice from the registered dietitian was provided once every two weeks. Each participant was instructed to install an application program (Asuken®) capable of recording their dietary information on their personal information terminal and to record each meal using this application program. In addition, each participant was instructed to measure their daily activity level using a provided activity monitor and not to consciously increase their exercise level. After the two-month dietary intervention, the participants in the dietary intervention group underwent a post-intervention physical examination similar to the pre-intervention examination.

[0103] 5) Selection of subjects for the dietary intervention group Of the 112 participants in the dietary intervention group who completed the two-month dietary intervention, 65 participants who were judged to have achieved adequate dietary management (50% or more of the daily target energy reduction over the entire two-month period) and had not significantly changed their physical activity were selected as a homogeneous group for analysis. Changes in dietary intake were determined using the food records in the application program, and physical activity was determined based on the results of activity measurements.

[0104] 6) Implementation of exercise intervention Meanwhile, the participants in the exercise intervention group also underwent a two-month exercise intervention. Specifically, a fitness instructor provided exercise instruction to each participant in the exercise intervention group to increase energy expenditure and achieve a target weight (a 3% reduction from their current weight). The fitness instructor provided exercise instruction once every two weeks. Each participant was instructed to keep a daily exercise log (diary) and to measure and record their daily activity using a provided activity monitor. In addition, each participant was instructed to install an application program (Asuken®) on their personal information terminal, which allows them to record their dietary habits, and to record their daily diet without consciously changing their diet. After the two-month exercise intervention, the participants in the exercise intervention group underwent a post-intervention physical examination similar to the pre-intervention examination.

[0105] 7) Selection of subjects for the exercise intervention group Of the 97 participants in the exercise intervention group who completed the two-month exercise intervention, 77 participants who were judged to have achieved a sufficient amount of exercise (more than 50% of the target exercise amount over the entire two months) and not have made any significant changes in their dietary intake were selected as a homogeneous group for analysis. Daily exercise records (diaries) were used primarily to determine exercise intake, with activity measurement results used as secondary evidence. Changes in dietary intake were determined using food records from the above application program.

[0106] 8) RNA preparation and sequencing The oil blotting film was cut to an appropriate size, and RNA was extracted using QIAzol® Lysis Reagent (Qiagen) according to the attached protocol. The extracted RNA was reverse-transcribed at 42°C for 90 minutes using the SuperScript VILO cDNA Synthesis kit (Life Technologies Japan, Inc.) to synthesize cDNA. The random primers included with the kit were used as primers for the reverse transcription reaction. A library containing DNA derived from the 20802 gene was prepared from the resulting cDNA by multiplex PCR. Multiplex PCR was performed using the Ion AmpliSeq Transcriptome Human Gene Expression Kit (Life Technologies Japan, Inc.) under the following conditions: 99°C for 2 minutes, (99°C for 15 seconds, 62°C for 16 minutes) x 20 cycles, hold at 4°C. The resulting PCR products were purified using Ampure XP (Beckman Coulter, Inc.), followed by buffer reconstitution, primer digestion, adapter ligation, purification, and amplification to prepare a library. The prepared library was loaded onto an Ion 540 chip and sequenced using an Ion S5 / XL system (Life Technologies Japan, Inc.). The gene from which each read sequence originated was determined by gene mapping using the hg19 AmpliSeq Transcriptome ERCC v1, the reference sequence for the human genome.

[0107] 9) Data preprocessing The read counts of each read obtained by sequencing the subjects' SSL-derived RNA obtained in 8) above were used as expression level data for each RNA, and the count values ​​corrected using DESeq2 (normalized count values) were used for analysis. However, reads with a read count of less than 1 were treated as missing values. In the dietary intervention group, data from 56 subjects in whom 30% or more of all detected gene species were detected were used for the following analysis. In the exercise intervention group, data from 65 subjects in whom 30% or more of all detected gene species were detected were used for the following analysis.

[0108] 10) Extraction of genes whose expression levels change with dietary and / or exercise intervention Using the data from the subjects in the dietary intervention group, which had undergone data preprocessing, we extracted genes (5987 genes) for which non-missing expression data was available for more than 90% of the subjects. Genes whose expression levels showed statistically significant changes between the RNA expression levels before and after dietary intervention (normalized count values) were extracted. The extraction of these genes was performed based on an adjusted p-value (threshold < 0.05) that controlled the false discovery rate (FDR) between the two groups: the RNA expression levels before and after dietary intervention (FDR < 0.05).

[0109] Similarly, using the data of subjects in the exercise intervention group that had undergone data preprocessing, genes whose expression levels showed statistically significant changes between the RNA expression levels (Normalized count values) before and after the exercise intervention were extracted from among the genes (6,817 genes) for which non-missing expression data was obtained for more than 90% of the subjects.

[0110] As a result, 271 differentially expressed genes were extracted in common from the data of both the dietary intervention group and the exercise intervention group, of which 228 genes showed increased expression after the weight loss intervention compared to before, and 43 genes showed decreased expression after the weight loss intervention compared to before. Furthermore, 822 differentially expressed genes were extracted only from the data of the dietary intervention group, but not from the data of the exercise intervention group, of which 401 genes showed increased expression after the dietary intervention compared to before, and 421 genes showed decreased expression after the dietary intervention compared to before. Furthermore, 298 differentially expressed genes were extracted only from the data of the exercise intervention group, but not from the data of the dietary intervention group, of which 212 genes showed increased expression after the exercise intervention compared to before, and 86 genes showed decreased expression after the exercise intervention compared to before.

[0111] Example 2: Selection of intake restriction alteration genes and consumption promotion alteration genes 11) Extraction of genes whose expression changes are specific to dietary and exercise interventions For each gene selected in 10), the ratio of RNA expression level after weight loss intervention to RNA expression level before weight loss intervention (RNA expression level after weight loss intervention / RNA expression level before weight loss intervention) was calculated as the expression level ratio (fold change: FC) from the RNA expression level data. Note that the "RNA expression level" for genes with expression changes extracted commonly from the data of the dietary intervention group and the exercise intervention group was taken as the average RNA expression level for all subjects used in the analysis of both groups. The "RNA expression level" for genes with expression changes extracted only from the data of the dietary intervention group was taken as the average RNA expression level for all subjects used in the analysis of the dietary intervention group. The "RNA expression level" for genes with expression changes extracted only from the data of the exercise intervention group was taken as the average RNA expression level for all subjects used in the analysis of the exercise intervention group.

[0112] From the genes selected in 10), genes that satisfied FC>2 or FC<1 / 2 (i.e., genes whose RNA expression levels changed more than two-fold before and after the weight-loss intervention) were extracted. As a result, no genes with expression changes were commonly extracted from the data of the dietary intervention group and the exercise intervention group. Furthermore, 57 genes with expression changes were extracted only from the data of the dietary intervention group, but not from the data of the exercise intervention group. Of these, 34 genes showed increased expression after the intervention compared to before, and 23 genes showed decreased expression after the intervention compared to before. Furthermore, 46 genes with expression changes were extracted only from the data of the exercise intervention group, but not from the data of the dietary intervention group. Of these, 41 genes showed increased expression after the intervention compared to before, and 5 genes showed decreased expression after the intervention compared to before.

[0113] 12) Selection of genes that alter intake restriction Of the genes extracted in 11), genes with expression changes extracted only from the data of the dietary intervention group were designated as intake restriction-varying genes, whose expression levels changed due to energy intake restriction behavior (dietary intervention) but not due to energy consumption-promoting behavior (exercise intervention).Of the intake restriction-varying genes, genes whose expression levels increased due to energy intake restriction behavior (dietary intervention) were designated as intake restriction-increasing genes, and genes whose expression levels decreased due to energy intake restriction behavior (dietary intervention) were designated as intake restriction-decreasing genes.

[0114] The selected genes that increase intake restriction are 34 genes: KIAA0146, FBXW4, LRP11, TCEA3, ANXA9, SPINK7, DGCR6L, CA13, THOC3, MYL9, RPIA, TPRG1, FAM83H, AGR2, DANCR, FAM108B1, RAB11FIP5, RPL13, TSNARE1, TFAP2A, EPHA2, NDUFA4L2, JMJD8, RASD2, PMM1, SNX21, NCK2, ALOX12B, RAB38, BSCL2, HRAS, CNKSR3, GLE1, and CDKN1C.

[0115] The genes that were selected to reduce intake restriction were 23 genes: CCDC82, LOC646214, TMEM14E, C5orf15, CCNL1, NRARP, LOC145474, IFIH1, RNF2, CRLF2, ITPRIPL2, LOC100303749, PRPF40A, MTSS1, CLNS1A, MEF2A, RB1, ANKRD44, ZNF292, NXF1, ARID4B, ZCRB1, and LPAR6.

[0116] 13) Selection of consumption-promoting variable genes Of the genes extracted in 11), genes with expression changes extracted only from the data of the exercise intervention group were designated as consumption-promoting variable genes, whose expression levels change with energy consumption-promoting behavior (exercise intervention) but do not change with energy intake-restricting behavior (dietary intervention).Of the consumption-promoting variable genes, genes whose expression levels increase with energy consumption-promoting behavior (exercise intervention) were designated as consumption-promoting increasing genes, and genes whose expression levels decrease with energy consumption-promoting behavior (exercise intervention) were designated as consumption-promoting decreasing genes.

[0117] The selected genes that promote consumption were SCEL, KIAA0284, BARX2, S100A2, KRTAP1-5, CAMK2N1, KRT13, PIP, AIF1L, GBAP1, KRTAP3-3, PRKCZ, EVPL, WBSCR22, ARG1, KRTAP5-10, SEPP1, PLK2, KRT6C, GSDMC, PLEKHG1, CDC42BPG, MAST4, GOLGA7B, CGA, SDR16C5, ARHGAP23, ATP6V0A4, RERE, KLC3, CGN, RAPGEF5, OSBPL1A, MAL, FMN1, CXADR, OCLN, COQ9, NIPAL4, BAIAP2L1, and SH3GL3.

[0118] The five genes that were selected for reducing consumption promotion were LOC100506888, OR11H12, LOC349196, USP17L6P, and USP17L5.

[0119] Example 3: Verification of the accuracy of determining diet behavior using selected genetic markers (1) 14) Calculation of the expression ratio (FC) For each subject in the dietary intervention group, the expression ratio (FC) of each intake restriction variable gene selected in 12) before and after the dietary intervention was calculated. Similarly, for each subject in the exercise intervention group, the expression ratio (FC) of each consumption promotion variable gene selected in 13) before and after the exercise intervention was calculated.

[0120] 15) Counting the number of subjects From the results of 14), for each gene that increases with intake restriction, the number of subjects with an expression level ratio (FC) value greater than 1 was counted, and for each gene that decreases with intake restriction, the number of subjects with an expression level ratio (FC) value less than 1 was counted. Similarly, from the results of 14), for each gene that increases with consumption promotion, the number of subjects with an expression level ratio (FC) value greater than 1 was counted, and for each gene that decreases with consumption promotion, the number of subjects with an expression level ratio (FC) value less than 1 was counted. An expression level ratio (FC) value greater than 1 means that the expression level has increased due to the weight loss intervention, and an expression level ratio (FC) value less than 1 means that the expression level has decreased due to the weight loss intervention.

[0121] 16) Verification of the accuracy of diet behavior assessment For each gene with elevated intake restriction, the percentage of subjects counted as having an expression level ratio (FC) greater than 1 among all subjects analyzed in the dietary intervention group was calculated as the detection rate (Table 1). Similarly, for each gene with reduced intake restriction, the percentage of subjects counted as having an expression level ratio (FC) less than 1 among all subjects analyzed in the dietary intervention group was calculated as the detection rate (Table 2). Similarly, for each gene with elevated consumption promotion, the percentage of subjects counted as having an expression level ratio (FC) greater than 1 among all subjects analyzed in the exercise intervention group was calculated as the detection rate (Table 3). Similarly, for each gene with reduced consumption promotion, the percentage of subjects counted as having an expression level ratio (FC) less than 1 among all subjects analyzed in the exercise intervention group was calculated as the detection rate (Table 4). The results are shown in Tables 1 to 4. Note that FC in Tables 1 to 4 is the FC of each gene calculated in 11).

[0122] [Table 1]

[0123] [Table 2]

[0124] [Table 3]

[0125] [Table 4]

[0126] In addition, the average, minimum, and maximum values ​​of the above-calculated judgment rates for all genes that increase intake restriction were calculated. The results are shown in Table 5. The results for genes that decrease intake restriction, genes that increase consumption promotion, and genes that decrease consumption promotion are also shown in Table 5.

[0127] [Table 5]

[0128] These results indicated that the selected intake restriction-increasing genes and intake restriction-decreasing genes could be used to determine whether dietary intervention had been performed, i.e., whether energy intake restriction behavior had been performed, at a rate of 60-80%. Similarly, the selected consumption promotion-increasing genes and consumption promotion-decreasing genes could be used to determine whether exercise intervention had been performed, i.e., whether energy consumption-promoting behavior had been performed, at a rate of 60-80%. These results indicated that each intake restriction variation gene could be used to determine whether energy intake restriction behavior had been performed. Furthermore, it was indicated that each consumption promotion variation gene could be used to determine whether energy consumption-promoting behavior had been performed.

[0129] <Example 4 Verification of the accuracy of determining diet behavior using selected genetic markers (2)> 17) Calculation of the expression ratio (FC) For each subject in the dietary intervention group and the exercise intervention group, the expression ratio (FC) of each intake restriction variable gene selected in 12) before and after the dietary intervention was calculated. Similarly, for each subject in the dietary intervention group and the exercise intervention group, the expression ratio (FC) of each consumption promotion variable gene selected in 13) before and after the exercise intervention was calculated.

[0130] 18) Assessment of each subject's dieting behavior Based on the results of 17), the number of genes that increase intake restriction with an FC greater than 1, the number of genes that decrease intake restriction with an FC less than 1, the number of genes that increase consumption promotion with an FC greater than 1, and the number of genes that decrease consumption promotion with an FC less than 1 were counted for each subject in the dietary intervention group and the exercise intervention group.

[0131] For each subject in the dietary intervention group and the exercise intervention group, the ratio of the total number of intake restriction-elevating genes with an FC greater than 1 to the total number of intake restriction-elevating genes with an FC less than 1 (first satisfaction level), when the total number of intake restriction-varying genes was set to 100%, was calculated. Similarly, for each subject in the dietary intervention group and the exercise intervention group, the ratio of the total number of consumption promotion-elevating genes with an FC greater than 1 to the total number of consumption promotion-elevating genes with an FC less than 1 (second satisfaction level), when the total number of consumption promotion-varying genes was set to 100%.

[0132] For each subject, the calculated ratio for the intake restriction variation gene (first satisfaction level) was compared with the calculated ratio for the consumption promotion variation gene (second satisfaction level). Subjects whose first satisfaction level was greater than the second satisfaction level were determined to be participants in energy intake restriction behavior, i.e., subjects belonging to the dietary intervention group. Similarly, subjects whose second satisfaction level was greater than the first satisfaction level were determined to be participants in energy consumption promotion behavior, i.e., subjects belonging to the exercise intervention group.

[0133] 19) Verification of the accuracy of diet behavior assessment Of the 56 subjects in the actual dietary intervention group, 48 were determined to belong to the dietary intervention group in 18), for a concordance rate of 85.7%. Similarly, of the 65 subjects in the actual exercise intervention group, 62 were determined to belong to the exercise intervention group in 18), for a concordance rate of 80.0%. These results demonstrate that the dieting behaviors predominantly engaged in by subjects can be accurately determined using both intake restriction variation genes and consumption promotion variation genes.

Claims

1. The method includes a step of determining whether a subject's dieting behavior is selected from an energy intake restriction behavior that restricts energy intake through food and an energy consumption promotion behavior that promotes energy consumption by increasing the amount of exercise, which has been performed by the subject during a period to be determined up to the time of collection of the biological sample, based on the expression level of at least one gene selected from the group consisting of intake restriction variable genes and consumption promotion variable genes or their expression products, which is derived from a biological sample of the subject whose dieting behavior is to be determined; the intake restriction fluctuating gene is at least one gene whose expression level fluctuates due to the energy intake restriction behavior and whose expression level does not fluctuate due to the energy consumption promoting behavior, The consumption-promoting variable gene is at least one gene whose expression level does not vary due to the energy intake restriction behavior, and whose expression level varies due to the energy consumption-promoting behavior. Judgment method.

2. The intake restriction fluctuating gene is an intake restriction increasing gene whose expression level increases due to the energy intake restriction behavior; and an intake restriction reducing gene whose expression level is reduced by the energy intake restriction behavior, The consumption-promoting variable gene is A consumption-promoting increasing gene whose expression level increases due to the energy consumption-promoting behavior; and a consumption-promoting reducing gene whose expression level is reduced by the energy consumption-promoting behavior, The intake restriction-increasing gene is at least one gene selected from the group consisting of 34 genes: KIAA0146, FBXW4, LRP11, TCEA3, ANXA9, SPINK7, DGCR6L, CA13, THOC3, MYL9, RPIA, TPRG1, FAM83H, AGR2, DANCR, FAM108B1, RAB11FIP5, RPL13, TSNARE1, TFAP2A, EPHA2, NDUFA4L2, JMJD8, RASD2, PMM1, SNX21, NCK2, ALOX12B, RAB38, BSCL2, HRAS, CNKSR3, GLE1, and CDKN1C; The intake restriction-lowering gene is at least one gene selected from the group consisting of 23 genes: CCDC82, LOC646214, TMEM14E, C5orf15, CCNL1, NRARP, LOC145474, IFIH1, RNF2, CRLF2, ITPRIPL2, LOC100303749, PRPF40A, MTSS1, CLNS1A, MEF2A, RB1, ANKRD44, ZNF292, NXF1, ARID4B, ZCRB1, and LPAR6; The consumption-promoting genes that increase in expression include SCEL, KIAA0284, BARX2, S100A2, KRTAP1-5, CAMK2N1, KRT13, PIP, AIF1L, GBAP1, KRTAP3-3, PRKCZ, EVPL, WBSCR22, ARG1, KRTAP5-10, SEPP1, PLK2, KRT6C, GSDMC, PLEKHG1, and CDC42. at least one gene selected from the group consisting of 41 genes: BPG, MAST4, GOLGA7B, CGA, SDR16C5, ARHGAP23, ATP6V0A4, RERE, KLC3, CGN, RAPGEF5, OSBPL1A, MAL, FMN1, CXADR, OCLN, COQ9, NIPAL4, BAIAP2L1, and SH3GL3; The consumption promotion reducing gene is at least one gene selected from the group consisting of five genes: LOC100506888, OR11H12, LOC349196, USP17L6P, and USP17L5; The determination method according to claim 1 .

3. In the determination step, it is determined whether or not the subject has performed the energy intake restriction behavior within the determination period based on the expression level of at least one gene selected from the group consisting of intake restriction fluctuating genes or its expression product. The determination method according to claim 1 or 2.

4. The determination step includes: a condition satisfaction determination step of determining whether or not an expression level of at least one gene selected from the group consisting of intake restriction fluctuating genes or its expression product derived from the biological sample of the subject, or a change index representing a change in the expression level derived from the biological sample of the subject during the determination period, satisfies a predetermined condition corresponding to the implementation of the energy intake restriction behavior; and a behavior determination step of determining that the energy intake restriction behavior was performed by the subject within the determination target period when it is determined that the predetermined condition is satisfied. The determination method according to claim 3.

5. The predetermined condition is: When the intake restriction fluctuating gene includes an intake restriction elevating gene whose expression level is increased by the energy intake restriction behavior, the expression level or the change index of the intake restriction elevating gene is equal to or greater than a reference value set for each gene, When the intake restriction fluctuating gene includes an intake restriction reducing gene whose expression level is reduced by the energy intake restriction behavior, the expression level or the change index of the intake restriction reducing gene is equal to or less than a reference value set for each gene, or is smaller than the reference value. The determination method according to claim 4.

6. In the determination step, it is determined whether or not the energy consumption promoting behavior has been performed by the subject within the determination period based on the expression level of at least one gene selected from the group consisting of the consumption promoting variable genes or its expression product. The determination method according to claim 1 or 2.

7. The determination step includes: a condition satisfaction determination step of determining whether or not an expression level of at least one gene selected from the group consisting of the consumption-promoting variable genes or its expression product derived from the biological sample of the subject, or a change index representing a change in the expression level derived from the biological sample of the subject during the determination period, satisfies a predetermined condition corresponding to the implementation of the energy consumption-promoting behavior; and a behavior determination step of determining that the energy consumption promoting behavior has been performed by the subject within the determination target period when it is determined that the predetermined condition is satisfied. The determination method according to claim 6.

8. The predetermined condition is: When the consumption promotion variable gene includes a consumption promotion increasing gene whose expression level increases due to the energy consumption promoting behavior, the expression level or the change index of the consumption promotion increasing gene is equal to or greater than a reference value set for each gene, When the consumption promotion variable gene includes a consumption promotion decreasing gene whose expression level is decreased by the energy consumption promoting behavior, the expression level or the change index of the consumption promotion decreasing gene is equal to or less than a reference value set for each gene, or is smaller than the reference value. The determination method according to claim 7.

9. In the determination step, the diet behavior performed by the subject during the determination period, selected from the energy intake restriction behavior and the energy consumption promotion behavior, is determined based on the expression level of at least one gene selected from the group consisting of the intake restriction variable genes or its expression product and the expression level of at least one gene selected from the group consisting of the consumption promotion variable genes or its expression product. The determination method according to claim 1 or 2.

10. The determination step includes: a first condition satisfaction determination step of determining whether or not an expression level of at least one gene selected from the group consisting of intake restriction fluctuating genes or its expression product derived from the biological sample of the subject, or a change index representing a change in the expression level derived from the biological sample of the subject during the determination period, satisfies a first condition corresponding to the implementation of the energy intake restriction behavior; a second condition satisfaction determination step of determining whether or not an expression level of at least one gene selected from the group consisting of the consumption-promoting variable genes or its expression product derived from the biological sample of the subject, or a change index representing a change in the expression level derived from the biological sample of the subject during the determination period, satisfies a second condition corresponding to the implementation of the energy consumption-promoting behavior; and a behavior determination step of determining the diet behavior performed by the subject during the determination period, the diet behavior being selected from the energy intake restriction behavior and the energy consumption promotion behavior, based on the determination result of the first condition satisfaction determination step and the determination result of the second condition satisfaction determination step. The determination method according to claim 9.

11. In the first condition satisfaction determination step, determining whether or not a change index representing a change in the expression levels of a plurality of genes selected from the group consisting of the intake restriction fluctuating genes or their expression products derived from the biological sample of the subject, or the expression levels derived from the biological sample of the subject, during the determination period, satisfies the first condition set for each gene; In the second condition satisfaction determination step, determining whether or not the expression levels of a plurality of genes selected from the group consisting of the consumption-promoting variable genes or their expression products, which are derived from the biological sample of the subject, or the change indexes representing changes in the expression levels derived from the biological sample of the subject during the determination period, satisfy the second condition set for each gene; In the behavior determination step, Calculating a first condition satisfaction degree indicating the number or proportion of the intake restriction variable genes that satisfy the first condition in the first condition satisfaction determination step, and a second condition satisfaction degree indicating the number or proportion of the consumption promotion variable genes that satisfy the second condition in the second condition satisfaction determination step; When the degree of satisfaction of the first condition is greater than the degree of satisfaction of the second condition, it is determined that the energy intake restriction behavior has been performed in preference to the energy consumption promotion behavior, and when the degree of satisfaction of the first condition is less than the degree of satisfaction of the second condition, it is determined that the energy consumption promotion behavior has been performed in preference to the energy intake restriction behavior. The determination method according to claim 10.

12. The expression level of the gene or its expression product is the expression level of mRNA. The determination method according to claim 1 or 2.

13. The biological sample is a biological sample collected non-invasively from the subject. The determination method according to claim 1 or 2.

14. The biological sample is lipids on the skin surface of the subject. The determination method according to claim 13.

15. A marker for determining a diet behavior performed by a subject, the marker being selected from an energy intake restriction behavior that restricts energy intake through meals and an energy consumption promotion behavior that promotes energy consumption by increasing the amount of exercise, The evaluation marker is the gene comprises at least one gene selected from the group consisting of an intake restriction variable gene whose expression level is changed by the energy intake restriction behavior but not by the energy consumption promotion behavior, and a consumption promotion variable gene whose expression level is not changed by the energy intake restriction behavior but is changed by the energy consumption promotion behavior, The intake restriction fluctuating gene is an intake restriction increasing gene whose expression level increases due to the energy intake restriction behavior; and an intake restriction reducing gene whose expression level is reduced by the energy intake restriction behavior, The consumption-promoting variable gene is A consumption-promoting increasing gene whose expression level increases due to the energy consumption-promoting behavior; and a consumption-promoting reducing gene whose expression level is reduced by the energy consumption-promoting behavior. The intake restriction-increasing gene is at least one gene selected from the group consisting of 34 genes: KIAA0146, FBXW4, LRP11, TCEA3, ANXA9, SPINK7, DGCR6L, CA13, THOC3, MYL9, RPIA, TPRG1, FAM83H, AGR2, DANCR, FAM108B1, RAB11FIP5, RPL13, TSNARE1, TFAP2A, EPHA2, NDUFA4L2, JMJD8, RASD2, PMM1, SNX21, NCK2, ALOX12B, RAB38, BSCL2, HRAS, CNKSR3, GLE1, and CDKN1C; The intake restriction-lowering gene is at least one gene selected from the group consisting of 23 genes: CCDC82, LOC646214, TMEM14E, C5orf15, CCNL1, NRARP, LOC145474, IFIH1, RNF2, CRLF2, ITPRIPL2, LOC100303749, PRPF40A, MTSS1, CLNS1A, MEF2A, RB1, ANKRD44, ZNF292, NXF1, ARID4B, ZCRB1, and LPAR6; The consumption-promoting genes that increase in expression include SCEL, KIAA0284, BARX2, S100A2, KRTAP1-5, CAMK2N1, KRT13, PIP, AIF1L, GBAP1, KRTAP3-3, PRKCZ, EVPL, WBSCR22, ARG1, KRTAP5-10, SEPP1, PLK2, KRT6C, GSDMC, PLEKHG1, and CDC42. at least one gene selected from the group consisting of 41 genes: BPG, MAST4, GOLGA7B, CGA, SDR16C5, ARHGAP23, ATP6V0A4, RERE, KLC3, CGN, RAPGEF5, OSBPL1A, MAL, FMN1, CXADR, OCLN, COQ9, NIPAL4, BAIAP2L1, and SH3GL3; The consumption promotion reducing gene is at least one gene selected from the group consisting of five genes: LOC100506888, OR11H12, LOC349196, USP17L6P, and USP17L5; Marker for evaluation.

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