CN119339932B

CN119339932B - Systems and kits for HB risk screening and early diagnosis in fetuses and infants

Info

Publication number: CN119339932B
Application number: CN202411895992.9A
Authority: CN
Inventors: 董瑞; 戴菽阳; 李仪; 陈德芊; 胡方志; 黄梦飞; 邓伟
Original assignee: Shanghai Chengran Biology Technology Co ltd
Current assignee: Childrens Hospital of Fudan University
Priority date: 2024-12-23
Filing date: 2024-12-23
Publication date: 2025-03-21
Anticipated expiration: 2044-12-23
Also published as: CN119339932A

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 hepatoblastoma in children.

Description

System and kit for HB risk screening and early diagnosis for fetuses and infants

Technical Field

The invention relates to the technical field of medical diagnosis, in particular to a system and a kit for HB risk screening and early diagnosis of fetuses and infants.

Background

Hepatoblastomas (Hepatoblastoma, HB) are a rare malignancy, almost exclusively found in young children, the most common primary malignancy in children. Hepatoblastomas are malignant embryogenic tumors with multiple differentiation patterns, accounting for about 2/3,90% of cases of childhood primary liver tumors occurring in children under 5 years of age, mainly before 2 years of age. The hepatoblastoma is asymptomatic in early stage, and the sick children often visit abdominal mass which is found accidentally. About 70% of the infants at the initial diagnosis are in clinical stage III or even more, and the time for completely killing the tumor by combining surgery and radiotherapy and chemotherapy is missed, so that the prognosis of the infants is poor. Most infants have abnormally elevated serum alpha-fetoprotein (AFP), but the marker lacks the specificity of HB diagnosis because AFP is physiologically elevated in infancy, and inflammation and benign lesions of the liver also cause the elevation of AFP. Therefore, the HB diagnostic markers with higher specificity are searched, which is helpful for early diagnosis of HB infants, thereby improving the treatment effect of the infants.

With the rapid development of gene detection technology, the liquid biopsy has increasingly prominent effect in tumor accurate medical treatment, and provides a new direction for early screening and early diagnosis of tumors. The liquid biopsy can realize early diagnosis and real-time monitoring of tumors by detecting indexes such as trace tumor-derived circulating tumor DNA (ctDNA), circulating cell free DNA (cfDNA), exosome (exosome) content and the like in body fluids such as blood, urine and the like, and has the advantages of accuracy, noninvasive property, high efficiency, high repeatability and the like.

Although the application value of cfDNA and ctDNA in tumor screening and diagnosis has been demonstrated, it is mostly applied to adults.

In view of this, the present invention has been made.

Disclosure of Invention

The object of the present invention is to provide a system and kit for HB risk screening and early diagnosis of fetuses and infants. The invention can diagnose HB risk of infants in fetal period or 24 months after birth, has higher specificity and sensibility, realizes prenatal risk prompt, and is beneficial to early clinical intervention of HB infants.

The invention is realized in the following way:

In a first aspect, the invention provides a hepatoblastoma risk screening system for a fetus, comprising an information acquisition module, a calculation module and a diagnosis module;

the information acquisition module is used for executing the step of acquiring the ready value of a target miRNA mature body and an isomer thereof in a sample to be detected, wherein the isomer comprises a first isomer and a second isomer, the first isomer lacks 1 nucleotide relative to the 3 'end of the target miRNA mature body, and the second isomer lacks 2 nucleotides relative to the 3' end of the target miRNA mature body;

the calculation module is used for executing the step of substituting the ready value into a calculation model calculation threshold value;

The diagnosis module is used for executing the step of judging the risk condition of the fetus to be screened for hepatoblastoma according to the threshold value;

the target miRNA mature body comprises 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.

The hepatoblastoma risk screening system for the fetus can identify or screen the hepatoblastoma risk of the fetus at pregnancy, has higher specificity and sensitivity, realizes prenatal risk prompt, and is beneficial to early clinical intervention on HB infants.

Optionally, in some embodiments of the present invention, 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;

Wherein R1-R15 represent the ratio 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 the ready of its respective second isomer;

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 hsa-let-7a-5p mature miRNA, and R _{-2nt -hsa-let-7a-5p} represents the Reads value of the second isoform of hsa-let-7a-5p (2 nucleotides deleted at the 3' end). The calculation of R2-R15 is similarly understood and will not be repeated here

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 ratio of the reads 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, respectively, to their respective first isomers.

For example, S1=R _{hsa-let-7a-5p}/R_{-1nt -hsa-let-7a-5p}; R_{hsa-let-7a-5p} represents the Reads value of the hsa-let-7a-5p mature miRNA, and R _{-1nt -hsa-let-7a-5p} represents the Reads value of the first isoform of hsa-let-7a-5p (1 nucleotide deleted at the 3' end). The calculations of S2-S15 are similarly understood and will not be described in detail.

Ratio1 and Ratio2 represent the risk threshold for a fetus to be screened for hepatoblastoma.

Optionally, in some embodiments of the present invention, the diagnostic module determines according to the Ratio1 and Ratio2 thresholds in the following manner:

If Ratio1+Ratio2 is more than or equal to 16.80, judging that the fetus to be screened is HB high risk, if Ratio1+Ratio2 is less than or equal to 11.68, judging that the fetus to be screened is HB low risk, if 11.68< Ratio1+Ratio2<16.80, judging that the fetus to be screened cannot be judged, and suggesting to strengthen follow-up.

Alternatively, in some embodiments of the invention, the test sample is from the plasma exosomes of the maternal host of the fetus to be screened.

Alternatively, in some embodiments of the invention, the mother is at a pregnancy of more than 30 weeks.

Alternatively, in some embodiments of the invention, the parent is at 30 weeks of gestation, 31 weeks of gestation, 32 weeks of gestation, 33 weeks of gestation, 34 weeks of gestation, 35 weeks of gestation, 36 weeks of gestation, 37 weeks of gestation, 38 weeks of gestation, or 40 weeks of gestation.

In another aspect, the present invention provides a hepatoblastoma early diagnosis system for infants, comprising an information acquisition module, a calculation module, and a diagnosis module;

The information acquisition module is used for executing the step of acquiring the ready value of a target miRNA mature body and an isomer thereof in a sample to be detected, wherein the isomer lacks 1 nucleotide relative to the 3' end of the target miRNA mature body;

the diagnosis module is used for executing the step of judging the risk condition of the infant to be diagnosed for hepatoblastoma according to the threshold value;

the target miRNA mature body comprises the following 15 miRNA molecules:

The early diagnosis system for the hepatoblastoma of the infant can perform early diagnosis of the hepatoblastoma of the infant after birth, has higher specificity and sensitivity, and is beneficial to early clinical intervention of HB infants.

Alternatively, in some embodiments of the invention, the infant to be diagnosed is an infant 24 months after birth.

The early diagnosis system can realize accurate identification of the infant suffering from hepatoblastoma within 24 months after birth, and the diagnosis AUC exceeds 0.95.

Alternatively, in some embodiments of the invention, the mathematical formula of the computational model is as follows:

S1-S15 represent the ratio of reads 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, respectively, to their respective isomers (i.e., 1 nucleotide deleted at the 3' end relative to the mature form);

Ratio2 represents the risk threshold for the infant to be diagnosed for hepatoblastoma.

Optionally, in some embodiments of the present invention, the diagnostic module determines according to the Ratio2 threshold in the following manner:

If Ratio2 is more than or equal to 10.92, judging that the infant to be diagnosed is HB high risk; if Ratio2 is less than or equal to 7.22, judging that the infant to be diagnosed is HB low risk, and if Ratio2 is 7.22< 10.92, judging that the infant cannot be judged, and suggesting to strengthen follow-up.

Alternatively, in some embodiments of the invention, the sample to be tested is from the plasma exosomes of the infant to be diagnosed.

It should be noted that sequencing reads of miRNAs and their isomers can be determined by sequencing techniques or qRCR commonly known in the art, for example, iSeq, miniSeq, miSeq, nextSeq, and NextSeq2000, novaSeq, and 6000 sequencers of illumine company based on PE150 or SE50 sequencing techniques, and MGI sequencing platform of Huada gene company based on SE100, PE100, and PE150 sequencing techniques, and the like.

The extraction and separation of the plasma small extracellular vesicles and the enrichment of miRNAs and isomers thereof can be carried out by using conventional techniques and kits in the art.

Optionally, in some embodiments of the present invention, the system further comprises a result display module for displaying the diagnosis conclusion reached by the diagnosis module.

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

In another aspect, the present invention provides a hepatoblastoma early diagnosis kit for a fetus or infant, the kit comprising a detection reagent for detecting a miRNA mature body or an isomer thereof:

the miRNA mature body comprises the following 15 miRNA molecules:

Alternatively, in some embodiments of the invention, the isomers comprise a first isomer that lacks 1 nucleotide relative to the 3 'end of the target miRNA mature form or/and a second isomer that lacks 2 nucleotides relative to the 3' end of the target miRNA mature form.

The kit provided by the invention can be used for early diagnosis of hepatoblastoma of a fetus or an infant aiming at detection of miRNA molecules or isomers thereof in the 15, and has higher specificity and sensitivity.

The nucleotide sequence of the miRNA mature body is as follows:

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

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

Fig. 1 is a schematic diagram of a detection flow of the risk screening method and the early diagnosis method provided in embodiments 1 and 2 of the present aspect.

FIG. 2 is a ROC curve of HB early diagnosis method for infant in example 4.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.

The features and capabilities of the present invention are described in further detail below in connection with the examples.

Example 1

The present embodiment provides a risk screening method for hepatoblastoma of a fetus, comprising the steps of:

(1) Obtaining ready values of mirnas and isomers thereof in exosomes:

Extracting miRNA from plasma exosomes of pregnant women, and sequencing to obtain reads values of 15 miRNA molecules and first isomer and second isomer thereof;

(2) Calculating a risk threshold value, and substituting each ready value into a Ratio1 model and a Ratio2 model to obtain a risk judgment threshold value:

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

The formula of mature hsa-let-7a-5p/-2 nt-hsa-let-7 a-5p refers to the ratio of reads of mature body miRNAhsa-let-7a-5p to reads of an isomer lacking 2 nucleotides (-2 nt) at its 3' end, other ratios are similarly understood;

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

The formula is that mature hsa-let-7a-5p/-1 nt-hsa-let-7 a-5p refers to the ratio between reads of mature miRNA hsa-let-7a-5p and reads of isomer lacking 1 nucleotide (-1 nt) at the 3' end thereof;

(3) Judging the risk of the fetus to suffer from HB based on the Ratio1 and the Ratio2, wherein if Ratio1+ratio2 is more than or equal to 16.80, the fetus to be screened is judged to be HB high risk, if Ratio1+ratio2 is less than or equal to 11.68, the fetus to be screened is judged to be HB low risk, and if Ratio1+ratio2 is less than or equal to 11.68, the fetus to be screened cannot be judged, and if Ratio1+ratio2 is less than or equal to 16.80, the follow-up is suggested.

Example 2

The present embodiment provides an early diagnosis method for hepatoblastoma of an infant, comprising the steps of:

(1) The ready value of miRNA in exosomes and its isoforms was obtained as in example 1.

(2) Calculating a risk threshold value, and substituting each ready value into the Ratio2 model to obtain a risk judgment threshold value:

(3) The infant HB risk is judged based on the Ratio2, if the Ratio2 is more than or equal to 10.92, the infant to be diagnosed is judged to be HB high risk, if the Ratio2 is less than or equal to 7.22, the infant to be diagnosed is judged to be HB low risk, and if the Ratio2 is 7.22< 10.92, the infant cannot be judged, and the follow-up visit is recommended.

Example 3

Specific application of the risk screening method provided in example 1:

At the 30 week gestation stage, 924 pregnant women's plasma exosomes derived mirnas were sequenced by NGS method and reports were generated based on the Ratio1+ Ratio2 model (risk screening method of example 1), predicting the low risk 896 cases of fetal HB, and follow-up at 1 and 2 years post-natal. The 28 high-risk cases of fetal HB are predicted, and follow-up is performed 6 months after birth for the 28 newborns, wherein 3 children have atypical symptoms such as abdominal pain, and 25 children have no HB symptoms. Sequencing of these 3 children with atypical symptoms and generation of reports based on Ratio2 model suggested high risk, and diagnosis of HB within 1 year of age, stage I clinical stage, and complete tumor elimination by surgery. 25 asymptomatic children re-sequenced plasma exosome-derived mirnas at 1 year old and generated reports based on the Ratio2 model, 16 of which suggested high risk and 9 low risk. The 9 children with low risk of detection report remained healthy without any HB symptom manifestation at the time of follow-up at 2 years of age. High risk 16 children were followed by 2 years of age, 13 had stage I HB confirmed by clinical diagnosis, the tumor did not progress, and complete elimination was achieved by surgery. The other 3 children stopped the follow-up endpoint and remained healthy without any HB symptom manifestations.

The positive predictive value of the Ratio1+ratio2 model (risk screening method of example 1) for fetal HB infant reached 57.14% and the negative predictive value within 24 months reached 100% (model was not followed up to 3 years old). The Ratio2 model (early diagnosis of example 2) had a positive predictive value of 84.21% for postnatal HB infants and a negative predictive value of 100% within 24 months (the model was not followed up to 3 years). More importantly, the HB infants with positive detection report are in the early stage of HB due to good risk management, and the prognosis after treatment is very good. Table 1 shows the prediction results and the follow-up diagnosis results, and fig. 1 shows a schematic diagram of the detection flow of this embodiment.

TABLE 1

Example 4

This example provides an application of the early diagnosis method of example 2

127 Cases of HB infants with definite diagnosis are grouped, 281 cases of liver benign tumor and 408 cases of inflammatory infants. Specific information is shown in table 2.

TABLE 2

Of these, the HB group contained 121 cases I and 6 cases II, and no hepatoblastoma metastasis occurred. The sensitivity and specificity of the method for early HB differential diagnosis were assessed by plasma exosome-derived microRNA NGS sequencing data and fluorescent PCR technology platform quantitative data on mature and isomeric forms of 15 microRNAs, respectively, using the method of example 2, and comparison of NGS method and fluorescent PCR method was performed by AUC value of ROC curve. ROC curves were generated with Ratio2 for 408 samples each (see fig. 2), where NGS method AUC was 0.999 (95% confidence interval 0.999-1.000), sensitivity 98.9%, specificity 99.2%, qPCR method AUC was 0.950 (95% confidence interval 0.930-0.970), sensitivity 85.4%, specificity 92.9%. The above results demonstrate that the early diagnostic method of example 2 has higher specificity and sensitivity.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A hepatoblastoma risk screening system for a fetus, characterized in that it comprises: the system comprises 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 the first isomer and the 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 perform 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:

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:

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 comprises: the system comprises 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:

6. The early diagnosis system for hepatoblastoma according to claim 5, characterized in that the mathematical formula of the calculation model is as follows:

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 hepatoblastoma early diagnosis system 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 early diagnosis system for hepatoblastoma 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 a fetus or infant, characterized in that the kit contains a detection reagent for detecting a mature miRNA or an isomer thereof:

The miRNA mature body includes the following 15 miRNA molecules:

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.