CN109979596B - Method for establishing prediction risk model of maternal serum marker for fetal hypospadias - Google Patents
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Abstract
The invention discloses a method for establishing a prediction risk model of maternal serum AFP and free beta-HCG on fetal hypospadias, which comprises the following steps: (1) dividing the pregnant women into a case group and a control group according to the existence of the fetal hypospadias; (2) detecting the AFP and free beta-HCG levels of the serum of 2 groups of pregnant women by adopting a time-resolved immunofluorescence method; (3) and during detection, detecting the AFP and free beta-HCG levels of the serum of the pregnant woman to be detected by adopting a time-resolved immunofluorescence method, and screening by using a risk calculation model constructed by the AFP and free beta-HCG. The invention has the beneficial effects that: the method has high sensitivity and specificity when maternal serum AFP and free beta-HCG are used for jointly screening hypospadias in the middle pregnancy period, and can become a new marker for predicting the hypospadias of fetuses; a risk calculation model constructed by AFP and free beta-HCG levels of serum has great clinical value in screening the fetal hypospadias.
Description
Technical Field
The invention relates to the field of medical detection, and mainly relates to a method for establishing a prediction risk model of a maternal serum marker for fetal hypospadias.
Background
The common congenital malformation of male external genitalia characterized by abnormal opening of the urethra in the penis, ventral scrotum or perineum[1]It is one of the common malformations of human genitourinary system. Although many documents report that the cause of hypospadias has been identified, it is caused by a single gene such as WT1, SF1, BMP4, etc[2]GSTT1HOXA4, IRX5, IRX6 and EYA1 genes[3]And maternal genetic polymorphisms at risk for CYP1A1, GSTM1 and GSTT1 and hypospadias[4]A mixture of multiple factors, both genetic and environmental factors such as endocrine disrupting chemicals[5-7]Smoking, low birth weight, maternal hypertension and preeclampsia, etc., most of the risk factors remain unknown[3]. The incidence rate of the current literature report is 1/300-1/125 between birth male infants[8-10]Neonatal male infant prevalence in Asia, Europe, and North America ranges from 0.3% to 0.8%[11]The incidence of inferior Malihard urethrolysis in the northern east City of Iran was 0.4%[12]The incidence of disease is on the rise worldwide. The clinical diagnosis of the hypospadias is mainly made by observing whether the abnormal opening of the urethra exists or not by the clinician with naked eyes after the baby is born. WhilePrediction of fetal hypospadias using maternal serum alpha-fetoprotein (AFP) and free human chorionic gonadotropin beta subunit (free beta-HCG) has also been rarely reported. At present, the combined examination of maternal serum AFP and free beta-HCG is widely applied to the fields of screening fetal Down syndrome, open neural tube defects and the like, so a case contrast retrospective study method is adopted in the study, on the basis of the dual screening of the Down syndrome, healthy fetal pregnant women and pregnant women which are diagnosed as hypospadias before delivery are selected as study objects, and the correlation and the diagnosis value of the AFP and free beta-HCG levels and the hypospadias are explored.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a method for establishing a prediction risk model of a maternal serum marker for fetal hypospadia.
The object of the present invention is achieved by the following technical means. A method for establishing a prediction risk model of a maternal serum marker for fetal hypospadias comprises the following steps:
(1) dividing the pregnant women into a case group and a control group according to the existence of the hypospadias of the fetus, wherein the case group is the pregnant women who are clinically confirmed to give birth to the hypospadias of the fetus, and the control group is the pregnant women who randomly draw the fetus in the same period and have normal development;
(2) detecting the AFP and free beta-HCG levels of the 2 groups of pregnant woman serum by adopting a time-resolved immunofluorescence method, constructing a risk calculation model by using the AFP and free beta-HCG, and determining an optimal cutoff value and an area AUC under a curve according to an ROC curve;
(3) and during detection, detecting the AFP and free beta-HCG levels of the serum of the pregnant woman to be detected by adopting a time-resolved immunofluorescence method, screening by using a risk calculation model constructed by the AFP and free beta-HCG, and judging that the fetus of the pregnant woman to be detected is a hypospadias fetus when the AFP and free beta-HCG levels of the serum of the pregnant woman to be detected exceed a set threshold value.
Further, the risk calculation model is: the Lifecycle-like risk value is calculated by multiplying the age risk value by the corresponding normal distribution likelihood ratio value,
the age equation: riskage=0.000627+exp-16.2395+0.286*(age-0.5)
Wherein riskageIs age risk value, age is pre-delivery age
two-dimensional normal distribution likelihood calculation formula:
wherein sigma is the standard deviation of the corresponding index, rho is the correlation coefficient of the two indexes, mu is the mean value of the sample, where x denotes the logarithm of AFP MoM value, y is the logarithm of free-beta HCGMoM value, and the final hypospadias risk value:
the invention has the beneficial effects that: the method has high sensitivity and specificity when maternal serum AFP and free beta-HCG are used for jointly screening hypospadias in the middle pregnancy period, and can become a new marker for predicting the hypospadias of fetuses; a risk calculation model constructed by AFP and free beta-HCG levels of serum has great clinical value in screening the fetal hypospadias.
Drawings
FIG. 1 is a ROC curve for 3 protocols to diagnose hypospadias.
Detailed Description
The invention will be described in detail below with reference to the following drawings:
example (b): a method for establishing a prediction risk model of a maternal serum marker for fetal hypospadias comprises the following steps:
(1) dividing the pregnant women into a case group and a control group according to the existence of the hypospadias of the fetus, wherein the case group is the pregnant women who are clinically confirmed to give birth to the hypospadias of the fetus, and the control group is the pregnant women who randomly draw the fetus in the same period and have normal development;
(2) detecting the AFP and free beta-HCG levels of the 2 groups of pregnant woman serum by adopting a time-resolved immunofluorescence method, constructing a risk calculation model by using the AFP and free beta-HCG, and determining an optimal cutoff value and an area AUC under a curve according to an ROC curve;
(3) and during detection, detecting the AFP and free beta-HCG levels of the serum of the pregnant woman to be detected by adopting a time-resolved immunofluorescence method, screening by using a risk calculation model constructed by the AFP and free beta-HCG, and judging that the fetus of the pregnant woman to be detected is a hypospadias fetus when the AFP and free beta-HCG levels of the serum of the pregnant woman to be detected exceed a set threshold value.
The risk calculation model is: the Lifecycle-like risk value is calculated by multiplying the age risk value by the corresponding normal distribution likelihood ratio value,
the age equation: riskage=0.000627+exp-16.2395+0.286*(age-0.5)
Wherein riskageIs age risk value, age is pre-delivery age
two-dimensional normal distribution likelihood calculation formula:
wherein sigma is the standard deviation of the corresponding index, rho is the correlation coefficient of the two indexes, mu is the mean value of the sample, where x denotes the logarithm of AFP MoM value, y is the logarithm of free-beta HCGMoM value, and the final hypospadias risk value:
1 object and method
1.1 the subjects used the case contrast study method, and analyzed retrospectively 501628 outpatients before delivery and 15-20 pregnancies of 5 prenatal screening centers+6Single tyre of weekAmong pregnant women, 141338 were from prenatal screening centers of maternal and child healthcare institutions in the areas of late Hangzhou in Hangzhou city from 10 months 2008 to 9 months 2018, and 360290 were from 4 prenatal screening centers of maternal and child healthcare institutions in Hangzhou city from 10 months 2015 to 9 months 2018, gynecologic hospitals affiliated to Zhejiang university medical colleges, Zhejiang Xiaoshan hospital, and Hangzhou yang-rich area in Hangzhou city. Case group is hypospadias foetus diagnosed by live birth and visual observation[1]69 pregnant women (including 55 cases of simple hypospadias, hypospadias and other deformities 14 cases) and 62 pregnant women who were born at the same period of follow-up and had developed normally were randomly selected as a healthy control group. The age, pregnancy and the difference of body mass of the pregnant women in the case group and the control group have no statistical significance (P is more than 0.05), and are shown in Table 1. The study was approved by the ethical committee of hospitals and the approval number was [2018 ]]Doctor Lun examination No. (004) 01.
1.2 diagnosis and elimination of Standard cases diagnosis based on the diagnosis standards made by the China birth defects monitoring network[13]Carrying out the following steps: hypospadias is a congenital abnormality characterized by abnormal opening of the urethra to the penis, ventral scrotum or perineum. The clinical classification can be divided into: firstly, the penis is shaped; the penis is in shape; ③ scrotum type of penis; (iv) a thread-type thread. After birth, the diagnosis can be made by visual observation[1,14]. And (3) excluding: the urinary tract superior fissure, the penis bending or the prepuce stem overlong with the normal urethral opening and the false amphoteric deformity should be eliminated; double-and multiple-gestation; in combination with other medical conditions such as insulin-dependent diabetes mellitus and severe pregnancy complications; smoking; a test-tube infant; follow-up results are Down's syndrome, 18-trisomy syndrome, neural tube defects and other birth defects; the data information is not complete.
1.3 samples are collected in hospitals at all fixed points to extract 3-5 mL of fasting venous blood, serum samples are separated after 30min, 1.0-2 mL of fasting venous blood is absorbed by a disposable suction tube and placed in a special centrifugal tube, the centrifugal tube is stored in a refrigerator at the temperature of 2-8 ℃, cold chain transportation is adopted for sample transportation at low temperature, and the samples are sent for inspection within 1 week. And storing redundant serum samples in a refrigerator at the temperature of 80 ℃ below zero after the prenatal screening and detection of serology.
1.4 time-resolved fluoroimmunoassay for AFP and free β -HCG serum AFP and free β -HCG concentrations were measured using a double-labeled kit (AFP/free β -HCG) and a 1235AutoDELFIA automated time-resolved fluoroimmunoassay analyzer from PerkinElmer, USA, the levels of which were expressed in terms of median multiples (MOM), and the procedures were as described in the specification.
1.5 pregnancy outcome follow-up visit each screening object carries out third-level network follow-up visit after the birth of the newborn, and carries out routine physical examination on the newborn. According to the Chinese birth defect monitoring scheme[13]Fill in "registration card for birth defects".
1.6 establishing different models to compare the screening efficiency of the hypospadias by using a risk calculation model scheme (lifecycle-like risk value truncation scheme) constructed by AFP and free beta-HCG compared with the original single truncation scheme which independently uses AFPMOM and free beta-HCGMOM values, the MOM values of AFP and free beta-HCG obey multivariate normal distribution f (AFPMOM and free beta-HCGMOM), and the modeling method according to the risk calculation model[16]Corresponding parameters of the distribution of each index can be calculated, and the model is used as the risk of the hypospadias by calculating the distribution likelihood. Three models were constructed separately using the same principles: model one: the AFPMOM value is simply connected; model two: the free beta-HCGMOM value is simply connected; and (3) model III: AFP and free beta-HCG are combined.
1.7 statistical methods a database was built using Excel 2007 software and IBM SPSS 21.0 software was statistically processed. Data were tested for normal distribution using One-sample Kolmogorov-Smirnov. Data of normal distribution of age, weight and gestational ageIndicates that AFP and free beta-HCG results are skewed, with median and percentile [ M (P)2.5,P97.5)]And (4) showing. The baseline comparison between case and control groups was performed using two independent samples, t-test, AFP and free β -HCG levels, and rank-sum test. Plotting Receiver Operating Characteristic (ROC) curves[17]And calculating the optimal cut-off values, area under the curve (AUC), john's index for AFP and Free β hCG. P < 0.05 is statistically significant.
2 results
The AFP level of the 2.12 groups of serum AFP and free beta-HCG comparison case groups is 1.14 (0.65-3.40) MOM, which is higher than 0.96 (0.55-1.94) MOM of the control group, and the difference is statistically significant (Z is 2.831, P is 0.005). The free beta-HCG level of the pregnant women in the case group is 1.30 (0.33-19.41) MOM, which is higher than 0.84 (0.37-3.48) MOM of the control group, and the difference has statistical significance (Z is 3.131, P is 0.004). See table 2.
2.2 ROC Curve analysis of AFP or free β -HCG alone predicted hypospadias fetuses As shown in Table 3, AFP predicted the AUC of hypospadias fetuses to be 0.644 (95% CI: 0.550-0.737, P ═ 0.005). free β -HCG predicted AUC for hypospadias to be 0.659 (95% CI: 0.565-0.752, P ═ 0.002). Calculating the optimal cut-off values of AFP and free beta-HCG for screening the hypospadias according to the ROC curve respectively as follows: 0.945MOM and 1.275MOM, wherein the sensitivity, specificity and Johnson index are respectively as follows: 0.739, 0.484, 0.223; 0.522, 0.790 and 0.312.
2.3 value of AFP and free beta-HCG in combination for predicting fetal hypospadias
Method for calculating Lifecycle-like risk value[16]
Equation of age[15]:riskage=0.000627+exp-16.2395+0.286*(age-0.5)
Wherein riskageIs age risk value, age is pre-delivery age
two-dimensional normal distribution likelihood calculation formula:
wherein, σ is the standard deviation of the corresponding index, ρ is the correlation coefficient of the two indexes, and μ is the sample mean. Where x denotes the logarithm of the AFPMoM value and y is the logarithm of the free- β HCGMoM value.
Final hypospadias risk value:
the AUC of the hypospadias is predicted to be 0.700 (95% CI: 0.610-0.789, P <0.001) by combining AFP + free beta-HCG, the sensitivity is 0.551, and the specificity is 0.855. See table 3 and figure 1.
2.4 different screening protocols comparison lifecycle-like risk value cut-off protocol with single cut-off protocol for AFPMoM, free β -HCGMoM values see FIG. 1. The AUC (0.700) of the fetus with the AFP + free beta-HCG combined prediction of the hypospadias is higher than the diagnostic value of the fetus with the AFP or free beta-HCG alone prediction of the hypospadias.
Discussion of 3
Hypospadias refers to the common congenital deformity of male external genitalia characterized by abnormal opening of the urethra in the penis, abdominal side of scrotum or perineum[1-2]It is one of the common malformations of human genitourinary system. The early and middle stage of pregnancy is difficult to diagnose, and the physician who is in clinical consultation after the birth of the baby can determine whether the baby has hypospadias by observing whether the abnormal opening of the urethra exists or not with naked eyes[1]. Therefore, the clinical urgent need is to develop the early diagnosis research of hypospadias, and the current combined examination technology of maternal serum AFP and free beta-HCG is widely applied to the prenatal screening in the fields of screening 21 and 18 trisomy syndromes of fetus, open neural tube defect, abdominal fissure and umbilical bulge[18-20]However, the use of maternal AFP in combination with free β -HCG for the prediction of fetal hypospadias is rarely reported. Therefore, the present study was conducted mainly on the correlation and diagnostic value of AFP and free β -HCG levels with hypospadias fetuses.
The research result shows that the level of free beta-HCG MOM in the pregnant woman in the group of the hypospadias of the fetus is higher than that in the control group, and the difference has statistical significance (P is less than 0.05). The AUC of the free beta-HCG screened hypospadias is 0.659 (95% CI: 0.565-0.752, P ═ 0.002). When the optimal cutoff value of free beta-HCG for screening the hypospadias is 1.275MOM, the sensitivity, specificity and Jordan index are respectively as follows: 0.522, 0.790 and 0.312. And Schneuer et al[21]The research result shows that the serum free beta-HCG level of the pregnant woman in New Nanwelshizhou with hypospadias is 0.88 (0.66-1.40) MOM to 0.92 (0.65-1.38) MOM, P is 0.83; the Western Australia state pregnant women are 0.84 (0.63-1.28) MOM to 0.88 (0.59-1.30) MOM, P is 0.76, and the pregnant women in the 2 groups have no statistical significance in difference compared with the normal control group (P is more than 0.05). The research result indicates that the serum free beta-HCG level of the maternal urine at the early pregnancy period has no correlation with the hypospadia or the cryptorchidism of the fetus, which is opposite to the research result.
The AFPMOM level in the study case group was higher than that in the control group, and the difference was statistically significant (P)<0.05). AUC of AFP screening hypospadias is 0.644 (95% CI: 0.550-0.737, P ═ 0.005). When the optimal cutoff value of AFP singly screening the hypospadias is 0.945MOM, the sensitivity, specificity and Youden index at the time are respectively as follows: 0.739, 0.484, 0.223. Papp et al[22]Studies have shown that with AFP ≧ 2.5MOM as the cutoff value, only 12.8% (5/39) of the hypospadias were detected by maternal serum AFP MOM levels. The study was carried out with AFP ≧ 2.5MOM
Statistically, only 4 of 69 hypospadias were AFP ≧ 2.5MOM with sensitivity of 5.80% (4/69), which is lower than the results of the Papp study. Therefore, it is proposed to implement localized AFP MOM cut-off values to improve the detection sensitivity and specificity of AFP for predicting a hypospadias fetus.
The research result also shows that the AUC of the fetal hypospadias predicted by the combination of AFP + free beta-HCG is 0.700 at the highest, which is higher than 0.644 of AFP or 0.659 of free beta-HCG, and the sensitivity of the fetal hypospadias predicted by the combination of AFP + free beta-HCG is 0.551, lower than 0.739 of AFP and higher than 0.522 of free beta-HCG; however, the specificity was 0.855, which is higher than both 0.484 and 0.790. Therefore, the method has higher diagnostic value by predicting the fetal hypospadias according to the age of the pregnant women and the combination of AFP and free beta-HCG of serum.
Table 1 comparison of baseline data for each group
TABLE 2 comparison of Hyposparias with serum AFP, free beta-HCG levels during pregnancy in control groups
TABLE 3 AFP, free beta-HCG diagnosis Hypospias values alone and in combination
Reference to the literature
[1] Shao chuanmei, leaf hong bei, qiu xiaoshan, practical neonatology (4 editions) [ M ]. people's health publishing agency, beijing: in 2011: 663.
[2]Vand ZLFM,Van Rooij IALM,Feitz WFJ,et al.Aetiology of hypospadias:a systematic review of genes and environment[J].Human Reproduction Update,2012,18(3):260-283.
[3]Geller F,Feenstra B,Carstensen L,et al.Genome-wide association analyses identify variants in developmental genes associated with hypospadias[J].Nature Genetics,2014,46(9):957-963.
[4]Kurahashi N.Maternal genetic polymorphisms in CYP1A1,GSTM1 and GSTT1 and the risk of hypospadias[J].Molecular Human Reproduction,2005,11(2):93-98.
[5]Jen SA,Xian NQQ,Ang WW.Association of Prenatal Exposures of PolybrominatedDiphenyl Ethers With Hypospadias[J].JAMA Pediatr.2018,(19):
[6]GermaniM,PérezMDF,SoleraLS.Endocrine disruptors and hypospadias in Gran Canaria island(2012-2015)[J].Rev EspSalud Publica.2018Aug 29;92
[7]Elodie H,Pierre T,Christelle K,et al.Isolated hypospadias:The impact of prenatal exposure to pesticides,as determined by meconium analysis[J].Environment International,2018,119(1):20-25.
[8] mutation study of gene related to chentaiji-hypospadiae [ D ]. university of china cooperative medical science, 2006.
[9] The expression and significance of FGFR2 in foreskin tissue of hypospadias patients [ D ]. university of south and middle, 2007.
[10]Diamond DA,Chan I,Holland A,et al.Advances in paediatric urology[J].Lancet,2017,390(10099):1061-1071.
[11] Chenyougan root, MAMLD1, SF-1 gene and hypospadias relationship study [ D ]. Shandong university, 2010.
[12]Mohammadzadeh A,Farhat A,Esmaieli H,et al.Prevalence and risk factors of hypospadias in a private hospital in northeast iran[J].Iranian Journal of Pediatrics,2011,21(4):497-501.
[13] Chinese birth defect monitoring scheme chinese maternal-child hygiene monitoring workbook (2013 edition) [ R ]. national maternal-child hygiene monitoring and yearbook communication, beijing: department of health, women and children health and community health department, 2013, 50 (1): 220.
[14] sincerity, liu nationality, jinhuiming, etc. neonate's science [ M ]. people's health press, 2002: 636-637.
[15]Cuckle H S,Wald N J,Thompson S G.Estimating a woman's risk of having a pregnancy associated with Down's syndrome using her age and serum alpha-f etoprotein level[J].BJOG:An International Journal of Obstetrics&Gynaec ology,1987,94(5):387-402.
[16]Royston P,Thompson S G.Model-based screening by risk with application to down's syndrome[J].Statistics in medicine,1992,11(2):257-268.
[17] chen Kun, Chen Zhong. medical research methods [ M ]. Beijing: Science Press,2011:43-44.(in Chinese) Chenkun, Chen faithful. method of scientific research [ M ]. Beijing: scientific press,2011:43-44.
[18] Chen Yiming, Zhang Wen, Wang Hao, etc. study of maternal serum AFP and free beta-HCG subunit in the middle pregnancy to screen fetal chromosomal aneuploidy abnormality [ J ] Chinese journal of preventive medicine, 2018, 19(1):13-17.
[19] Screening results of maternal serum AFP and free beta-HCG in pregnancy in fetal neural tube malformation [ J ] Chinese public health, 2018, 34(6):854 and 857.
[20] Screening efficiency of fetal abdominal fissure and umbilical cord expansion by maternal serum alpha fetoprotein and free beta-hCG in middle and pregnancy (J) university of Zhejiang (medical edition), 2017,46(3) 268 and 273.
[21]Schneuer FJ,Bower C,Holland AJA,et al.Maternal first trimester serum levels of free-beta human chorionic gonadotrophin and male genital anomalies[J].Human Reproduction,2016,31(8):1895-1903.
[22]Papp Z,E.Tóh-Pál,Papp C,et al.Impact of prenatal mid-trimester screening on the prevalence of fetal structural anomalies:a prospective epidemiological study[J].Ultrasound in Obstetrics and Gynecology,1995,6(5):320-326.
It should be understood that equivalent substitutions and changes to the technical solution and the inventive concept of the present invention should be made by those skilled in the art to the protection scope of the appended claims.
Claims (1)
1. A method for establishing a prediction risk model of a maternal serum marker for fetal hypospadias is characterized by comprising the following steps: the method comprises the following steps:
(1) dividing the pregnant women into a case group and a control group according to the existence of the hypospadias of the fetus, wherein the case group is the pregnant women who are clinically confirmed to give birth to the hypospadias of the fetus, and the control group is the pregnant women who randomly draw the fetus in the same period and have normal development;
(2) detecting the AFP and free beta-HCG levels of the 2 groups of pregnant woman serum by adopting a time-resolved immunofluorescence method, constructing a risk calculation model by using the AFP and free beta-HCG, and determining an optimal cutoff value and an area AUC under a curve according to an ROC curve;
(3) during detection, detecting the AFP and free beta-HCG levels of the serum of the pregnant woman to be detected by adopting a time-resolved immunofluorescence method, screening by using a risk calculation model constructed by the AFP and free beta-HCG, and judging the fetus of the pregnant woman to be detected to be a hypospadias fetus when the AFP and free beta-HCG levels of the serum of the pregnant woman to be detected exceed a set threshold value;
the risk calculation model is: the Lifecycle-like risk value is calculated by multiplying the age risk value by the corresponding normal distribution likelihood ratio value,
the age equation: riskage=0.000627+exp-16.2395+0.286*(age-0.5)
Wherein riskageIs age risk value, age is pre-delivery age
two-dimensional normal distribution likelihood calculation formula:
wherein sigma is the standard deviation of the corresponding index, rho is the correlation coefficient of the two indexes, mu is the mean value of the sample, where x denotes the logarithm of AFP MoM value, y is the logarithm of free-beta HCGMoM value, and the final hypospadias risk value:
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