Background: Paired-like Homeobox 2B (PHOX2B) is considered the causative gene of Congenital Central Hypoventilation Syndrome (CCHS), a dominant genetic disorder characterized by impaired central respiratory control and subsequent hypoventilation during sleep. Methods: Herein, we present a family with recurrent severe CCHS. The potential causative genetic variant was confirmed through Whole-Exome Sequencing (WES), Sanger sequencing, and droplet digital PCR (ddPCR). Furthermore, prenatal diagnosis was performed on the proband's mother at 20 weeks of her fourth pregnancy upon request. Results: The proband and her brother were both carriers of the PHOX2B polyalanine expansion variant: c.744_758dupCGCGGCAGCGGCGGCGGCGGC. Sanger sequencing revealed that the proband's father had a small variant peak in the gene position, implying potential somatic mosaicism. In addition, ddPCR results showed that the proband's father had germline mosaicism, with a mosaicism proportion of 14.3%. Notably, the detect p. (Ala241[26]) variant was not detected in the fetus. Conclusions: These findings have important implications for improving genetic counseling of CCHS families as they suggest that even parents without CCHS symptoms may have somatic chimerism, necessitating careful genetic counseling and consideration of prenatal testing for subsequent pregnancies.

Background: Preeclampsia (PE) is an idiopathic disorder of pregnancy. The exact cause of PE remains unknown. Emerging evidence indicates that the cause of PE is linked to genetic factors. Therefore, the aim of this study was to identify the susceptibility genes for PE.Methods: Nine families with severe PE were recruited. The whole-exome sequencing (WES) was performed on each family, and Sanger sequencing was used to identify the potential pathogenic genetic variants.Results: After a rigorous bioinformatics analysis, compound heterozygous variants in the NPFFR2 gene, NM_004885.2: c.601A > G, p.Met201Val and c.995C > T, p.Ala332Val were found in the No.4 pedigree. Bio-informatics analysis showed that these sites were highly conserved among several species and were predicted to be pathogenic variants according to multiple online mutational function prediction software packages. Due to the compound heterozygous variants of NPFFR2, more bonds are generated between mutant amino acids and spatial adjacent amino acids, which may lead to more stable active conformation of protein and not easy to be degraded.Conclusions: We demonstrated for the first time that compound heterozygous variants of the NPFFR2 gene might be potentially associated with severe PE, the results of this study provide clinicians and researchers with a better understanding of the molecular mechanisms underlying severe PE in pregnant women.

OBJECTIVE:To study the molecular epidemiology of thalassemia in Jiaxing area of Zhejiang province and provide a basis for prenatal diagnosis, genetic counseling and prevention and control of birth defects.METHODS:A total of 24 003 pregnant women who presented at the Jiaxing Maternal and Child Health Care Hospital from April 2017 to September 2021 were enrolled. Capillary hemoglobin electrophoresis in combination with routine blood test were used for primary screening for carriers of thalassemia-associated mutations, and those with positive results were subjected to fluorescence quantitative PCR assay. Prenatal diagnosis was provided for couples with a risk of giving birth to children with intermediate or severe thalassemia.RESULTS:Among the 24 003 pregnant women, 1 211 cases were suspected as carriers of thalassemia-associated mutations, among whom 443 (36.58%) were confirmed by genetic testing. Among these, carriers of α-, β- and α-complex β-globin gene mutations have accounted for 27.31% (121/443), 70.65% (313/443) and 2.04% (9/443), respectively. The result of prenatal diagnosis for an at-risk couple was --SEA/αCSα, and the fetus was predicted to have intermediate or severe thalassemia. Termination of the pregnancy was recommended.CONCLUSION:Hemoglobin electrophoresis combined with routine blood test during pregnancy may be used as a preliminary screening measure for carriers of thalassemia-associated variants. Combined with genetic testing, this will be of great significance for the control of thalassemia in this region.

BACKGROUND:Preeclampsia (PE), a pregnancy specific syndrome, is one kind of common gestational hypertension disease, which can cause maternal and perinatal mortality and morbidity. This study was conducted to identify key microRNAs (miRNAs), mRNAs and related signaling pathways in the pathogenesis of PE.METHODS:Whole transcriptome sequencing and small RNA sequencing of the peripheral blood from 3 PE patients and 3 normal pregnant women were performed. Differential expressed (DE) miRNAs were identified using the DEseq2 package. Target genes of the selected upregulated and downregulated DE miRNAs were predicted. Based on the hypergeometric distribution of DE miRNA target genes, we analyzed GO enrichment and KEGG pathway enrichment using R.RESULTS:Total 1291 and 1281 novel RNAs were obtained from the preeclampsia patients and healthy individuals. 70 miRNAs were screened out with significant levels with 51 significantly upregulated and 19 significantly downregulated. 44,306 genes were predicted as the targets of these miRNAs. Besides, KEGG pathway analysis revealed that the upregulated miRNAs were enriched in Glycosaminoglycan biosynthesis-chondroitin sulfate / dermatan sulfate, Base excision repair and the downregulated miRNAs were enriched in Tuberculosis, Phagosome.CONCLUSION:We constructed regulatory networks of miRNAs and target genes, there were 2208 negative miRNA-mRNA interactions in total. The network and pathway information illustrate the potential functions of mRNAs and miRNAs in PE pathogenesis.

目的 对一个生育过脊髓性肌萎缩症患儿的家系进行产前诊断,同时对三种实验室检测技术进行方法学的比较.方法 应用荧光定量PCR、荧光PCR-毛细管电泳法及MLPA对家系外周血样本及绒毛组织、羊水组织进行了检测.同时进行了亲缘鉴定以排除母源污染.结果 第一胎患儿为SMN1基因7、8号外显子纯合缺失合并SMN2基因的3拷贝重复,患儿父亲为SMN1基因第7、8号外显子的杂合缺失携带者合并SMN2基因的3拷贝重复,患儿母亲为SMN1基因第7、8号外显子的杂合缺失携带者,SMN2基因2拷贝.本次妊娠胎儿为SMN1基因第7、8号外显子杂合缺失及SMN2基因3拷贝重复.结论 经检测胎儿仅为SMN1基因杂合缺失的携带者,可继续正常妊娠.同时,荧光PCR-毛细管电泳法和MLPA可同时对SMN1和SMN2基因拷贝数进行检测,检测较为全面,但荧光定量PCR检测操作方便,检测周期短,适合大样本人群的普筛.

Background Molecular genetic testing for the 11p15-associated imprinting disorder Beckwith-Wiedemann syndrome (BWS) is challenging because of the molecular heterogeneity and complexity of the affected imprinted regions. An integrated molecular approach to analyze the epigenetic-genetic alterations is required for accurate diagnosis of BWS. Case presentation : We reported a Chinese case with BWS detected by SNP array analysis and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA). The genetic analysis showed a de novo duplication of 24 Mb at 11p15.5p14.3 is much longer than ever reported. MS-MLPA showed copy number changes with a peak height ratio value of 1.5 (three copies) at 11p15. The duplication of paternal origin with increase of methylation index of 0.68 at H19 and decreased methylation index of 0.37 at KCNQ1OT1 . Conclusion Combined chromosome microarray analysis and methylation profiling provided reliable diagnosis for this paternally derived duplication of BWS. The phenotype associated with 11p15 duplications depends on the size, genetic content, parental inheritance and imprinting status. Identification of these rare duplications is crucial for genetic counselling.

目的:分析本院新生儿耳聋基因筛查的结果,明确嘉兴地区新生儿耳聋基因的突变情况.方法:选取2018年3月-2019年10月在本院出生的新生儿,根据知情自愿原则采集8111例新生儿的脐带血或足跟血(1～2 ml),应用基因芯片技术对中国人常见4个耳聋基因共9个突变位点[GJB2(235delC、35delG、299delAT、176del16),GJB3(538C>T),SLC26A4(IVS7-2A>G、2168A>G),线粒体12SrRNA(1555A>G、1494C>T)]进行筛查.结果:8111例本院出生的新生儿中共筛查出397例至少携带1个耳聋基因突变,突变率4.895％,其中GJB2基因突变185例,突变率为2.281％;GJB3基因突变22例,突变率0.271％;SLC26A4基因突变146例,突变率1.800％,线粒体12SrRNA基因突变37例,突变率0.456％,双杂合突变7例.结论:常见耳聋基因在嘉兴地区新生儿中有较高的阳性率,主要以GJB2基因的235delC、SLC26A4基因的IVS7-2A>G为主.建议有条件的地区全面开展新生儿耳聋基因的筛查,有助于遗传性耳聋、药物性耳聋和迟发性耳聋的早发现、早诊断和早治疗.

Abstract Background: Congenital heart disease (CHD) is an important birth defect, but its mechanism is still unclear. In recent years, genetic causes including chromosomal abnormalities are associated with the occurrence of congenital heart disease. In this study, CMA technology is applied to explore the genetic causes of congenital heart disease, so as to further clarify the correlation between genotype and phenotype and prepare for late pregnancy intervention and postnatal diagnosis and treatment.Objective: To explore the chromosomal abnormalities and copy number variation (CNVs) of fetuses with CHD by CMA technology, and to clarify the clinical application value of CMA technology as a detection method of first-tier antenatal CHD.Methods: Amniotic fluid sample from 155 pregnant women diagnosed with fetus CHD by prenatal ultrasound from 2018 to 2021 are collected for SNP-array detection and karyotype analysis. According to the detected CNVs results, FISH, CMA or karyotype analysis are further selected for parental verification.Results: Among the 155 fetuses with CHD, a total of 32 (20.6%) cases of chromosomal abnormalities are detected, of which 31.3% are chromosome number abnormalities. CNVs of likely pathogenicity and unknown significance are 2.5% and 5.2% respectively. The detection rate of chromosomal abnormalities in CHD of different subtypes is different, among which the high detection rate is complex CHD (31.2%), right ventricular outflow tract obstruction (30.7%) and conotruncal defects (25%). The detection rate of chromosomal abnormalities in CHD with extracardiac structural abnormalities is significantly higher than that in isolated CHD (52.4% vs 11.3%, p<0.05). In addition, the detection rate of CHD with abnormal extracardiac structure is significantly higher than that of CHD with soft markers (52.4% vs 17.8%, p<0.05), which is statistically significant. There is no significant difference in detection rate between CHD with soft markers and isolated CHD (17.8% vs 11.3%). Of the 155 pregnant women with fetus CHD, 59 chose to terminate their pregnancies, some of which were terminated according to the results of SNP-array, and some of which were terminated according to the severity of CHD.Conclusion: SNP-array technology can be used to detect chromosomal abnormalities of first-tier antenatal CHD fetuses, with high resolution, short reporting period and high efficiency. Meanwhile, pregnancy intervention can be taken according to the results.

Background Preeclampsia (PE) is a pregnancy-specific syndrome, belongs to the gestational hypertension diseases category and is considered among the causes of maternal and perinatal mortality and morbidity. However, the pathogenesis of PE is still vague. Methods In the present study, the circular RNA (circRNA) expression patterns of normal pregnant women and PE patients were investigated using whole RNA sequencing. Results A total of 151 differential expressed circRNAs were identified including 121 upregulated and 30 downregulated ones. Functional and pathway enrichment analysis was conducted on the differentially expressed circRNAs using Gene Ontology and KEGG databases. The results of this analysis indicated that several crucial biological processes and pathways were enriched in PE patients. circRNA–microRNA (miRNA) interaction analysis indicated that the reported differentially expresse circRNAs may be associated with some regulatory functions through miRNAs in PE patients. Two ceRNAs networks were constructed according to the targeting relationship between circRNAs/miRNAs and miRNAs/mRNAs. One sub-network contained one upregulated circRNA, four downregulated miRNAs and five upregulated mRNAs, and another sub-network contained 10 downregulated circRNAs, 21 upregulated miRNAs and 15 downregulated mRNAs. Conclusion CircRNA expression patterns have been investigated and this analysis revealed their potential regulatory mechanisms in PE patients. We constructed the ceRNAs (competing endogenous RNA) to reveal the potential molecular roles of dysregulated circRNAs in the PE patients using RNA sequencing data. circRNA_13301 was the only one upregulated circRNA in ceRNA being targeted by four miRNAs.

Background .Preeclampsia (PE) is a pregnancy-specific syndrome, belongs to the gestational hypertension diseases category and is considered among the causes of maternal and perinatal mortality and morbidity.However, the pathogenesis of PE is still vague. Methods.In the present study, the circular RNA (circRNA) expression patterns of normal pregnant women and PE patients were investigated using whole RNA sequencing.Results.151 differential expressed circRNAs were identified including 121 upregulated and 30 downregulated ones.Functional and pathway enrichment analysis was conducted on the differentially expressed circRNAs using Gene Ontology and KEGG databases.The results of this analysis indicated that several crucial biological processes and pathways were enriched in PE patients.circRNA-microRNA (miRNA) interaction analysis indicated that the reported differentially expresse circRNAs may be associated with some regulatory functions through miRNAs in PE patients.Two ceRNAs networks were constructed according to the targeting relationship between circRNAs/miRNAs and miRNAs/mRNAs.One sub-network contained one upregulated circRNA, four downregulated miRNAs and five upregulated mRNAs, and another sub-network contained 10 downregulated circRNAs, 21 upregulated miRNAs and 15 downregulated mRNAs. Conclusion .CircRNA expression patterns have been investigated and this analysis revealed their potential regulatory mechanisms in PE patients.We constructed the ceRNAs (competing endogenous RNA) to reveal the potential molecular roles of dysregulated circRNAs in the PE patients using RNA sequencing data.circRNA_13301 was the only one upregulated circRNA in ceRNA being targeted by four miRNAs.

Background .Preeclampsia (PE) is a pregnancy-specific syndrome, belongs to the gestational hypertension diseases category and is considered among the causes of maternal and perinatal mortality and morbidity.However, the pathogenesis of PE is still vague. Methods.In the present study, the circular RNA (circRNA) expression patterns of normal pregnant women and PE patients were investigated using whole RNA sequencing.Results.151 differential expressed circRNAs were identified including 121 upregulated and 30 downregulated ones.Functional and pathway enrichment analysis was conducted on the differentially expressed circRNAs using Gene Ontology and KEGG databases.The results of this analysis indicated that several crucial biological processes and pathways were enriched in PE patients.circRNA-microRNA (miRNA) interaction analysis indicated that the reported differentially expresse circRNAs may be associated with some regulatory functions through miRNAs in PE patients.Two ceRNAs networks were constructed according to the targeting relationship between circRNAs/miRNAs and miRNAs/mRNAs.One sub-network contained one upregulated circRNA, four downregulated miRNAs and five upregulated mRNAs, and another sub-network contained 10 downregulated circRNAs, 21 upregulated miRNAs and 15 downregulated mRNAs. Conclusion .CircRNA expression patterns have been investigated and this analysis revealed their potential regulatory mechanisms in PE patients.We constructed the ceRNAs (competing endogenous RNA) to reveal the potential molecular roles of dysregulated circRNAs in the PE patients using RNA sequencing data.circRNA_13301 was the only one upregulated circRNA in ceRNA being targeted by four miRNAs.

目的 针对嘉兴地区15 574名新生儿群体进行中国人群常见耳聋基因突变的分子筛查,分析嘉兴地区人群耳聋基因突变携带情况.方法 新生儿分娩时,采集新生儿脐带血2-3ml,采用PCR与导流杂交法相结合技术,检测中国人群中最常见的4个耳聋致病基因上的9个突变位点.并对复合杂合突变及双杂合突变和纯合突变的样本进行了随访.结果 15 574例受检新生儿中共有696例(4.47％)携带至少1个耳聋基因突变.其中GJB2基因杂合突变363例,占2.33％;GJB3基因杂合突变的36例,占0.23％;SLC26A4基因杂合突变233例,占1.50％;线粒体12SrRNA基因47例,占0.30％;携带有两个耳聋基因位点突变的有13例.目前听力语言发育正常.另外,存在GJB2基因c.235delC、c.176-191de116复合杂合突变1例(0.64‰),GJB2基因c.235delC纯合突变2例(1.28‰).SLC26A4基因c.919-2A＞G纯合突变1例(0.64‰).出生时、三个月时的听力筛查双耳均未通过,目前仍在继续随访.结论 本研究对嘉兴地区人群遗传性耳聋热点突变筛查进行大样本的探索,丰富了非综合征耳聋基因突变谱,为耳聋基因突变筛查、耳聋基因诊断及遗传咨询提供了重要数据资料.从而进行早发现、早干预、早治疗,提高聋儿的生存质量,并对耳聋基因突变携带者进行婚育指导,降低本地区出生缺陷.

目的:探讨将细菌人工染色体微球(Bobs)检测技术应用于胎儿染色体异常产前诊断的可行性.方法:选取7864例单胎中期妊娠孕妇(指征包括高龄、唐氏血清学筛查高风险、胎儿无创DNA检测高风险、不良生育史、胎儿超声结构或软指标异常、染色体病家族史等),用Bobs技术快速检测13、18、21、X、Y染色体的数目异常及9种微缺失综合征,并与常规羊水细胞染色体的G显带分析结果及基因芯片检测结果进行比较.结果:共检出302例各类异常,包括256例染色体非整倍体数目异常(含3例嵌合体及2例47,XXY合并Yq11.223、Yq11.23微重复),46例微缺失及微重复异常(含1例等臂X染色体),染色体非整倍体数目异常检测结果同G显带核型分析结果一致,有39例微缺失、微重复进行基因芯片检测,结果与Bobs一致.Bobs在检测数目异常十嵌合体及结构异常的灵敏度、特异性和准确性均为100.0％.结论:Bobs技术检测胎儿染色体非整倍体异常和9种微缺失综合征,是具有快速、可靠等特点的产前诊断方法.

目的 研究产钳助产术对产后出血量的影响及其原因分析.方法 回顾性分析我院2016年167例行产钳助产术产妇的产后出血量,分别与175例自然分娩产妇和51例经阴道分娩过程中行会阴侧切术产妇的产后出血量进行比较.同时针对行产钳助产术的三病因(6例胎位不正,118例胎儿宫内窘迫以及43例第二产程延长)分别与自然分娩产妇的出血量相比较.结果 产钳助产组的平均产后出血量为(294.97±84.28)ml,较自然分娩组的平均产后出血量(242.17±56.96)ml多,两组之间具有显著性差异(P＜0.05).同时,产钳助产组出血量较会阴侧切组(245±46.41mL)多,具有统计学差异(P＜0.05).另外,产钳助产组三病因的平均出血量(胎位不正组(316.67±68.31)mL,胎儿宫内窘迫组(285.59±60.63) ml,第二产程延长组(317.67±128.21) mL均较自然分娩组多,存在统计学差异.结论 产钳助产对产后出血量的增加提示产钳助产本身或产钳运用的病因可能会增加产后出血,但产钳助产对困难分娩依然有着其独特的优势.

OBJECTIVE:To determine the origin of a supernumerary small marker chromosome found in a fetus using prenatal BACs-on-Beads (BoBs) and single nucleotide polymorphism array (SNP-array) assays.METHODS:The fetal sample was subjected to chromosomal karyotyping and BoBs analysis, and the results were validated with genome-wide scanning using a SNP microarray.RESULTS:The fetus was found to have a 47,XX,+mar karyotype. BoBs analysis indicated that there was an amplification between 18p11.32 and 18p11.21, which was verified by the SNP-array assay as a 18.3 Mb duplication occurring at 18p11.32q11.1.CONCLUSION:The karyotype of the fetus was determined as 47,XX,+der18(18p11.32?18q11.1::18q11.1?18p11.32). The duplication has involved important genes including SMCHD1, LPIN2 and TGIF1, which may result in severe malformations in the fetus.

随着2型多发性内分泌腺瘤(multiple endocrine neoplasia type 2,MEN 2)不同临床症状与RET原癌基因不同突变型相关研究的发现,以及更多的肿瘤靶向药物运用于临床试验,使得MEN 2的早期确诊和药物治疗成为可能.对于无法实施手术的重症MEN 2患者,Vandetanib和Carbozantinib等分子靶向药物可以起到一定的疗效.本文介绍了MEN 2的常规检测与遗传学诊断,未来MEN 2的治疗研究方向,将PI3K/Akt/mTOR信号通路和CRISPR/TALEN-Cas9等知识运用到MEN 2的非手术治疗中的可能性,以及通过产前诊断和植入前遗传学诊断有效避免患病胎儿的出生.

Epidermolytic palmoplantar keratoderma (EPPK) is the most frequent form of such keratodermas. It is inherited in an autosomal dominant pattern and is clinically characterized by diffuse yellowish thickening of the skin on the palms and soles with erythematous borders during the first weeks or months after birth. EPPK is generally caused by mutations of the KRT9 gene. More than 26 KRT9 gene mutations responsible for EPPK have been described (Human Intermediate Filament Database, www.interfil.org), and many of these variants are located within the highly-conserved coil 1A region of the α-helical rod domain of keratin 9. Unfortunately, there is no satisfactory treatment for EPPK. Thus, prenatal molecular diagnosis or pre-pregnancy diagnosis is crucial and benefits those affected who seek healthy descendants. In the present study, we performed amniotic fluid-DNA-based prenatal testing for three at-risk pregnant EPPK women from three unrelated southern Chinese families who carried the KRT9 missense mutations p.Arg163Trp and p.Arg163Gln, and successfully helped two families to bear normal daughters. We suggest that before the successful application of preimplantation genetic diagnosis (PGD), and noninvasive prenatal diagnosis of EPPK that analyzes fetal cells or cell-free DNA in maternal blood, prenatal genetic diagnosis by amniocentesis or chorionic villus sampling (CVS) offers a quite acceptable option for EPPK couples-at-risk to avoid the birth of affected offspring, especially in low- and middle-income countries.

BACKGROUND:Genetic screening for germline mutations in the RET proto-oncogene has been extensively exploited worldwide to optimize the diagnostic and clinical management of multiple endocrine neoplasia type 2 (MEN2) patients and their relatives. However, a distinct lag period exists not only in the recognition but also in the medical treatment of patients with MEN2. Here we present a comprehensive genetic and clinical analysis of MEN2 among Chinese families followed from 1975 to 2011. Our series comprises 36 index cases and 134 relatives from 11 independent families.METHODS:Genetic diagnosis was performed in all participants by direct sequencing all relevant RET exons. Thyroidectomy was performed in 50 patients with varying cervical neck dissection procedures. Patients with pheochromocytoma (PHEO) underwent specific surgery. Demographic, clinical profiles, mutation types, tumor histopathologic features, and follow-up records were systematically analyzed.RESULTS:The RET mutations p.C634Y (n=34), p.C634R (n=6), p.C618S (n=13), p.V292M/R67H/R982C (n=7), p.L790F (n=2), and p.C634Y/V292M/R67H/R982C (n=1) were confirmed in 31 index cases and then identified in 32 at-risk relatives (mutation carriers), with MEN2A as the most common clinical subtype. The overall penetrance of PHEO in patients with MEN2A was 46.7%. A total of 50 patients underwent thyroidectomy, and there was a significant lowering of their mean age at thyroidectomy and the tumor diameter of the mutation carriers that were detected and operated on compared with the index cases (age at first surgery: 29.3 vs. 39.3 years, p<0.05; maximum size: 1.1 vs. 3.3 cm, p<0.001). There was also a decrease in the TNM staging and the proportion of patients who underwent inappropriate initial thyroid surgery (pN1: 31.6% vs. 100%, p<0.001; inappropriate surgery: 0% vs. 29%). Meanwhile, disease-free survival (DFS) increased (DFS: 100% vs. 58.1%, p<0.05). Both medullary thyroid carcinoma-specific (n=1) and PHEO-specific (n=5) deaths were reported during the study period.CONCLUSIONS:Our results further substantiate that gene scanning of all relevant RET exons is a powerful tool in the management of MEN2 patients, especially in asymptomatic carriers, and has led to earlier diagnosis and more complete initial treatment of patients with MEN2 in China.

Mutation-based molecular diagnostics of autosomal dominant polycystic kidney disease (ADPKD) is complicated by genetic and allelic heterogeneity, large multi-exon genes, and duplication sequences of PKD1. Recently, targeted resequencing by pooling long-range polymerase chain reaction (LR-PCR) amplicons has been used in the identification of mutations in ADPKD. Despite its high sensitivity, specificity and accuracy, LR-PCR is still complicated. We performed whole-exome sequencing on two unrelated typical Chinese ADPKD probands and evaluated the effectiveness of this approach compared with Sanger sequencing. Meanwhile, we performed targeted gene and next-generation sequencing (targeted DNA-HiSeq) on 8 individuals (1 patient from one family, 5 patients and 2 normal individuals from another family). Both whole-exome sequencing and targeted DNA-HiSeq confirmed c.11364delC (p.H3788QfsX37) within the unduplicated region of PKD1 in one proband; in the other family, targeted DNA-HiSeq identified a small insertion, c.401_402insG (p.V134VfsX79), in PKD2. These methods do not overcome the screening complexity of homology. However, the true positives of variants confirmed by targeted gene and next-generation sequencing were 69.4%, 50% and 100% without a false positive in the whole coding region and the duplicated and unduplicated regions, which indicated that the screening accuracy of PKD1 and PKD2 can be largely improved by using a greater sequencing depth and elaborate design of the capture probe.