RESUMEN
Hereditary tyrosinemia type 1 (HT1) is a severe human autosomal recessive disorder caused by the deficiency of fumarylacetoacetate hydroxylase (FAH), an enzyme catalyzing the last step in the tyrosine degradation pathway. Lack of FAH causes accumulation of toxic metabolites (fumarylacetoacetate and succinylacetone) in blood and tissues, ultimately resulting in severe liver and kidney damage with onset that ranges from infancy to adolescence. This tissue damage is lethal but can be controlled by administration of 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC), which inhibits tyrosine catabolism upstream of the generation of fumarylacetoacetate and succinylacetone. Notably, in animals lacking FAH, transient withdrawal of NTBC can be used to induce liver damage and a concomitant regenerative response that stimulates the growth of healthy hepatocytes. Among other things, this model has raised tremendous interest for the in vivo expansion of human primary hepatocytes inside these animals and for exploring experimental gene therapy and cell-based therapies. Here, we report the generation of FAH knock-out rabbits via pronuclear stage embryo microinjection of transcription activator-like effector nucleases. FAH-/- rabbits exhibit phenotypic features of HT1 including liver and kidney abnormalities but additionally develop frequent ocular manifestations likely caused by local accumulation of tyrosine upon NTBC administration. We also show that allogeneic transplantation of wild-type rabbit primary hepatocytes into FAH-/- rabbits enables highly efficient liver repopulation and prevents liver insufficiency and death. Because of significant advantages over rodents and their ease of breeding, maintenance, and manipulation compared with larger animals including pigs, FAH-/- rabbits are an attractive alternative for modeling the consequences of HT1.
Asunto(s)
Hidrolasas/genética , Tirosinemias/genética , Animales , Modelos Animales de Enfermedad , Femenino , Técnicas de Inactivación de Genes , Hepatocitos/trasplante , Humanos , Hidrolasas/metabolismo , Riñón/metabolismo , Riñón/patología , Hígado/metabolismo , Hígado/patología , Fallo Hepático/etiología , Fallo Hepático/metabolismo , Fallo Hepático/patología , Fallo Hepático/terapia , Masculino , Conejos , Tirosinemias/complicaciones , Tirosinemias/metabolismo , Tirosinemias/patologíaRESUMEN
Aldosterone is a mineralocorticoid steroid hormone, the measurement of which in the clinical laboratory is principally performed for the investigation of primary hyperaldosteronism. Primary hyperaldosteronism is a specifically treatable and potentially curable form of hypertension, which typically presents as drug-resistant hypertension and, in up to 37% of cases, hypokalemia. Accurate measurement of aldosterone concentration is essential for correct diagnosis. The serum concentrations of aldosterone are in the picomolar range and therefore sensitive aldosterone assays are required. With the advancement in instrumentation of LC-MS/MS, the picomolar range of aldosterone can be easily measured by the newer models, but for those with a less sensitive instrument, special technique for sample preparation to enhance assay sensitivity is required. This work described the use of charge-tagging for the picomolar measurement of serum aldosterone in a less sensitive LC-MS/MS instrument. The assay was linear up to 3000 pmol/L with lower limit of quantitation at 80 pmol/L. The mean relative recovery was 96.5% with a range of 89.3-101.6% for aqueous calibrators and the mean relative recovery was 94.8% with a range of 87.5-101.4% for serum calibrators. Intra-assay CVs range from 8.2% to 11.3%, and inter-assay CVs ranged from 8.5% to 13.5% at concentration range from 229 to 1720 pmol/L. The LC-MS/MS method compared well (y = 1.04x + 8.97) with the in-use radioimmunoassay method. There was no significant difference found (p = 0.7135) between results determined by LC-MS/MS and radioimmunoassay method.
Asunto(s)
Aldosterona/sangre , Hiperaldosteronismo/sangre , Espectrometría de Masas en Tándem/métodos , Adulto , Anciano , Calibración , Cromatografía Liquida/métodos , Femenino , Humanos , Hiperaldosteronismo/diagnóstico , Límite de Detección , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Adulto JovenRESUMEN
Influenza-associated encephalopathy (IAE) is a potentially fatal neurological complication of influenza infection usually in the presence of high and persistent fever. Thermolabile carnitine palmitoyltransferase II enzyme (CPT-II) predisposes IAE, so far only described in Japanese. As the genetic origins of Japanese and Chinese are alike, similar genetic risk factors in CPT-II are expected. We report the first two unrelated Chinese patients of thermolabile CPT-II variants that underlain the persistent high fever-triggered viral infection-associated encephalopathy, multi-organ failure and death. Elevated (C16:0+C18:1)/C2 acylcarnitines ratio and the CPT2 susceptibility variant allele [p.Phe352Cys; p.Val368Ile] were detected. The asymptomatic family members of one patient also had abnormal long-chain acylcarnitines. In our experience of biochemical genetics, the elevated (C16:0+C18:1)/C2 acylcarnitines ratio is unusual and specific for thermolabile CPT-II variants. Allele frequency of [p.Phe352Cys; p.Val368Ile] among Hong Kong Chinese was 0.104, similar to Japanese data, and [p.Phe352Cys] has not been reported in Caucasians. This may explain the Asian-specific phenomenon of thermolabile CPT-II-associated IAE. We successfully demonstrated the thermolabile CPT-II variants in patients with viral infection-associated encephalopathy in another Asian population outside Japanese. The condition is likely under-recognized. With our first cases, it is envisaged that more cases will be diagnosed in subsequent years. The exact pathogenic mechanism of how other factors interplay with thermolabile CPT-II variants and high fever leading to IAE, is yet to be elucidated. Fasting and decreased intake during illness may aggravate the disease. Further studies including high risk and neonatal screening are warranted to investigate its expressivity, penetrance and temperature-dependent behaviors in thermolabile CPT-II carriers. This may lead to discovery of the therapeutic golden window by aggressive antipyretics and L-carnitine administration in avoiding the high mortality and morbidity of IAE.
Asunto(s)
Carnitina O-Palmitoiltransferasa/metabolismo , Encefalitis Viral/enzimología , Gripe Humana/complicaciones , Sustitución de Aminoácidos , Secuencia de Bases , Carnitina/análogos & derivados , Carnitina/metabolismo , Carnitina O-Palmitoiltransferasa/genética , Preescolar , Análisis Mutacional de ADN , Encefalitis Viral/complicaciones , Encefalitis Viral/genética , Estabilidad de Enzimas , Salud de la Familia , Resultado Fatal , Femenino , Predisposición Genética a la Enfermedad/genética , Genotipo , Humanos , Masculino , Datos de Secuencia Molecular , Mutación , Linaje , Factores de Riesgo , TemperaturaRESUMEN
Metformin is a widely used antidiabetic agent that is generally considered safe. However, metformin-associated lactic acidosis (MALA), though not common, occurs from time to time and results in significantly high mortality. A series of 23 MALA cases in a local major hospital in Hong Kong is reported in this article to demonstrate the epidemiological data, risk factors, clinical features as well as the clinical outcomes for better understanding of this disease entity. It is the first MALA case series in which plasma metformin levels were assessed. However, the results show that plasma metformin levels in MALA bear no diagnostic and prognostic values. Risk factors of mortality were identified as shock and high plasma lactate levels. The majority of patients were found to have significantly raised creatinine versus a normal baseline value before the acute illness. Concomitant illnesses taking place alongside MALA were common. With a high utility rate of renal replacement therapy (82.6%) in the study group, the mortality rate was 30.4%.
Asunto(s)
Acidosis Láctica/inducido químicamente , Diabetes Mellitus Tipo 2/complicaciones , Hipoglucemiantes/efectos adversos , Metformina/efectos adversos , Acidosis Láctica/sangre , Acidosis Láctica/epidemiología , Adulto , Anciano , Anciano de 80 o más Años , Amilasas/sangre , Pueblo Asiatico , China/epidemiología , Creatinina/sangre , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/epidemiología , Diabetes Mellitus Tipo 2/terapia , Sobredosis de Droga , Femenino , Cardiopatías/epidemiología , Humanos , Hipertensión/epidemiología , Hipoglucemiantes/sangre , Enfermedades Renales/epidemiología , Masculino , Metformina/sangre , Persona de Mediana Edad , Terapia de Reemplazo Renal , Factores de Riesgo , Accidente Cerebrovascular/epidemiologíaRESUMEN
Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was applied to the direct analysis of melamine cyanurate (MC). The three commonly used MALDI matrixes, namely, alpha-cyano-4-hydroxycinnamic acid (CHCA), sinapinic acid (SA), and 2,5-dihydroxybenzoic acid (DHB), were able to desorb/ionize melamine from MC upon N(2) laser irradiation, with CHCA showing the highest detection sensitivity in the positive mode. Only DHB and SA were able to desorb/ionize cyanuric acid from MC in the negative mode but with remarkably lower sensitivity. The method is able to detect melamine unambiguously from a small amount of MC (down to 12.5 microg) spiked into urine and was successfully applied for the rapid and sensitive detection of melamine in urine stones/residues of the samples collected from patients clinically confirmed of having kidney stones associated with the consumption of melamine-tainted food products. The urine matrix resulted in interfering ion peaks and suppressed the ion intensity of melamine, while a cleanup process consisting of simply washing with water eliminated such interference and enhanced the ion intensity. The merit of the method is simplicity in sample preparation. The analytical time of the method for high-throughput analysis from the time of sample treatment to analysis is less than 7 minutes per sample, with sensitive detection of the presence of melamine in the urine stones/residues of the patient samples.
Asunto(s)
Triazinas/orina , Niño , Contaminación de Alimentos , Humanos , Cálculos Renales/etiología , Cálculos Renales/orina , Sensibilidad y Especificidad , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Triazinas/análisis , Triazinas/químicaRESUMEN
Cough mixture abuse has been reported to cause severe folate deficiency and neurological defects. We carried out a prospective case-controlled survey to confirm this association and define the incidence and severity of the problem. A total of 57 cough mixture abusers and 47 other substance abusers (controls) were studied. When compared with controls, cough mixture abusers had a high incidence of low folate levels that could only be detected by screening.
Asunto(s)
Antitusígenos/efectos adversos , Codeína/efectos adversos , Dextrometorfano/efectos adversos , Deficiencia de Ácido Fólico/inducido químicamente , Trastornos Relacionados con Sustancias/complicaciones , Adulto , Estudios de Casos y Controles , Recolección de Datos , Caries Dental/inducido químicamente , Índices de Eritrocitos , Femenino , Deficiencia de Ácido Fólico/sangre , Deficiencia de Ácido Fólico/epidemiología , Hemoglobinas/análisis , Hong Kong/epidemiología , Humanos , Incidencia , Masculino , Tamizaje Masivo , Enfermedades del Sistema Nervioso/inducido químicamente , Recuento de Plaquetas , Estudios Prospectivos , Trastornos Relacionados con Sustancias/sangre , Trastornos Relacionados con Sustancias/epidemiología , Deficiencia de Vitamina B 12/sangre , Deficiencia de Vitamina B 12/inducido químicamente , Deficiencia de Vitamina B 12/epidemiologíaRESUMEN
The conventional method for the determination of mercury in clinical samples is cold vapor atomic absorption spectrometry. Sample digestion or pretreatment require large sample volume and long sample preparation time. The inductively coupled plasma mass spectrometry (ICP-MS) method developed in this study requires only 100 microL of sample with practically no preparation, except for dilution with diluent. Significant savings in sample volumes, reagents, technician time, and analysis time are realized. Among different types of diluents, the one containing acid, tert-butanol, and potassium dichromate gave the best results to remove the mercury memory effect. The interassay precisions for whole blood and urine were < 5% and < 8%, respectively, and the intra-assay precisions were < 3% and < 7%, respectively. The lower limits of detection were 0.13, 0.17, and 0.26 microg/L for aqueous standard, urine, and whole blood, respectively. The developed ICP-MS method correlated well with the atomic absorption method and can offer an alternative to the atomic absorption method for mercury analysis with less sample volume requirement as well as shorter analysis time.
Asunto(s)
Contaminantes Ambientales/sangre , Contaminantes Ambientales/orina , Mercurio/sangre , Mercurio/orina , Humanos , Ácido Clorhídrico , Indicadores y Reactivos , Espectrometría de Masas , Dicromato de Potasio , Alcohol terc-ButílicoAsunto(s)
Enfermedades Renales/genética , Síndrome Metabólico/genética , Síndrome Metabólico/metabolismo , Transportadores de Anión Orgánico/genética , Proteínas de Transporte de Catión Orgánico/genética , Ácido Úrico/metabolismo , Adulto , Anciano , Creatinina/sangre , Análisis Mutacional de ADN , Diabetes Mellitus Tipo 2/genética , Familia , Femenino , Humanos , Enfermedades Renales/metabolismo , Pruebas de Función Renal , Masculino , Mutación/genéticaRESUMEN
BACKGROUND: Carnitine palmitoyltransferase II (CPT II) deficiency is one of the most common disorders of oxidative fatty acid metabolism. In this disorder, long-chain acylcarnitines cannot be converted to acyl CoA and there is impairment of ß-oxidation of fatty acids. RESULTS: In the 3 distinct clinical subtypes of CPT II deficiency, adult onset myopathic form shows mild clinical manifestations, characterized by recurrent rhabdomyolysis after intense physical stress. In this study, we report a case with adult myopathic form of CPT II deficiency presenting recurrent exercise-induced myoglobinuria. CONCLUSION: The acylcarnitine profile showed characteristic CPTII deficiency profile and sequencing of the CPT2 gene showed 2 novel missense mutations p. H369Q and p G497S.
Asunto(s)
Carnitina O-Palmitoiltransferasa/deficiencia , Carnitina O-Palmitoiltransferasa/genética , Errores Innatos del Metabolismo/genética , Mutación Missense , Mioglobinuria/genética , Adulto , Pueblo Asiatico , Secuencia de Bases , Humanos , Masculino , Errores Innatos del Metabolismo/complicaciones , Errores Innatos del Metabolismo/diagnóstico , Datos de Secuencia Molecular , Mioglobinuria/complicaciones , Mioglobinuria/diagnóstico , Esfuerzo FísicoRESUMEN
OBJECTIVES: Tyrosinemia type I is an autosomal recessive disorder in tyrosine metabolism. In areas without expanded newborn screening, patients present with acute hepatorenal failure in early infancy. Diagnosis can be elusive when clinical presentation is non-specific and biochemical abnormalities are masked by secondary changes. This is the first Hong Kong Chinese report. DESIGN AND METHODS: A two-month-old Chinese male infant with unremarkable antenatal and postnatal history presented with progressive abdominal distension for three days. He suffered from end-stage liver failure, hypoglycemia and hepatic encephalopathy. Diagnostic work-up was complicated starting from rule-out sepsis, intestinal obstruction, volvulus, peritonitis, septic ileus, poisoning to metabolic diseases. Clinical, biochemical and genetic data was described. RESULTS: The patient showed increases in multiple plasma amino acids including tyrosine, phenylalanine and methionine, and hyper-excretions of 4-hydroxyphenyl-acetate, -pyruvate, and -lactate, as well as N-acetyltyrosine which could be seen in liver failure due to both tyrosinemia type I and non-metabolic conditions. Because of the volatile nature, succinylacetone was almost undetectable. The diagnosis was confirmed by genetic analysis of FAH with two novel mutations, viz. NM_000137.2:c.1063-1G>A and NM_000137.2:c.1035_1037del. Living-related liver transplantation was done. However, the patient still suffered many complications after the severe metabolic insult with hypoxic ischemic encephalopathy, cerebral atrophy, global developmental delay and cortical visual impairment. CONCLUSIONS: Because of the lack of expanded newborn screening in Hong Kong, this child unfortunately presented in the most severe form of tyrosinemia type I. Expanded newborn screening can save life and reduce the burden of diagnostic complexity. This illustrates the need for expanded newborn screening in Hong Kong.
Asunto(s)
Hidrolasas/genética , Mutación , Tirosinemias/diagnóstico , Tirosinemias/genética , Hong Kong , Humanos , Lactante , Recién Nacido , Fallo Hepático/etiología , Fallo Hepático/genética , Masculino , Tamizaje Neonatal , Tirosinemias/etiologíaRESUMEN
Very long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is one of the most common fatty acid oxidation defects that cause sudden unexpected deaths in infants. The death attributed to VLCAD deficiency can be prevented by early diagnosis with expanded newborn screening using tandem mass spectrometry. A favorable outcome can be achieved with early diagnosis and prompt treatment. However, such newborn screening has not yet been available in Hong Kong. We report a 2-month-old boy who succumbed 5 hours after admission with the diagnosis of VLCAD deficiency confirmed by genetic analysis performed after death. The patient was compound heterozygous for a novel splicing mutation ACADVL NM_000018.2:c.277+2T>G; NC_000017.10:g.7123997T>G and a known disease-causing mutation ACADVL NM_000018.2:c.388_390del; NP_000009.1: p.Glu130del. Family screening was performed for at-risk siblings. The rapid downhill course of the patient clearly illustrates the need of newborn screening for early diagnosis. Our patient was asymptomatic before metabolic decompensation. However, once metabolic decompensation occurred, rapid deterioration and death followed, which obviated the opportunity to diagnose and treat. The only way to save these patients' lives and improve their outcome is early diagnosis and appropriate treatment. Therefore, we strongly urge the implementation of newborn screening using tandem mass spectrometry for VLCAD deficiency and other highly treatable inborn errors of metabolism in Hong Kong.
Asunto(s)
Acil-CoA Deshidrogenasa de Cadena Larga/genética , Errores Innatos del Metabolismo Lipídico/diagnóstico , Enfermedades Mitocondriales/diagnóstico , Enfermedades Musculares/diagnóstico , Tamizaje Neonatal , Acil-CoA Deshidrogenasa de Cadena Larga/sangre , Acil-CoA Deshidrogenasa de Cadena Larga/deficiencia , Síndromes Congénitos de Insuficiencia de la Médula Ósea , Resultado Fatal , Heterocigoto , Hong Kong , Humanos , Lactante , Recién Nacido , Errores Innatos del Metabolismo Lipídico/genética , Errores Innatos del Metabolismo Lipídico/prevención & control , Masculino , Enfermedades Mitocondriales/genética , Enfermedades Mitocondriales/prevención & control , Enfermedades Musculares/genética , Enfermedades Musculares/prevención & control , Mutación , Empalme del ARN/genética , Análisis de Secuencia de ADN , Hermanos , Espectrometría de Masas en TándemRESUMEN
BACKGROUND: The diagnosis of aromatic L-amino acid decarboxylase (AADC) deficiency, one of the pediatric neurotransmitter disorders, is classically made with plasma enzyme level or cerebrospinal fluid (CSF) neurotransmitter profile, while both are technically demanding and the latter requires the invasive lumbar puncture. So far less than 100 cases have been reported worldwide with 20% from Taiwan. It was postulated that the condition might have been under-diagnosed among Chinese populations and a non-invasive screening tool should be developed in areas with high prevalence. METHODS: Urine metabolic profiles performed by gas chromatography-mass spectrometry (GC-MS) in a 31-month period were retrospectively reviewed: those with vanilmandelic acid concentration lower than one percentile plus the presence of 3-o-methyldopa were defined as positive and the patients were further evaluated. RESULTS: Among 1046 metabolic profiles (from 845 patients) reviewed, 3 profiles from 2 patients were screened positive: both cases had compatible CSF neurotransmitter profiles and the diagnosis was further confirmed by genetic analysis of DDC gene. 13 negative urinary metabolic profiles from 7 patients who had CSF neurotransmitters analyzed were identified as controls: all 7 CSF neurotransmitter profiles were not compatible for AADC deficiency. CONCLUSIONS: The GC-MS-based urine metabolic profiling was shown to be a satisfactory screening tool for AADC deficiency. Further confirmation can be performed by mutation analysis in the DDC gene, thus avoiding risks of lumbar puncture. We advocate all ethnic Chinese patients presenting with dystonia have their urine organic acids analyzed before proceeding to CSF neurotransmitters analysis.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/orina , Descarboxilasas de Aminoácido-L-Aromático/deficiencia , Descarboxilasas de Aminoácido-L-Aromático/orina , Cromatografía de Gases y Espectrometría de Masas , Hong Kong/epidemiología , Humanos , Lactante , Masculino , Proyectos Piloto , PrevalenciaRESUMEN
In areas without expanded newborn screening, instead of presenting neonatally, patients with arginase deficiency typically present with spastic paraplegia in early childhood. Diagnosis of this rare neurometabolic disease poses the first challenge because it is often misdiagnosed as cerebral palsy during initial stages. We describe arginase deficiency in a 20-year-old woman with spastic paraplegia, progressive dystonia, dementia, peripheral neuropathy, epilepsy, liver cirrhosis, and non-B/non-C hepatocellular carcinoma. A novel homozygous mutation NM_000045.2 (ARG1):c.673del (p.Arg225GlyfsX5) was detected. We suggest that all children presenting with progressive neurodegeneration or spastic paraplegia in the absence of risk factors for cerebral palsy should be screened for inborn errors of metabolism, including arginase deficiency. For monitoring urea cycle defects, noninvasive imaging screening for liver fibrosis and hepatocellular carcinoma can help ensure early detection, with potential treatment implications.
Asunto(s)
Arginasa/genética , Hiperargininemia/genética , Eliminación de Secuencia , Anticonvulsivantes/uso terapéutico , Arginasa/fisiología , Secuencia de Bases , Carcinoma Hepatocelular/diagnóstico por imagen , Carcinoma Hepatocelular/etiología , Parálisis Cerebral/diagnóstico , Codón sin Sentido , Terapia Combinada , Contraindicaciones , Diagnóstico Tardío , Demencia/etiología , Errores Diagnósticos , Progresión de la Enfermedad , Epilepsia/tratamiento farmacológico , Epilepsia/etiología , Resultado Fatal , Femenino , Humanos , Hiperargininemia/diagnóstico , Hiperargininemia/dietoterapia , Hiperargininemia/tratamiento farmacológico , Hígado/enzimología , Cirrosis Hepática/diagnóstico por imagen , Cirrosis Hepática/etiología , Neoplasias Hepáticas/diagnóstico por imagen , Neoplasias Hepáticas/etiología , Datos de Secuencia Molecular , Cuidados Paliativos , Fenotipo , Radiografía , Benzoato de Sodio/uso terapéutico , Ultrasonografía , Ácido Valproico , Adulto JovenRESUMEN
BACKGROUND: Data of classical inborn errors of metabolism (IEM) of amino acids, organic acids and fatty acid oxidation are largely lacking in Hong Kong, where mass spectrometry-based expanded newborn screening for IEM has not been initiated. The current study aimed to evaluate the approximate incidence, spectrum and other characteristics of classical IEM in Hong Kong, which would be important in developing an expanded newborn screening program for the local area. METHODS: The laboratory records of plasma amino acids, plasma acylcarnitines and urine organic acids analyses from year 2005 to 2009 inclusive in three regional chemical pathology laboratories providing biochemical and genetic diagnostic services for IEM were retrospectively reviewed. RESULTS: Among the cohort, 43 patients were diagnosed of IEM, including 30 cases (69%) of amino acidemias (predominantly citrin deficiency, hyperphenylalaninemia due to 6-pyruvoyl-tetrahydropterin synthase deficiency and tyrosinemia type I), 5 cases (12%) of organic acidemias (predominantly holocarboxylase synthetase deficiency) and 8 cases (19%) of fatty acid oxidation defects (predominantly carnitine-acylcarnitine translocase deficiency). The incidence of classical IEM in Hong Kong was roughly estimated to be at least 1 case per 4122 lives births, or 0.243 cases per 1000 live births. This incidence is similar to those reported worldwide, including the mainland of China. The estimated incidence of hyperphenylalaninemia was 1 in 29 542 live births. CONCLUSIONS: Our data indicate that it is indisputable for the introduction of expanded newborn screening program in Hong Kong. Since Hong Kong is a metropolitan city, a comprehensive expanded newborn screening program and referral system should be available to serve the neonates born in the area.
Asunto(s)
Errores Innatos del Metabolismo/diagnóstico , Tamizaje Neonatal/métodos , Ácidos/orina , Aminoácidos/sangre , Carnitina/análogos & derivados , Carnitina/sangre , Hong Kong/epidemiología , Humanos , Recién Nacido , Errores Innatos del Metabolismo/sangre , Errores Innatos del Metabolismo/epidemiología , Errores Innatos del Metabolismo/orina , Espectrometría de Masas en TándemRESUMEN
Glutaric aciduria type II, or multiple acyl-CoA dehydrogenase deficiency, is a rare metabolic disorder inherited in an autosomal recessive manner. The condition can be caused by mutations in at least 3 genes, including ETFA, ETFB, and ETFDH. When this potentially lethal disorder is known for its clinical and biochemical heterogeneity, mutation analysis will be an invaluable part of diagnosis. We here described a Chinese adolescent boy who enjoyed good health earlier and presented at the age of 14 years with severe vomiting. His condition deteriorated rapidly and he succumbed shortly after. With a travel history before presentation and the late age of onset, diagnosis was particularly difficult. Findings in perimortem biochemical investigations and postmortem autopsy were guiding but not diagnostic. The diagnosis of glutaric aciduria type II was finally confirmed by mutation analysis performed by direct sequencing on genomic DNA from peripheral blood, which identified 2 different unreported missense mutations, c.502G>T (p.V168F) and c.786A>G (p.Q262R), in ETFA. The father and the mother were found to be heterozygous for the 2 mutations in ETFA respectively. Subsequent molecular family screening also ruled out the disease in his elder sister, who had a history of convulsion and a suspicious plasma acylcarnitine profile, and freed her from life-long supplementation. The case showed that molecular autopsies should be part of routine postmortem examination of unexplained sudden death in all age groups and DNA-friendly samples should be routinely collected and archived. In the era of personalized medicine with the power of modern genetics, molecular diagnosis should be obtained for heterogeneous diseases with different genetic defects but sharing similar clinical and/or biochemical phenotypes.
Asunto(s)
Diagnóstico , Pruebas Genéticas , Deficiencia Múltiple de Acil Coenzima A Deshidrogenasa/diagnóstico , Deficiencia Múltiple de Acil Coenzima A Deshidrogenasa/patología , Patología Molecular/métodos , Adolescente , Pueblo Asiatico , China , Resultado Fatal , Humanos , MasculinoRESUMEN
BACKGROUND: Trace element determination in laboratory medicine is widely carried out by atomic absorption or emission spectroscopy. In the last decade, there has been a rapid growth in the use of inductively coupled plasma-mass spectrometry because of its strong detection power, and the possibility of multi-elements analysis in a single run. METHODS: Having the advantages of smaller sample volume and better detection limit, we developed a method for the simultaneous determinations of six trace elements by using 100 microL serum, and the assay can be accomplished within 3 min. RESULTS: The method developed gave recovery of the six elements ranging from 97% to 117%. The method covered a wide dynamic range with manganese in the range of nmol/L, while magnesium was in the range of mmol/L. The detection limits were 0.001 mmol/L, 0.05 micromol/L, 2.0 nmol/L, 0.2 micromol/L, 0.05 micromol/L, and 0.01 micromol/L for magnesium, aluminium, manganese, copper, zinc, and selenium, respectively. All the six elements had intra-assay imprecision of less than 5%, and inter-assay imprecision of less than 8%. CONCLUSIONS: This fast and robust method is suitable for use in the clinical laboratory where short turnaround time is needed for managing patients with trace element deficiency or toxicity.