OBJECTIVE:To characterize prenatal phenotypes and genetic basis of ciliopathies identified by prenatal exome sequencing (pES) in fetuses with ultrasound abnormalities. METHOD:Singleton pregnancies who underwent pES due to fetal ultrasound abnormalities were retrospectively reviewed between 2020 and 2023. Cases affected by ciliopathies were included if diagnostic variants were identified in 959 ciliopathy-associated gene entries in the manually curated SCGSv2 and integrated database CilioGenics (top 500). Phenotype-genotype correlations were analyzed. RESULTS:A total of 41 cases were diagnosed as ciliopathies by pES with a detection rate of 7.4% (41/555). DYNC2H1 and FGFR3 were the most common genes associated with first-order and second-order skeletal ciliopathies, while PKD1 was the major causative gene of renal ciliopathies. Consistent with genetic diagnoses, skeletal (53.7%, 22/41) and kidney (34.1%, 14/41) abnormalities were frequent features by ultrasound. Kidney abnormalities (59.1%, 13/22) were more prominent in first-order ciliopathies, especially hyperechogenic kidneys. By contrast, skeletal findings (78.9%, 15/19) were the leading ultrasound sign in second-order ciliopathies, usually manifesting as short limbs. CONCLUSIONS:Ciliopathies prenatally present with variable ultrasound features, with kidney and skeletal systems frequently affected in first-order and second-order ciliopathies. Characterization of the genetic etiology and phenotypes of prenatal ciliopathies will increase our understanding of fetal genetic disorders.

OBJECTIVE:To explore the genetic etiology of a Chinese pedigree affected with Branchio-oculo-facial syndrome (BOFS) and summarize the prenatal phenotype of BOFS patients. METHODS:A pedigree with BOFS which had presented at the Genetics and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University in December 2021 was selected as the study subject. Clinical data of the pedigree was collected. The fetus was subjected to routine prenatal ultrasound scan. Trio-whole exome sequencing (trio-WES) was carried out for the fetus and its parents, and candidate variant was verified by Sanger sequencing. Relevant literature was searched from the database to summarize the prenatal phenotype of BOFS patients. RESULTS:Ultrasound exam suggested the fetus had cleft lip and palate. Its father had presented with high palatal arch, prematurely grayed hair, occult cleft lip, congenital preauricular fistula, red-green color blindness and unilateral renal agenesis. Its grandfather also had high palatal arch, prematurely gray hair, protruding ears, congenital preauricular fistula and hearing disorders. Trio-WES revealed that the fetus and its father had both harbored a heterozygous c.890-1G>A variant of the TFAP2A gene. The same variant was not found in its mother. Sanger sequencing confirmed that its grandfather had also harbored the same variant. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was rated as likely pathogenic (PVS1+PM2_Supporting). Combined with 36 similar cases retrieved from the literature, the prenatal phenotypes of BOFS patients had included growth restriction (25/37), renal abnormalities (10/37), cleft lip and palate (5/37) and oligohydramnios (5/37). CONCLUSION:The c.890-1G>A variant of the TFAP2A gene probably underlay the pathogenesis of BOFS in this pedigree. Discovery of the novel variant has enriched the mutational spectrum of the TFAP2A gene. The common prenatal phenotypes of BOFS have included growth restriction, renal abnormalities, cleft lip and palate and oligohydramnios. Delineation of the intrauterine phenotype of BOFS may facilitate its prenatal diagnosis, clinical diagnosis, treatment and genetic counseling.

ObjectiveWhole exome sequencing (WES) is becoming more widely used as a diagnostic tool in the field of medicine. In this article, we reported the diagnostic yield of WES and mitochondrial genome assessment in 2226 consecutive cases in a single clinical laboratory.Materials and methodsWe retrospectively analyzed consecutive WES reports from 2226 patients with various genetic disorders. WES- process was focused exclusively on the probands and aimed at a higher diagnostic capacity. We determined the diagnostic rate of WES overall and by phenotypic category, mode of inheritance, mitochondrial genome variant, and copy number variants (CNVs).ResultsAmong the 2226 patients who had diagnostic WES proband-only, the overall diagnostic yield of WES was 34.59% (770/2226). The highest diagnostic yield was observed in autosomal dominant disorders, at 45.58% (351/770), followed by autosomal recessive at 31.95%(246/770), X-linked disorder at 9.61%(74/770), and mitochondrial diseases at a notably lower 0.65%(5/770). The 12.21% (94/770) diagnoses were based on a total of 94 copy number variants reported from WES data. CNVs in children accounted for 67.02% of the total CNVs. While majority of the molecular diagnoses were related to nuclear genes, the inclusion of mitochondrial genome sequencing in the WES test contributed to five diagnoses. all mitochondrial diseases were identified in adults.ConclusionsThe proband-only WES provided a definitive molecular diagnosis for 34.59% of a large cohort of patients while analysis of WES simultaneously analyzed the SNVs, exons, mitochondrial genome, and CNVs, thereby improving the diagnostic yield significantly compared to the single-detection WES method; and facilitating the identification of novel candidate genes.

Background: The prenatal phenotype of neurological and neuromuscular monogenic diseases remains unclear. This study aimed to identify and explore prenatal ultrasonic presentation in fetuses with monogenic neurologic and neuromuscular diseases and to increase our knowledge on causal variants and novel candidate genes associated with fetal phenotypes. Methods: This retrospective study included 30 fetuses genetically diagnosed with or suspected of having neurologic and neuromuscular monogenic diseases who underwent prenatal trio-exome sequencing between January 2018 and May 2023. We systematically reviewed data on prenatal ultrasonic presentation, genetic characteristics, and genotype-phenotype relationships in a cohort of 30 fetuses. Some cases in this cohort study have been previously published as separate studies. Results: The disease spectrum of the 30 families included 27 diseases, including central nervous system diseases, neuromuscular diseases, congenital myopathy, mitochondrial diseases, and neurological genetic syndromes. The ultrasound abnormalities of the fetuses included abnormal fetal gestures and movements(18/30), polyhydramnios (15/30), increased nuchal translucency or nuchal fold (11/30), dysmorphic facial features (8/30), and brain malformations (6/30). Of the 40 variants detected, 35 were classified as pathogenic or likely pathogenic and 5 were of undetermined significance according to the American College of Medical Genetics and Genomics guidelines. The inheritance patterns were autosomal dominant (17/30; 57%), autosomal recessive (10/30; 33%), and X-linked recessive (3/30; 10%). Thirteen (13/30; 43%) fetuses achieved a definite genetic diagnosis, including known or novel variants of the recognized disease genes. Sixteen (16/30, 54%) fetuses were highly or probably suspected to be associated with genetic variants, and one fetus was found to have gene variants that did not explain the phenotype. Conclusions: The main clinical manifestations of prenatal ultrasonic presentation of neurological and neuromuscular monogenic diseases are abnormal fetal gestures, movements, and amniotic fluid volume, which might provide important clues for prenatal diagnosis. Trio-exome sequencing comprehensively detected and delineated various genomic variants that are causative. Our study expands our understanding of the prenatal phenotypic spectrum of monogenic neurological and neuromuscular diseases.

OBJECTIVE:To explore the genetic etiology of a child with delayed growth and development and carry out a literature review.METHODS:A child suspected for Al Kaissi syndrome at the First Affiliated Hospital of Zhengzhou University on March 6, 2021 was selected as the study subject. Following extraction of genomic DNA, the child was subjected to copy number variation sequencing (CNV-seq) and whole exome sequencing (WES), and candidate variants were verified by PCR-agarose gel electrophoresis and quantitative real-time PCR (qPCR). Prenatal diagnosis was conducted on chorionic villi sample upon subsequent pregnancy.RESULTS:The child, a 6-year-and-4-month-old boy, has dysmorphic features including low-set protruding ears and triangular face, delayed language and intellectual development, and ventricular septal defect. CNV-seq result has found no obvious abnormality, whilst WES revealed homozygous deletion of exons 1 and 2 of the CDK10 gene, which was confirmed by PCR-agarose gel electrophoresis and qPCR. Both of his parents were heterozygous carriers. Prenatal diagnosis using chorionic villi samples suggested that the fetus also carried the heterozygous deletion.CONCLUSION:The clinical features of Al Kaissi syndrome in this child can probably be attributed to the homozygous deletion of exons 1 and 2 of the CDK10 gene.

Objective To investigate the influence and reason analysis of the vanishing twin on noninvasive prenatal testing. Methods Clinical data of the pregnant women with vanishing twin syndrome (VTS) who came to our department for NIPT from January 2019 to December 2023 were collected, and their general conditions, NIPT results and prenatal diagnosis results as well as the causes of test failure were analyzed. The effects of gestational week of VTS and re-sampling time slot on the positive rate of NIPT were also discussed. Results Of the 102 cases, 5 cases were NIPT positive (4.9%), among them 4 cases were sex chromosome abnormality (SCA), and 1 case was ChrX-(Z=-7.38), that is, X chromosome reduction. The karyotype and CNV-seq results were all normal after amniocentesis. Namely, all the 5 cases were false positive, and the rate was 100% (5/5). The positive rate of VT < 10 weeks (3/73) was lower than that of VT ≥10 weeks (2/29). There was no significant difference in positive rate between the two groups. (P > 0.05) Conclusion NIPT could be applied on VT pregnancy, prolong the time interval of fetal demise and NIPT sampling. Once established, unnecessary invasive procedures could be avoided, mitigating negative emotional impact on future mothers.

OBJECTIVE:To assess the value of copy number variation sequencing (CNV-seq) for revealing the genetic etiology of fetuses with isolated ventricular septal defect (VSD).METHODS:From December 2017 to December 2020, 69 fetuses with isolated VSD were identified at the First Affiliated Hospital of Zhengzhou University. Meanwhile, 839 similar prenatal cases were selected from public databases including Wanfang data, Wanfang Medicine, and China National Knowledge Infrastructure (CNKI) by using keywords such as "Ventricular septal defect", "Copy number variation", and "Prenatal". A total of 908 fetuses with isolated VSD were analyzed. CNV-seq was carried out for 69 fetuses.RESULTS:Among the 908 fetuses, 33 (3.63%) were found to harbor pathogenic CNVs, which included 11 chromosomal aneuploidies (1.21%) and 22 pathogenic CNVs (2.42%). The pathogenic CNVs have involved 12 genetic syndromes, with those known to involve the heart development including 5 cases of 22q11.21 deletion syndrome, 2 cases of 4q terminal deletion syndrome, and 1 case of 9q subtelomere deletion syndrome. The outcome of pregnancies for 15 fetuses with pathogenic CNVs was known, of which 12 were terminated, and 3 had spontaneous closure of the ventricular septum after birth, but 1 of them had other abnormalities.CONCLUSION:Fetuses with isolated VSD have a relatively high risk for chromosomal abnormalities, for which CNV-seq should be recommended.

孕妇，女，25岁，G1P0，夫妻双方非近亲结婚，否认家族史、不良孕产史，平素身体健康，无不良嗜好，不吸烟酗酒，无不良接触史。于2020年11月(第1胎)停经16 + w进行孕中期常规产前超声检测，结果显示：双顶径2.8 cm，头围10.6 cm，腹围8.0 cm，股骨长0.6 cm，肱骨长0.6 cm。超声孕龄：14 +6 w。头部颅骨强回声环、大脑半球、脑中线、侧脑室、颅后窝池可见；双侧侧脑室脉络丛回声不均。脊柱朝后，显示不清。胸部心脏结构异常，腹部部分脏器、脐带腹壁入口可见；双上肢肱骨短小，双侧尺桡骨、指骨未见异常。超声诊断，四肢骨发育异常，考虑成骨发育不全的可能，心脏结构异常，左侧单脐动脉，双侧侧脑室脉络丛回声不均，考虑胎儿为多发畸形。建议引产并建议夫妻双方及胎儿做相关染色体及基因检测。征得孕妇知情同意及郑州大学第一附属医院伦理委员会批准(KS-2018-KY36)，引产后留取胎儿皮肤组织及父母外周血进行家系全外显子组检测。

Objective:To detect and analyze the gene variation types of 64 unrelated pedigrees affected with autosomal dominant polycystic kidney disease (ADPKD), and explore the detection efficiency of multiple gene analysis techniques and variation characteristics.Methods:It was a cross-sectional study. The clinical data of 64 pedigrees with ADPKD from Nephrology Department or Genetic and Prenatal Diagnosis Center of the First Affiliated Hospital of Zhengzhou University from December 2017 to August 2020 were retrospectively analyzed. The blood samples of probands and other family members were collected. Genetic analysis was carried out by next generation sequencing, and suspected mutations were verified by multiplex ligation-dependent probe amplification, or long-range PCR combined with Sanger sequencing. Prenatal diagnosis for high-risk fetuses was performed by fetal villi or amniotic fluid cells after genotyping without maternal genomic DNA contamination.Results:Among detected 64 pedigrees, 57 pedigrees (89.06%) had genetic variants in PKD1/PKD2. A total of 49 pathogenic/likely pathogenic variants in PKD1/PKD2 were identified in 51 pedigrees (79.69%), including 14 nonsense variants (28.57%), 14 frameshift variants (28.57%), 11 missense variants (22.45%), 5 splicing variants (10.20%) and 5 deletion variants (10.20%). Of these variants, 87.76% (43/49) were in PKD1 and 12.24% (6/49) were in PKD2. Totally, 14 novel variants in PKD1/ PKD2 were identified, including 7 frameshift variants, 3 splicing variants, 2 nonsense variants, 1 deletion variant and 1 missense variant, of which 11 variants were in PKD1 and 3 variants were in PKD2. Twenty high-risk fetuses from 17 pedigrees received prenatal diagnosis, in whom 6 fetuses had PKD1 variation, and other 14 fetuses had no PKD1/ PKD2-genetic variation. Conclusions:The combination of next-generation sequencing, multiplex ligation-dependent probe amplification, and long-range PCR combined with Sanger sequencing can be helpful for rapid, efficient and accurate genetic diagnosis of ADPKD pedigrees. Point mutations are the most common types in PKD1/PKD2. Fourteen novel variants in PKD1/PKD2 extend its pathogenic variant spectrum and can provide basis for genetic counseling and prenatal diagnosis of ADPKD pedigrees.

BACKGROUND:Whole-exome sequencing (WES) is an effective method in the prenatal setting for identification of the underlying genetic etiology of fetal ultrasound abnormalities. To investigate the diagnostic value of WES in fetuses with ultrasound abnormalities that resulted in fetal demise or pregnancy termination.METHODS:61 deceased fetuses with ultrasound abnormalities and normal copy number variation Sequencing were retrospectively collected. Proband-only or trio-WES were performed on the products of conception.RESULT:Collectively, 28 cases were positive with 39 variants (10 pathogenic, 22 likely pathogenic and 7 variants of uncertain significance) of 18 genes, and the overall diagnostic rate was 45.9% (28/61), of which 39.2% (11/28) were de novo variants. In addition, 21 variants in 11 genes among the positive cases had not been previously reported. The diagnostic yield for definitive findings for trio analysis was 55.9% (19/34) compared to 33.3% (9/27) for singletons. The most common ultrasound abnormalities were skeletal system abnormalities 39.2% (11/28), followed by multiple system abnormalities (17.9%, 5/28) and genitourinary abnormalities (17.9%, 5/28).CONCLUSION:Our results support the use of WES to identify genetic etiologies of ultrasound abnormalities and improve understanding of pathogenic variants. The identification of disease-related variants provided information for subsequent genetic counseling of recurrence risk and management of subsequent pregnancies.

OBJECTIVE:To explore the clinical and genetic features of a child with autosomal dominant mental retardation type 40 (MRD40) due to variant of the CHAMP1 gene.METHODS:Clinical characteristics of the child were analyzed. Genetic testing was carried out by low-depth high-throughput and whole genome copy number variant sequencing (CNV-seq) and whole exome sequencing (WES). A literature review was also carried out for the clinical phenotype and genetic characteristics of patients with MRD40 due to CHAMP1 gene variants.RESULTS:The child, a 11-month-old girl, has presented with intellectual and motor developmental delay. CNV-seq revealed no definite pathogenic variants. WES has detected the presence of a heterozygous c.1908C>G (p.Y636*) variant in the CHAMP1 gene, which was carried by neither parent and predicted to be pathogenic. Literature review has identified 33 additional children from 12 previous reports. All children had presented with developmental delay and mental retardation, and most had dystonia (94.1%), delayed speech and/or walking (85.2%, 82.4%) and ocular abnormalities (79.4%). In total 26 variants of the CHAMP1 gene were detected, with all nonsense variants being of loss-of-function type, located in exon 3, and de novo in origin.CONCLUSION:The heterozygous c.1908C>G (p.Y636*) variant of the CHAMP1 gene probably underlay the WRD40 in this child. Genetic testing should be considered for children featuring global developmental delay, mental retardation, hypertonia and facial dysmorphism.

OBJECTIVE:To assess the diagnostic value of copy number variation sequencing (CNV-seq) in the genetic etiology of fetuses with nasal bone dysplasia (NBD).METHODS:A total of 217 fetuses discovered with NBD from December 2017 to December 2020 were divided into the isolated NBD group and NBD combined with other anomalies group, for which copy number variations (CNVs) were analyzed.RESULTS:A total of 40 fetal abnormalities were detected in 217 cases, with an overall abnormal rate of 18.4%. These included 31 cases with aneuploidies (14.3%, 31/217) and 9 cases with genomic CNVs (4.1%, 9/217). Five cases of trisomy 21 (3.5%, 5/144) and two CNVs cases with unknown clinical significance (1.4%, 2/144) were detected in the isolated group. As for the combined NBD group, 26 aneuploidies (35.6%, 26/73), including 19 cases with trisomy 21, 6 cases with trisomy 18, 1 case with trisomy 13, 5 cases with pathogenic CNVs (6.8%, 5/73), and 2 cases with CNVs of unknown clinical significance (2.7%, 2/73) were detected. A significant difference was detected between the two groups (P < 0.01).CONCLUSION:The detection rate of CNV-seq is high for chromosomal aneuploidies and pathogenic CNVs in fetuses with NBD, particularly in those combined with other ultrasonic abnormalities.

Objective:To explore the genetic basis for a Chinese patient suspected for Canavan disease.Methods:Whole exome sequencing (WES) was carried out for the proband, and candidate variants were verified by Sanger sequencing of the proband, her parents and brother. Prenatal diagnosis was provided to her mother by chorionic villi sampling (CVS) upon her subsequent pregnancy.Results:The proband, a 4-month-old female infant, had manifested drowsiness, hypotonia and apathy. Urine metabolism screening showed elevated N-acetylaspartic acid. Cranial magnetic resonance imaging revealed abnormal myelination and multiple abnormal signals in large brain areas. WES revealed that the proband has harbored compound heterozygous variants of the ASPA gene, namely c. 187A>G (p.Arg63Gly) in exon 1 and c. 634+ 1G>A (P.? ) in exon 4. Sanger sequencing confirmed that the c. 187A>G (p.Arg63Gly) and c. 634+ 1G>A (p.? ) variants were respectively inherited from her mother and father. Her phenotypically normal brother has carried a heterozygous c. 634+ 1G>A (p.? ) variant. Prenatal diagnosis by CVS indicated that the fetus was a heterozygous carrier of the c. 187A>G variant. Conclusion:WES can facilitate the diagnosis of Canavan disease, particularly for those lacking specific phenotypes of the disease. The compound heterozygous variants of the ASPA gene probably underlay the Canavan disease in this patient, and the result has enabled prenatal diagnosis for this family.

Background Phenylketonuria (PKU) is a metabolic disease that can cause severe and irreversible brain damage without treatment. Methods Here we developed a non-invasive prenatal diagnosis (NIPD) technique based on haplotypes via paired-end molecular tags and weighting algorithm and applied it to the NIPD of PKU to evaluate its accuracy and feasibility in the early pregnancy. A custom-designed hybridization probes containing regions in phenylalanine hydroxylase ( PAH ) gene and its 1 Mb flanking region were used for target sequencing on genomic and maternal plasma DNA (7–13 weeks of gestation) to construct the parental haplotypes and the proband’s haplotype. Fetal haplotype was then inferred combined with the parental haplotypes and the proband’s haplotype. The presence of haplotypes linked to both the maternal and paternal mutant alleles indicated affected fetuses. The fetal genotypes were further validated by invasive prenatal diagnosis in a blinded fashion. Results This technique has been successfully applied in twenty-one cases. Six fetuses were diagnosed as patients carrying both of the mutated haplotypes inherited from their parents. Eleven fetuses were carriers of one heterozygous PAH variants, six of which were paternal and five of which were maternal. Four fetuses were absence of pathogenic alleles. All results were consistent with the prenatal diagnosis through amniotic fluid. Conclusions The results showed that our new technique applied to the genotyping of fetuses with high risk for PKU achieves an accurate detection at an early stage of pregnancy with low fetal fraction in cell free DNA.

OBJECTIVE:To describe the clinical and genetic characteristics of a child with 14q12q13.1 deletion involving the FOXG1 gene.METHODS:Clinical manifestation of the child was analyzed. Peripheral blood sample of the patient was subjected to chromosomal karyotyping and single nucleotide polymorphism array (SNP-array) analysis.RESULTS:The male infant has developed feeding difficulty, poor sucking, lower limb tremor, and frontal bruising 8 days after birth. Magnetic resonance imaging revealed significant enlargement of bilateral ventricles and corpus callosum dysplasia. Chromosomal analysis revealed a karyotype of 46,XY,del(14)(q12q13.1), and SNP-array confirmed that there was a 9.6 Mb deletion in 14q11.2q13.1, which encompassed the FOXG1 gene.CONCLUSION:For patients with brain development abnormalities, dyskinesia, cognitive impairment, speech disorder and other manifestations, copy number variation of the FOXG1 gene should be excluded. SNP-array should be carried out as early as possible to attain the diagnosis.

目的 分析了3334例符合河南省东北片区(郑州大学第一附属医院片区)民生实事免费产前诊断胎儿染色体核型结果,探讨不同产前指征的异常率及胎儿染色体异常核型的类型、发生频率及相关因素,为临床遗传咨询提供依据.方法 收集2019年7月至2020年2月符合河南省民生实事产前诊断指征的3343例孕妇进行免费的羊膜腔穿刺术及羊水细胞培养,并对分析的3334例胎儿染色体核型结果进行分析统计.结果 共穿刺培养3343例,培养失败9例,获得染色体核型结果3334例,成功率为99.73％.3334例孕妇中共检出染色体异常121例,总异常率3.63％.其中数目异常48例(1.44％)(包括21三体33例,18三体5例,嵌合型13三体1例),结构异常39例(1.17％),多态性36例(1.08％).按不同产前诊断指征分组统计,异常发生率分别为:唐筛高风险组3.06％(69/2255),高龄组3.79％(37/977),胎儿父母为平衡易位或倒位组23.21％(13/56)和非整倍体患儿出生史组4.35％(2/46).结论 河南省民生实事项目的 开展对这4种产前指征的孕妇进行免费羊水染色体核型分析,提高了孕妇及家属产前诊断的意识,并切实地降低了缺陷患儿的出生,对预防出生缺陷具有重要意义.

With an incidence of 1/800 - 1/600, Down syndrome (DS) is the most common chromosomal disorder in humans. Whilst most DS patients has trisomy 21, a small proportion may carry translocations or mosaicisms involving chromosome 21. The main characteristics of DS include mental retardation, peculiar facies, growth retardation, congenital heart disease, duodenal stenosis, Alzheimer's disease, leukemia, and immunodeficiency, which may be due to increased dosage of critical genes. Recent studies also showed that epigenetic changes may also occur in DS. For research on patients with DS or other trisomies have been restricted by ethical considerations, and commonly used mouse models cannot fully replicate the characteristic features of DS, pluripotent stem cells induced from fetal samples or biopsy tissues from DS patients may generate models with the same genetic content, which may provide idea materials for studying the pathogenesis of DS and customized cell and/or gene therapies.

Serine protease inhibitors (SPI) are a superfamily of the proteins able to suppress serine protease activity, and may exert the major biological function in complement activation, inflammation, and fibrinolysis. A SPI was identified from Trichinella spiralis adult worms (AW) by immunoproteomics with early infection sera. The aim of this study was to investigate the protective immune elicited by TsSPI. The complete TsSPI cDNA sequence was cloned into pQE-80 L and then expressed in Escherichia coli BL21. The rTsSPI was purified and its antigenicity was determined by Western blotting analysis. By using anti-rTsSPI serum the native TsSPI was identified in somatic and ES proteins from muscle larvae (ML). The results of qPCR and immunofluorescence assay (IFA) revealed that the expression of the TsSPI gene was observed throughout all developmental stages of T. spiralis (ML, intestinal infective larvale, 3- and 6-days AW, and newborn larvae, NBL), located principally in cuticles, stichosome, and embryos of this parasitic nematode. Vaccination of mice with rTsSPI triggered high level of anti-TsSPI IgG response, and showed a 62.2 and 57.25% worm burden reduction in the recovery of intestinal AW at 6 days post-infection (dpi) and ML at 35 dpi, respectively. The TsSPI might be a novel potential target for anti-Trichinella vaccine.

•Surface proteins of T. spiralis ML and IIL were analyzed.•Forty one proteins were common to both larvae, 62 biased ML and 113 biased IIL.•Proteins of nitrogen compound metabolic process were highly expressed in IIL.•qPCR results confirmed that 4 of 6 IIL genes selected were up-regulated.•These proteins might be invasion-related proteins and vaccine candidate antigens.