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Objective:To investigate the accuracy, effectiveness and feasibility of MassARRAY genotyping assay in the diagnoses of neonatal genetic metabolic diseases.Methods:This is a retrospective study. From December 2016 to January 2020, newborns were screened by tandem mass spectrometry at the Zhejiang Newborn Screening Center, among which the data of 7 922 suspected positive cases of genetic metabolic diseases were collected. These patients were then tested for the common variants of 27 genetic metabolic diseases by MassARRAY genotyping assay, along with further testing using Sanger or next-generation sequencing used to verify and/or further search for potential variants.Results:A total of 1 408 cases were tested with MassARRAY. Among these, 307 cases were confirmed with certain genetic metabolic diseases. The detection rate of hyperphenylalaninemia was the highest, followed by primary carnitine deficiency, short acyl-coA dehydrogenase deficiency and methylmalonic acidemia. With these cases, the consistency of Sanger sequencing and MassARRAY was 100% (307/307). Another 287 cases were identified as carriers by MassARRAY with a 49.1% (141/287) consistency in reference to Sanger sequencing, mainly involving SLC22A5 and MCCC1 genes. Meanwhile, 50.8% (146/287) of these cases were found to have another variant mainly involving PAH, PTS and ACADS genes. The remaining 814 cases have no variants; 158 cases out of these patients have continuously abnormal amino acids, acyl carnitines, urine organic acid and/or other biochemical indices, and were tested by next-generation sequencing, among which 38% (60/158) were detected with two variants. In this study, a total of 513 patients with genetic metabolic disease were diagnosed, and the detection rate of MassARRAY was 59.8% (307/513). Conclusions:MassARRAY genotyping assay can be used as an early molecular screening method for neonatal genetic metabolic diseases. The detection rate is particularly high in diseases with a high concentration of hotspot variants, such as hyperphenylalaninemia and primary carnitine deficiency. The future application value of MassARRAY should be further improved by continuously optimizing its ability to identify new disease genes and potential variable sites.
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Alternative splicing of pre-messenger RNA (pre-mRNA) is a crucial mechanism for the diversity of the human transcriptome and proteome. Alternative splicing is a complex gene regulation process. Whole-transcriptome analysis shows that 95% of human exonic genes are alternatively spliced, involving various cis-acting elements and trans-acting factors. Any changes in any component or step may cause erroneous splicing events and lead to the occurrence of various related diseases. In addition to gene replacement therapy that directly changes the splicing results, RNA splicing modification is expected to become a new therapeutic strategy to alleviate or treat diseases by targeting and correcting abnormal pre-mRNA splicing. Splicing modification tools currently developed including RNA trans-splicing, antisense oligonucleotides, small interfering RNA, and small molecule drugs can correct abnormal splicing through different ways. This article reviews the resent progress of epigenetic regulation of pre-mRNA alternative splicing in recent years, and discusses the occurrence and regulation of alternative splicing, the types of diseases caused by related splicing defects, and the current-used tools for targeting and altering splicing. The importance of splicing modification strategies in the future treatment of human diseases is envisioned.
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With the maturity of the technique of adult liver transplantation, pediatric liver transplantation has been gradually emerging in major liver transplantation centers throughout China. Pediatric liver transplantation differs from adult liver transplantation in terms of recipient selection, technical details, perioperative management, postoperative treatment and follow-up, etc. Multidisciplinary cooperation is required to continuously improve the clinical efficacy of pediatric liver transplantation. In this article, we reviewed the significance of multidisciplinary cooperation in achieving the optimal clinical efficacy of pediatric liver transplantation, in respect to the recipient selection and extrahepatic organ function evaluation, mastering the technical key points of different types, improving the quality of postoperative follow-up, and formulating clinical diagnosis and treatment strategies, etc.
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Objective@#To establish a method of calibration and performance verification of the GCMS-QP 2010 Ultra gas chromatography-mass spectrometer for urine organic acid detection after annual maintenance, so as to ensure that the performance of the detection system can continuously meet the detection needs. @*Methods@#According to the requirements of the equipment manufacturer and the Calibration Specifications for Benchtop Gas Chromatography-Mass Spectrometer, the gas chromatography-mass spectrometer was calibrated after annual maintenance. According to the CNAS-CL02-A003 Guidance on the Application of Accreditation Criteria for the Medical Laboratory Quality and Competence in the Field of Clinical Chemistry, the analytical performance of the maintained gas chromatography-mass spectrometer was validated. @*Results@#The calibration results of quality range, quality accuracy, resolution, signal-to-noise ratio and repeatability were all met the requirements. The detection results of internal quality control materials were under control, and the results of retained samples kept unchanged. @*Conclusion@#The calibration and performance verification method of the gas chromatography-mass spectrometer for urine organic acid detection after annual maintenance is established successfully, which is of importance for providing accurate and reliable results of urine organic acid.