Systems and kits for HB risk screening and early diagnosis in fetuses and infants

Abstract

The present invention discloses a system and a kit for HB risk screening and early diagnosis of fetuses and infants, and relates to the field of medical diagnosis technology. The system of the present invention uses a variety of miRNA molecules and isomers thereof as biomarkers for early screening and diagnosis of hepatoblastoma, has high specificity and sensitivity, realizes early diagnosis of hepatoblastoma, and provides a diagnostic basis for earlier clinical intervention of childhood hepatoblastoma.

Description

Technical Field

[0001] The present invention relates to the field of medical diagnosis technology, and in particular to a system and a kit for HB risk screening and early diagnosis of fetuses and infants. Background Art

[0002] Hepatoblastoma (HB) is a rare malignant tumor that is almost only seen in young children. It is the most common primary malignant liver tumor in children. Hepatoblastoma is a malignant embryonic tumor with multiple differentiation patterns, accounting for about 2 / 3 of primary liver tumors in children. 90% of cases occur in children under 5 years old, mainly before 2 years old. Hepatoblastoma is mostly asymptomatic in the early stage, and children often seek medical treatment due to an accidental abdominal mass. At the time of initial diagnosis, about 70% of children are already in clinical stage III or even later, missing the opportunity to completely kill the tumor through surgery, radiotherapy and chemotherapy, resulting in a poor prognosis for children. Most children have abnormally elevated serum alpha-fetoprotein (AFP) levels, but because AFP is physiologically elevated in infancy, inflammation and benign lesions of the liver can also cause an increase in AFP, so this marker lacks specificity for HB diagnosis. Therefore, finding a more specific HB diagnostic marker will help early diagnosis of HB children, thereby improving the treatment effect of children.

[0003] With the rapid development of genetic testing technology, the role of liquid biopsy in precision medicine for tumors has become increasingly prominent, providing a new direction for early screening and diagnosis of tumors. Liquid biopsy can detect trace amounts of circulating tumor DNA (ctDNA), circulating cell-free DNA (cfDNA), and exosome contents in body fluids such as blood and urine, and can achieve early diagnosis and real-time monitoring of tumors. It has the advantages of accuracy, non-invasiveness, high efficiency, and high repeatability.

[0004] Although the application value of cfDNA and ctDNA in tumor screening and diagnosis has been confirmed, they are mostly used in adults.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The purpose of the present invention is to provide a system and kit for HB risk screening and early diagnosis of fetuses and infants. The present invention can perform HB risk diagnosis on infants and young children in the fetal period or within 24 months after birth, has high specificity and sensitivity, realizes prenatal risk warning, and helps to carry out clinical intervention on HB patients as early as possible.

[0007] The present invention is achieved in that:

[0008] In a first aspect, the present invention provides a hepatoblastoma risk screening system for a fetus, comprising: the system comprises an information acquisition module, a calculation module and a diagnosis module;

[0009] The information acquisition module is used to execute the step of acquiring the reads value of the target miRNA mature form and its isomers in the sample to be tested; the isomers include a first isomer and a second isomer; the first isomer lacks 1 nucleotide relative to the 3' end of the target miRNA mature form; the second isomer lacks 2 nucleotides relative to the 3' end of the target miRNA mature form;

[0010] The calculation module is used to execute the step of substituting the reads value into the calculation model to calculate the threshold value;

[0011] The diagnostic module is used to execute the step of determining the risk of hepatoblastoma in the fetus to be screened according to the threshold value;

[0012] The target miRNA mature body includes the following 15 miRNA molecules:

[0013] hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR -223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p.

[0014] The hepatoblastoma risk screening system for the fetus provided by the present invention can identify or screen the risk of hepatoblastoma for the fetus during pregnancy, has high specificity and sensitivity, realizes prenatal risk warning, and helps to carry out clinical intervention for HB children at an early stage.

[0015] Optionally, in some embodiments of the present invention, the calculation model includes a Ratio1 model and a Ratio2 model:

[0016] The mathematical formula of the Ratio1 model is as follows:

[0017] Ratio1=4.8×(R1+R2+R3)+0.6×R4+2.5×R5+0.9×R7+0.2×R7+0.7×R8+1.8×R9+3.6×R10+5.6×(R11+R12)+8.0×R13+0.3×R14+0.5×R15;

[0018] Among them, R1-R15 represent the read ratios of hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR-223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p to their respective second isoforms, respectively;

[0019] For example: R1=R hsa-let-7a-5p / R -2nt -hsa-let-7a-5p ; R hsa-let-7a-5p Represents the reads value of the mature miRNA hsa-let-7a-5p; R -2nt -hsa-let-7a-5p Represents the Reads value of the second isomer of hsa-let-7a-5p (2 nucleotides missing at the 3' end). The calculation of R2-R15 is similar and will not be repeated here.

[0020] The mathematical formula of the Ratio2 model is as follows:

[0021] Ratio2=1.8×S1+1.6×S2+5.0×S3+1.5×S4+6.4×S5+0.2×S6+0.5×S7+0.5×S8+4.5×S9+1.6×S10+1.8×(S11+S12)+4.0×S13+0.5×S14+2.5×S15;

[0022] S1-S15 represent the reads ratios of hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR-223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p to their respective first isoforms, respectively.

[0023] For example: S1=R hsa-let-7a-5p / R -1nt -hsa-let-7a-5p ; R hsa-let-7a-5pRepresents the reads value of the mature miRNA hsa-let-7a-5p; R -1nt -hsa-let-7a-5p Represents the reads value of the first isomer of hsa-let-7a-5p (1 nucleotide missing at the 3' end). The calculation of S2-S15 is similar and will not be repeated here.

[0024] Ratio1 and Ratio2 represent the risk thresholds for hepatoblastoma in the fetus to be screened.

[0025] Optionally, in some embodiments of the present invention, the diagnosis module makes a judgment based on the Ratio1 and Ratio2 thresholds in the following manner:

[0026] If Ratio1+Ratio2≥16.80, the fetus to be screened is judged to be at high risk of HB; if Ratio1+Ratio2≤11.68, the fetus to be screened is judged to be at low risk of HB; if 11.68<Ratio1+Ratio2<16.80, it cannot be judged and it is recommended to strengthen follow-up.

[0027] Optionally, in some embodiments of the present invention, the sample to be tested is derived from plasma exosomes of the mother of the fetus to be screened.

[0028] Optionally, in some embodiments of the present invention, the mother is over 30 weeks of gestation.

[0029] Optionally, in some embodiments of the present invention, the mother is at week 30, week 31, week 32, week 33, week 34, week 35, week 36, week 37, week 38, week 38 or week 40 of pregnancy.

[0030] On the other hand, the present invention provides an early diagnosis system for hepatoblastoma in infants, comprising: the system comprises an information acquisition module, a calculation module and a diagnosis module;

[0031] The information acquisition module is used to execute the step of acquiring the reads value of the target miRNA mature form and its isomer in the sample to be tested; the isomer lacks 1 nucleotide relative to the 3' end of the target miRNA mature form;

[0032] The calculation module is used to execute the step of substituting the reads value into the calculation model to calculate the threshold value;

[0033] The diagnostic module is used to execute the step of determining the risk of hepatoblastoma of the infant to be diagnosed according to the threshold value;

[0034] The target miRNA mature body includes the following 15 miRNA molecules:

[0035] hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR -223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p.

[0036] The early diagnosis system for hepatoblastoma in infants provided by the present invention can perform early diagnosis of hepatoblastoma in infants after birth, has high specificity and sensitivity, and is helpful for early clinical intervention of infants with HB.

[0037] Optionally, in some embodiments of the present invention, the infant to be diagnosed is an infant 24 months after birth.

[0038] The above-mentioned early diagnosis system can accurately identify infants with hepatoblastoma within 24 months after birth, with a diagnostic AUC exceeding 0.95.

[0039] Optionally, in some embodiments of the present invention, the mathematical formula of the calculation model is as follows:

[0040] Ratio2=1.8×S1+1.6×S2+5.0×S3+1.5×S4+6.4×S5+0.2×S6+0.5×S7+0.5×S8+4.5×S9+1.6×S10+1.8×(S11+S12)+4.0×S13+0.5×S14+2.5×S15;

[0041] S1-S15 represent the read ratios of hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR-223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p to their respective isoforms (i.e., one nucleotide is missing at the 3' end relative to the mature form), respectively;

[0042] Ratio2 represents the risk threshold for hepatoblastoma in infants to be diagnosed.

[0043] Optionally, in some embodiments of the present invention, the diagnosis module makes a judgment according to the Ratio2 threshold value in the following manner:

[0044] If Ratio2≥10.92, the infant to be diagnosed is judged to be at high risk of HB; if Ratio2≤7.22, the infant to be diagnosed is judged to be at low risk of HB; if 7.22<Ratio2<10.92, it cannot be judged and it is recommended to strengthen follow-up.

[0045] Optionally, in some embodiments of the present invention, the sample to be tested is derived from plasma exosomes of the infant to be diagnosed.

[0046] It should be noted that the sequencing reads of miRNA and its isoforms can be measured by common sequencing technologies or qRCR in this field, such as Illumina's iSeq100, MiniSeq, MiSeq, NextSeq550, NextSeq2000, and NovaSeq6000 sequencers based on PE150 or SE50 sequencing technologies; BGI's MGI sequencing platform is based on SE100, PE100, and PE150 sequencing technologies.

[0047] It should be noted that the methods for extracting and separating plasma small extracellular vesicles and enriching miRNA and its isoforms used in the present invention can be carried out using conventional techniques and kits in the art.

[0048] Optionally, in some embodiments of the present invention, the system further comprises a result display module, and the result display module is used to display the diagnosis conclusion obtained by the diagnosis module.

[0049] Optionally, in some embodiments of the present invention, the result display module displays the diagnosis result by screen display, voice broadcast or printing.

[0050] In another aspect, the present invention provides a kit for early diagnosis of hepatoblastoma in a fetus or infant, the kit comprising a detection reagent for detecting a mature miRNA or an isoform thereof:

[0051] The miRNA mature body includes the following 15 miRNA molecules:

[0052] hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR -223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p.

[0053] Optionally, in some embodiments of the present invention, the isomers include a first isomer and / or a second isomer; the first isomer lacks 1 nucleotide relative to the 3' end of the target miRNA mature body; the second isomer lacks 2 nucleotides relative to the 3' end of the target miRNA mature body.

[0054] The kit provided by the present invention can detect the above 15 miRNA molecules or their isomers and is used for early diagnosis of hepatoblastoma in fetuses or infants, with high specificity and sensitivity.

[0055] The nucleotide sequence of the above miRNA mature form is as follows:

[0056] miRNA Name Nucleotide Sequence: SEQ ID NO: hsa-let-7a-5p 5’-UGAGGUAGUAGGUUGUAUAGUU-3’ 1 hsa-let-7b-5p 5’-UGAGGUAGUAGGUUGUGUGGUU-3’ 2 hsa-let-7f-5p 5’-UGAGGUAGUAGAUUGUAUAGUU-3’ 3 hsa-miR-122-5p 5’-UGGAGUGUGACAAUGGUGUUUG-3’ 4 hsa-miR-148a-3p 5’-UCAGUGCACUACAGAACUUUGU-3’ 5 hsa-miR-16-5p 5’-UAGCAGCACGUAAAUAUUGGCG-3’ 6 hsa-miR-191-5p 5’-CAACGGAAUCCCAAAAGCAGCUG-3’ 7 hsa-miR-223-3p 5’-UGUCAGUUUGUCAAAUACCCCA-3’ 8 hsa-miR-26a-5p 5’-UUCAAGUAAUCCAGGAUAGGCU-3’ 9 hsa-miR-27b-3p 5’-UUCACAGUGGCUAAGUUCUGC-3’ 10 hsa-miR-30d-5p 5’-UGUAAACAUCCCCGACUGGAAG-3’ 11 hsa-miR-30e-5p 5’-UGUAAACAUCCUUGACUGGAAG-3’ 12 hsa-miR-423-5p 5’-UGAGGGGCAGAGAGCGAGACUUU-3’ 13 hsa-miR-93-3p 5’-ACUGCUGAGCUAGCACUUCCCG-3’ 14 hsa-miR-9-3p 5’-AUAAAGCUAGAUAACCGAAAGU-3’ 15 BRIEF DESCRIPTION OF THE DRAWINGS

[0057] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0058] Figure 1 Schematic diagram of the detection process of the risk screening method and early diagnosis method provided in Example 1 and Example 2 of this aspect.

[0059] Figure 2 This is the ROC curve of the early diagnosis method for HB in infants in Example 4. DETAILED DESCRIPTION

[0060] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0061] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0062] Example 1

[0063] This embodiment provides a risk screening method for fetal hepatoblastoma, comprising the following steps:

[0064] (1) Obtain the reads value of miRNA and its isoforms in exosomes:

[0065] The miRNA derived from plasma exosomes of pregnant women was extracted and sequenced to obtain the read values ​​of 15 miRNA molecules and their first isomers and second isomers;

[0066] (2) Calculate the risk threshold and substitute each read value into the Ratio1 model and the Ratio2 model to obtain the risk judgment threshold:

[0067] Ratio1 = 4.8 × (mature hsa-let-7a-5p / -2nt -hsa-let-7a-5p+mature hsa-let-7b-5p / -2nt-hsa-let-7b-5p+mature hsa-let-7f-5p / -2nt-h sa-let-7f-5p)+0.6×(mature hsa-miR-122-5p / -2nt-hsa-miR-122-5p)+2.5×(mature hsa-miR-1 48a-3p / -2nt-hsa-miR-148a-3p)+0.9×(mature hsa-miR-16-5p / -2nt-hsa-miR-16-5p)+0.2 ×(mature hsa-miR-191-5p / -2nt-hsa-miR-191-5p)+0.7×(mature hsa-miR-223-3p / -2nt-hsa-mi hsa-miR-223-3p)+1.8×(mature hsa-miR-26a-5p / -2nt-hsa-miR-26a-5p)+3.6×(mature hsa-miR-27b-3p / -2nt-hsa-miR-27b-3p)+5.6×(mature hsa-miR-30d-5p / -2nt-miR-30d-5p+mature hsa-miR-3 0e-5p / -2nt-miR-30e-5p)+8.0×(mature hsa-miR-423-5p / -2nt-hsa-miR-423-5p)+0.3×(mature hsa-miR-93-3p / -2nt-hsa-miR-93-3p)+0.5×(mature hsa-miR-9-3p / -2nt-hsa-miR-9-3p);

[0068] In the formula: mature hsa-let-7a-5p / -2nt -hsa-let-7a-5p refers to the ratio between the reads of mature miRNA hsa-let-7a-5p and the reads of its isomer with 2 nucleotides missing (-2nt) at the 3' end; other ratios are understood similarly;

[0069] Ratio2 = 1.8 × (mature hsa-let-7a-5p / -1nt-hsa-let-7a-5p) + 1.6 × (mature hsa-let-7b-5p / -1nt-hsa-let-7b-5p) + 5.0 × (mature hsa-let-7f-5p / -1nt-hsa-let-7f-5p) + 1.5 × (mature hsa-miR-122-5p / -1nt -hsa-miR-122-5p) + 6.4×(mature hsa-miR-148a-3p / -1nt-hsa-miR-148a-3p) + 0.2×(mature hsa-miR-16-5p / -1nt-hsa-miR-16-5p) + 0.5×(mature hsa-miR-191-5p / -1nt-hsa-miR-191-5p) + 0.5×(mature hsa -miR-223-3p / -1nt-hsa-miR-223-3p)+4.5×(mature hsa-miR-26a-5p / -1nt-hsa-miR-26a-5p)+1.6×(mature hsa-miR-27b-3p / -1nt-hsa-miR-27b-3p)+1.8×(mature hsa-miR-30d-5p / -1nt-miR-30d-5p +mature hsa-miR-30e-5p / -1nt-miR-30e-5p)+4.0×(mature hsa-miR-423-5p / -1nt-hsa-miR-423-5p)+0.5×(mature hsa-miR-93-3p / -1nt-hsa-miR-93-3p)+2.5×(mature hsa-miR-9-3p / -1nt-hsa-miR-9-3p)

[0070] In the formula: mature hsa-let-7a-5p / -1nt -hsa-let-7a-5p refers to the ratio between the reads of mature miRNA hsa-let-7a-5p and the reads of its isomer with one nucleotide missing (-1nt) at the 3' end; the read ratios of other molecules are understood similarly;

[0071] (3) The risk of fetal HB is judged based on Ratio1 and Ratio2: if Ratio1+Ratio2≥16.80, the fetus to be screened is judged to be at high risk of HB; if Ratio1+Ratio2≤11.68, the fetus to be screened is judged to be at low risk of HB; if 11.68<Ratio1+Ratio2<16.80, it cannot be judged and it is recommended to strengthen follow-up.

[0072] Example 2

[0073] This embodiment provides an early diagnosis method for hepatoblastoma in infants, comprising the following steps:

[0074] (1) Obtaining the reads value of miRNA and its isoforms in exosomes: the same as Example 1.

[0075] (2) Calculate the risk threshold and substitute each read value into the above Ratio2 model to obtain the risk judgment threshold:

[0076] (3) The risk of HB in infants should be judged based on Ratio2: if Ratio2 ≥ 10.92, the infant is judged to be at high risk of HB; if Ratio2 ≤ 7.22, the infant is judged to be at low risk of HB; if 7.22 < Ratio2 < 10.92, no judgment can be made and follow-up is recommended.

[0077] Example 3

[0078] Specific application of the risk screening method provided in Example 1:

[0079] At 30 weeks of pregnancy, the miRNA derived from plasma exosomes of 924 pregnant women was sequenced by NGS method and a report was generated based on the Ratio1+Ratio2 model (risk screening method of Example 1), predicting 896 cases of low risk of fetal HB, and followed up one and two years after the birth of the newborn. 28 cases of high risk of fetal HB were predicted, and these 28 newborns were followed up 6 months after birth. Among them, 3 children had atypical symptoms such as abdominal pain, and 25 children had no symptoms of HB. The plasma exosome-derived miRNA of these 3 children with atypical symptoms was sequenced and the report generated based on the Ratio2 model indicated high risk, and they were diagnosed with HB within 1 year old, with clinical stage I, and the tumor was completely eliminated by surgery. 25 asymptomatic children were again sequenced for plasma exosome-derived miRNA at the age of 1 year old and a report was generated based on the Ratio2 model, of which 16 cases indicated high risk and 9 cases were low risk. The 9 children with low risk in the test report remained healthy and had no symptoms of HB when they were followed up at the age of 2 years old. The 16 high-risk children were followed up at the age of 2 years. Thirteen of them were clinically diagnosed with stage I HB, and their tumors did not progress and were completely eliminated through surgery. The other three children remained healthy and had no HB symptoms at the end of the follow-up.

[0080] The positive predictive value of the Ratio1+Ratio2 model (the risk screening method of Example 1) for infants with HB in the fetal period reached 57.14%, and the negative predictive value within 24 months reached 100% (the model was not followed up continuously until after 3 years of age). The positive predictive value of the Ratio2 model (early diagnosis of Example 2) for infants with HB after birth reached 84.21%, and the negative predictive value within 24 months reached 100% (the model was not followed up continuously until after 3 years of age). More importantly, the HB children with positive test reports were in the early stages of HB due to good risk management, and their prognosis after treatment was very good. Table 1 shows the prediction results and follow-up diagnosis results. Figure 1 A schematic diagram showing the detection process of this embodiment.

[0081] Table 1

[0082]

[0083] Example 4

[0084] This embodiment provides an application of the early diagnosis method of embodiment 2

[0085] A total of 408 children were enrolled, including 127 confirmed HB patients and 281 children with benign liver tumors and inflammation. Detailed information is shown in Table 2.

[0086] Table 2

[0087]

[0088] Among them, the HB group included 121 cases of stage I and 6 cases of stage II, and none of them had hepatoblastoma metastasis. The sensitivity and specificity of this method in the differential diagnosis of early HB were evaluated using the method of Example 2 by using the NGS sequencing data of microRNA derived from plasma exosomes and the quantitative data of mature and isoforms of 15 microRNAs using the fluorescent PCR technology platform, and the AUC value of the ROC curve was used to compare the NGS method and the fluorescent PCR method. The ROC curve was generated using the Ratio2 of each of the 408 samples (see Figure 2 ), among which the AUC of NGS method was 0.999 (95% confidence interval 0.999-1.000), sensitivity 98.9%, and specificity 99.2%; the AUC of qPCR method was 0.950 (95% confidence interval 0.930-0.970), sensitivity 85.4%, and specificity 92.9%. The above results show that the early diagnosis method of Example 2 has high specificity and sensitivity.

[0089] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A hepatoblastoma risk screening system for a fetus, characterized in that: It includes: The system includes an information acquisition module, a calculation module and a diagnosis module; The information acquisition module is used to execute the step of acquiring the reads value of the target miRNA mature form and its isomers in the sample to be tested; the isomers include a first isomer and a second isomer; the first isomer lacks 1 nucleotide relative to the 3' end of the target miRNA mature form; the second isomer lacks 2 nucleotides relative to the 3' end of the target miRNA mature form; The calculation module is used to execute the step of substituting the reads value into the calculation model to calculate the threshold value; The diagnostic module is used to execute the step of determining the risk of hepatoblastoma in the fetus to be screened according to the threshold value; The target miRNA mature body includes the following 15 miRNA molecules: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR -223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p.

2. The hepatoblastoma risk screening system according to claim 1, characterized in that: The calculation model includes a Ratio1 model and a Ratio2 model: The mathematical formula of the Ratio1 model is as follows: Ratio1=4.8×(R1+R2+R3)+0.6×R4+2.5×R5+0.9×R7+0.2×R7+0.7×R8+1.8×R9+3.6×R10+5.6×(R11+R12)+8.0×R13+0.3×R14+0.5×R15; Among them, R1-R15 represent the read ratios of hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR-223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p to their respective second isoforms, respectively; The mathematical formula of the Ratio2 model is as follows: Ratio2=1.8×S1+1.6×S2+5.0×S3+1.5×S4+6.4×S5+0.2×S6+0.5×S7+0.5×S8+4.5×S9+1.6×S10+1.8×(S11+S12)+4.0×S13+0.5×S14+2.5×S15; S1-S15 represent the read ratios of hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR-223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p to their respective first isoforms, respectively; Ratio1 and Ratio2 represent the risk thresholds for hepatoblastoma in the fetus to be screened.

3. The hepatoblastoma risk screening system according to claim 2, characterized in that: The diagnosis module makes a judgment based on the Ratio1 and Ratio2 thresholds in the following manner: If Ratio1+Ratio2≥16.80, the fetus to be screened is judged to be at high risk of HB; if Ratio1+Ratio2≤11.68, the fetus to be screened is judged to be at low risk of HB; if 11.68<Ratio1+Ratio2<16.80, it cannot be judged and it is recommended to strengthen follow-up.

4. The hepatoblastoma risk screening system according to any one of claims 1 to 3, characterized in that: The sample to be tested is derived from plasma exosomes of the mother of the fetus to be screened.

5. An early diagnosis system for hepatoblastoma in infants, characterized in that: It includes: The system includes an information acquisition module, a calculation module and a diagnosis module; The information acquisition module is used to execute the step of acquiring the reads value of the target miRNA mature form and its isomer in the sample to be tested; the isomer lacks 1 nucleotide relative to the 3' end of the target miRNA mature form; The calculation module is used to execute the step of substituting the reads value into the calculation model to calculate the threshold value; The diagnostic module is used to execute the step of determining the risk of hepatoblastoma of the infant to be diagnosed according to the threshold value; The target miRNA mature body includes the following 15 miRNA molecules: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR -223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p.

6. The early diagnosis system for hepatoblastoma according to claim 5, characterized in that: The mathematical formula of the calculation model is as follows: Ratio2=1.8×S1+1.6×S2+5.0×S3+1.5×S4+6.4×S5+0.2×S6+0.5×S7+0.5×S8+4.5×S9+1.6×S10+1.8×(S11+S12)+4.0×S13+0.5×S14+2.5×S15; S1-S15 represent the read ratios of hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR-223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p to their respective isoforms, respectively; Ratio2 represents the risk threshold for hepatoblastoma in infants to be diagnosed.

7. The early diagnosis system for hepatoblastoma according to claim 5, characterized in that: The diagnosis module makes a judgment based on the Ratio2 threshold in the following manner: If Ratio2≥10.92, the infant to be diagnosed is judged to be at high risk of HB; if Ratio2≤7.22, the infant to be diagnosed is judged to be at low risk of HB; if 7.22<Ratio2<10.92, it cannot be judged and it is recommended to strengthen follow-up.

8. The hepatoblastoma early diagnosis system according to any one of claims 5 to 7, characterized in that: The sample to be tested is derived from plasma exosomes of the infant to be diagnosed.

9. A kit for early diagnosis of hepatoblastoma in fetuses or infants, characterized in that: The kit contains a detection reagent for detecting a mature miRNA or its isoforms: The miRNA mature body includes the following 15 miRNA molecules: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7f-5p, hsa-miR-122-5p, hsa-miR-148a-3p, hsa-miR-16-5p, hsa-miR-191-5p, hsa-miR -223-3p, hsa-miR-26a-5p, hsa-miR-27b-3p, hsa-miR-30d-5p, hsa-miR-30e-5p, hsa-miR-423-5p, hsa-miR-93-3p and hsa-miR-9-3p.

10. The kit according to claim 9, characterized in that The isomers include a first isomer and / or a second isomer; the first isomer lacks 1 nucleotide relative to the 3' end of the miRNA mature body; the second isomer lacks 2 nucleotides relative to the 3' end of the miRNA mature body.

Images