RESUMEN
BACKGROUND AND OBJECTIVE: Hypothyroidism and autoimmune thyroiditis are more prevalent than previously considered in women during pregnancy and the postpartum, and are associated with adverse effects on the mother and her fetus. We determined the efficacy and accuracy of screening women for primary hypothyroidism and autoimmune thyroiditis by testing TSH and two thyroid antibodies (TAb): thyroperoxidase antibodies (TPOAb) and thyroglobulin antibodies (TgAb), in eluates of filter paper specimens collected during early pregnancy and the postpartum. METHODS: We enrolled 494 first-trimester pregnant women with no exclusion criteria into a prospective study to detect primary hypothyroidism and autoimmune thyroiditis. Finger stick blood was applied to filter paper, dried in room air, eluted, and promptly tested for TSH and TAb. A total of 178 of the pregnant women (36%) were tested in the early postpartum. Women with abnormal results had confirmatory serum tests. RESULTS: It was found that 91 pregnant women (18.4%) and 43 postpartum women (24.2%) had abnormal TSH values (>4.0 mU/L) and/or positive TAb; 140 pregnant women (28.3%) had TSH values >2.5 mU/L. All subjects with TSH values >4.0 mU/L tested positive for TAb. Eighteen women (3.6%) who tested normal during pregnancy tested abnormal in the postpartum. CONCLUSIONS: This study confirms that TSH and TPOAb measured in eluates of blood-spotted filter paper specimens are excellent screening tests to detect primary hypothyroidism and autoimmune thyroiditis in pregnant and postpartum women. Results are very comparable to serum data in this population published in the literature.
Asunto(s)
Pruebas con Sangre Seca/métodos , Hipotiroidismo/diagnóstico , Tiroiditis Autoinmune/diagnóstico , Tirotropina/sangre , Adolescente , Adulto , Recolección de Muestras de Sangre/métodos , Femenino , Humanos , Hipotiroidismo/sangre , Hipotiroidismo/inmunología , Tamizaje Masivo/métodos , Persona de Mediana Edad , Papel , Periodo Posparto , Valor Predictivo de las Pruebas , Embarazo , Complicaciones del Embarazo/sangre , Complicaciones del Embarazo/diagnóstico , Complicaciones del Embarazo/inmunología , Primer Trimestre del Embarazo , Diagnóstico Prenatal , Estudios Prospectivos , Reproducibilidad de los Resultados , Pruebas de Función de la Tiroides , Tiroiditis Autoinmune/sangre , Tiroiditis Autoinmune/inmunología , Tirotropina/inmunología , Adulto JovenRESUMEN
Phenylketonuria (PKU) is an autosomal recessive defect in hepatic metabolism of phenylalanine, which is secondary to mutations in the phenylalanine hydroxylase (PAH) gene. Sixty-seven ethnically Polish PKU patients, followed at the Outpatient Department of Pediatrics and Developmental Medicine in Poznan, Poland, were assessed for mutations in the PAH gene. Two mutations were identified in 61 of 67 patients and a single mutation was identified in the remaining six patients. The four most prevalent mutations (p.R408W, 68%; c.1066-11G>A, 6%; c.1315+1G>A, 5.2%; c.822-832delGCCCATGTATA, 3.7%) accounted for 83% of the mutant alleles. Fifteen additional mutations were identified of which most (13/15) were observed in an individual patient. Before knowledge of PAH genotypes, 19 patients were challenged with a 20 mg kg(-1) dose of 6R tetrahydrobiopterin (BH(4)) and serum phenylalanine concentration was monitored in hospital over 24 h. Two patients responded to the BH(4) challenge with a reduction of serum phenylalanine concentration >30% from baseline. PAH genotypes of the two responsive patients would have been predicted, as they contained mutations recognized as BH(4) responsive, whereas the 17 patients who were unresponsive would have been predicted as their mutations were either recognized as non-responsive or were highly deleterious frame-shift mutations. Overall, only 7.5% (5/ 67) of patients had PAH mutations recognized as responsive to co-factor therapy. Among the PKU patients from western Poland, PAH mutations responsive to BH(4) therapy are poorly represented; therefore, genotyping may be useful for identifying candidate patients likely to respond to BH(4) before physiological challenge.
Asunto(s)
Biopterinas/análogos & derivados , Mutación/genética , Fenilalanina Hidroxilasa/genética , Fenilcetonurias/genética , Adolescente , Adulto , Alelos , Biopterinas/uso terapéutico , Niño , Preescolar , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Lactante , Masculino , Linaje , Fenilalanina/sangre , Fenilcetonurias/tratamiento farmacológico , Polonia , Adulto JovenRESUMEN
The biochemical properties of mutant phenylalanine hydroxylase (PAH) enzymes and clinical characteristics of hyperphenylalaninaemic patients who bear these mutant enzymes were investigated. Biochemical characterization of mutant PAH enzymes p.D143G, p.R155H, p.L348V, p.R408W and p.P416Q included determination of specific activity, substrate activation, V(max), K(m) for (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin (BH(4)), K (d) for BH(4), and protein stabilization by BH(4). Clinical data from 22 patients either homozygous, functionally hemizygous, or compound heterozygous for the mutant enzymes of interest were correlated with biochemical parameters of the mutant enzymes. The p.L348V and p.P416Q enzymes retain significant catalytic activity yet were observed in classic and moderate PKU patients. Biochemical studies demonstrated that BH(4) rectified the stability defects in p.L348V and p.P416Q; additionally, patients with these variants responded to BH(4) therapy. The p.R155H mutant displayed low PAH activity and decreased apparent affinity for L-Phe yet was observed in mild hyperphenylalaninaemia. The p.R155H mutant does not display kinetic instability, as it is stabilized by BH(4) similarly to wild-type PAH; thus the residual activity is available under physiological conditions. The p.R408W enzyme is dysfunctional in nearly all biochemical parameters, as evidenced by disease severity in homozygous and hemizygous patients. Biochemical assessment of mutant PAH proteins, especially parameters involving interaction with BH(4) that impact protein folding, appear useful in clinical correlation. As additional patients and mutant proteins are assessed, the utility of this approach will become apparent.
Asunto(s)
Fenilalanina Hidroxilasa/genética , Fenilalanina Hidroxilasa/metabolismo , Fenilcetonurias/genética , Adulto , Biopterinas/análogos & derivados , Biopterinas/metabolismo , Dicroismo Circular , Escherichia coli/enzimología , Escherichia coli/genética , Fluorescencia , Humanos , Recién Nacido , Cinética , Mutagénesis Sitio-Dirigida , Proteínas Mutantes/análisis , Proteínas Mutantes/química , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Mutación Missense/fisiología , Organismos Modificados Genéticamente , Fenilalanina Hidroxilasa/análisis , Fenilalanina Hidroxilasa/química , Fenilcetonurias/enzimología , Fenilcetonurias/metabolismo , Fenilcetonurias/patología , Unión Proteica/genética , Pliegue de Proteína , Estabilidad Proteica , Adulto JovenRESUMEN
OBJECTIVE: Symptoms of hypothyroidism in adults can be mistaken for medical and psychiatric diseases, as well as for general signs of ageing such as weakness, lethargy and fatigue. The incidence of hypothyroidism is many-fold higher in adults than in newborn children. The latter have been routinely screened for the condition using filter paper dried blood spots (DBS) for nearly three decades but this cost-effective screening technique has only recently been applied to adults. This study was undertaken to show that DBS testing in adults and older children is an accurate way to screen for hypothyroidism. METHODS: Serum and DBS specimens were collected from adults and children. Assays were run on both specimens and the results correlated. In addition 972 specimens were collected from adults at community centres and nursing homes. Follow-up studies were performed on patients with positive results. RESULTS: The correlation coefficient for 118 matched serum and DBS specimens was 0.99. Thyroid-stimulating hormone (TSH) values were elevated in 50 of the 972 adults from nursing homes and community centres. Thirteen of these individuals were on thyroid medication and 28 had either high serum TSH or high thyroglobulin (TgAb) or thyroid peroxidase (TPOAb) antibody levels. CONCLUSIONS: Individuals can be screened for hypothyroidism by collecting finger stick DBS specimens at community centres, nursing homes and other locations which can be mailed by regular postal service to a central laboratory for accurate and inexpensive testing.
Asunto(s)
Hipotiroidismo/diagnóstico , Tirotropina/sangre , Adulto , Recolección de Muestras de Sangre/métodos , Preescolar , Humanos , Hipotiroidismo/prevención & control , Tamizaje Masivo/métodos , Persona de Mediana Edad , Papel , Análisis de Regresión , Reproducibilidad de los ResultadosAsunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Carnitina/análogos & derivados , Carnitina/sangre , Ácido Metilmalónico/sangre , Propionatos/sangre , Recolección de Muestras de Sangre , Reacciones Falso Positivas , Humanos , Recién Nacido , Papel , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
BACKGROUND: Deaths from inherited metabolic disorders may remain undiagnosed after postmortem examination and may be classified as sudden infant death syndrome. Tandem mass spectrometry (MS/MS) may reveal disorders of fatty acid oxidation in deaths of previously unknown cause. METHODS: We obtained filter-paper blood from 7058 infants from United States and Canadian Medical Examiners. Acylcarnitine and amino acid profiles were obtained by MS/MS. Specialized interpretation was used to evaluate profiles for disorders of fatty acid, organic acid, and amino acid metabolism. The analyses of postmortem blood specimens were compared with the analyses of bile specimens, newborn blood specimens, and specimens obtained from older infants at risk for metabolic disorders. RESULTS: Results on 66 specimens suggested diagnoses of metabolic disorders. The most frequently detected disorders were medium-chain and very-long-chain acyl-CoA dehydrogenase deficiencies (23 and 9 cases, respectively), glutaric acidemia type I and II deficiencies (3 and 8 cases, respectively), carnitine palmitoyl transferase type II/translocase deficiencies (6 cases), severe carnitine deficiency (4 cases), isovaleric acidemia/2-methylbutyryl-CoA dehydrogenase deficiencies (4 cases), and long-chain hydroxyacyl-CoA dehydrogenase/trifunctional protein deficiencies (4 cases). CONCLUSIONS: Postmortem metabolic screening can explain deaths in infants and children and provide estimates of the number of infant deaths attributable to inborn errors of metabolism. MS/MS is cost-effective for analysis of postmortem specimens and should be considered for routine use by Medical Examiners and pathologists in unexpected/unknown infant and child death.
Asunto(s)
Carnitina/análogos & derivados , Carnitina/sangre , Errores Innatos del Metabolismo/diagnóstico , Muerte Súbita del Lactante/diagnóstico , Autopsia , Bilis/química , Recolección de Muestras de Sangre , Canadá , Carnitina/análisis , Ácidos Grasos/metabolismo , Humanos , Lactante , Recién Nacido , Errores Innatos del Metabolismo/metabolismo , Oxidación-Reducción , Papel , Valores de Referencia , Espectrometría de Masa por Ionización de Electrospray , Estados UnidosRESUMEN
Medium-chain acyl-CoA dehydrogenase (MCAD) deficiency is the most frequently diagnosed mitochondrial beta-oxidation defect, and it is potentially fatal. Eighty percent of patients are homozygous for a common mutation, 985A-->G, and a further 18% have this mutation in only one disease allele. In addition, a large number of rare disease-causing mutations have been identified and characterized. There is no clear genotype-phenotype correlation. High 985A-->G carrier frequencies in populations of European descent and the usual avoidance of recurrent disease episodes by patients diagnosed with MCAD deficiency who comply with a simple dietary treatment suggest that MCAD deficiency is a candidate in prospective screening of newborns. Therefore, several such screening programs employing analysis of acylcarnitines in blood spots by tandem mass spectrometry (MS/MS) are currently used worldwide. No validation of this method by mutation analysis has yet been reported. We investigated for MCAD mutations in newborns from US populations who had been identified by prospective MS/MS-based screening of 930,078 blood spots. An MCAD-deficiency frequency of 1/15,001 was observed. Our mutation analysis shows that the MS/MS-based method is excellent for detection of MCAD deficiency but that the frequency of the 985A-->G mutant allele in newborns with a positive acylcarnitine profile is much lower than that observed in clinically affected patients. Our identification of a new mutation, 199T-->C, which has never been observed in patients with clinically manifested disease but was present in a large proportion of the acylcarnitine-positive samples, may explain this skewed ratio. Overexpression experiments showed that this is a mild folding mutation that exhibits decreased levels of enzyme activity only under stringent conditions. A carrier frequency of 1/500 in the general population makes the 199T-->C mutation one of the three most prevalent mutations in the enzymes of fatty-acid oxidation.
Asunto(s)
Acil-CoA Deshidrogenasas/deficiencia , Acil-CoA Deshidrogenasas/genética , Carnitina/análogos & derivados , Carnitina/sangre , Pruebas Genéticas/métodos , Mutación Missense/genética , Acil-CoA Deshidrogenasa , Acil-CoA Deshidrogenasas/química , Acil-CoA Deshidrogenasas/metabolismo , Alelos , Chaperonina 10/genética , Chaperonina 10/metabolismo , Chaperonina 60/genética , Chaperonina 60/metabolismo , Análisis Mutacional de ADN , Estabilidad de Enzimas , Escherichia coli/genética , Exones/genética , Haplotipos/genética , Heterocigoto , Homocigoto , Humanos , Recién Nacido , Espectrometría de Masas , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida/genética , Polimorfismo de Nucleótido Simple/genética , Pliegue de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Reproducibilidad de los Resultados , TemperaturaRESUMEN
CONTEXT: Collection of blood from newborns is a standard clinical procedure used for genetic screening. Typically, blood from a heel prick is absorbed onto standard collection paper and dried before analysis of metabolites, proteins, hormones, and more recently DNA. OBJECTIVE: To evaluate strategies to purify DNA for use with automated workstations. DESIGN: Two factors were used to evaluate several DNA purification protocols: residual heme contamination and amplification yield. The protocol that produced DNA with the lowest heme content and the highest amplification yield was selected. In combination with those two performance factors, the protocol with the fewest number of steps was chosen to reduce reagent use and processing time. SETTING: Industrial research and development laboratory. RESULTS: Robust amplification of DNA isolated from dried blood spots was demonstrated using both fluorescence and agarose gel-based detection methods. In addition, the samples had consistent DNA volumes and had no detectable cross-contamination. Suggested instrument settings, equipment, and supplies were included for automated processing of DNA from dried blood spots. CONCLUSION: A 4-step DNA processing protocol was developed for dried blood spots. The protocol could be performed in either a manual or automated format, making it possible to process hundreds of samples in 1 day.
Asunto(s)
ADN/aislamiento & purificación , Diagnóstico por Computador/normas , Pruebas Genéticas/métodos , Tamizaje Neonatal/métodos , Análisis Químico de la Sangre/métodos , ADN/sangre , ADN/química , Estudios de Evaluación como Asunto , Hemo/análisis , Humanos , Reacción en Cadena de la PolimerasaRESUMEN
Development of acylcarnitine and amino acid profiling using tandem mass spectrometry, and its application for use with dried blood specimens collected on filter-paper cards, has introduced an innovative new technology for detecting inborn errors of fatty acid, organic acid, and amino acid metabolism. From November 1, 1992 through June 30, 1999 we screened more than 700,000 newborns in Pennsylvania, Ohio, North Carolina, and Louisiana. We have prospectively detected 163 inborn errors of metabolism. Eighty-six patients have amino acid metabolism errors. Among them are phenylketonuria, hyperphenylalaninemia, maple syrup urine disease, and several urea cycle disorders. Thirty-two have organic acid metabolism errors, including glutaric aciduria type 1; 3-methylcrotonyl coenzyme A (CoA) carboxylase deficiency, propionic acidemia, methylmalonic acidemia, and 3-hydroxy-3-methylglutaryl-CoA lyase deficiency; and 45 have fatty acid oxidation errors, including 36 with medium-chain acyl-CoA dehydrogenase deficiency. Details of the methodology are presented and the potential of this screening technology is discussed.
Asunto(s)
Tamizaje Masivo/métodos , Espectrometría de Masas/métodos , Errores Innatos del Metabolismo/diagnóstico , Aminoácidos/análisis , Biomarcadores/análisis , Carnitina/análogos & derivados , Carnitina/análisis , Humanos , Recién Nacido , Espectrometría de Masas/normas , Errores Innatos del Metabolismo/prevención & control , Estudios ProspectivosRESUMEN
The aim of this study was to evaluate pancreatic function in total parenteral nutrition (TPN)-dependent children with permanent intestinal failure by measuring immunoreactive trypsinogen (IRT) levels. Between 1992 and 1996, 105 pediatric patients with permanent intestinal failure were referred to the Children's Hospital of Pittsburgh for small intestinal transplant evaluation. Serum samples were available from 55 of them. Ten suffered from intestinal pseudo-obstruction or microvillus inclusion disease, while 45 had short bowel syndrome (SBS). IRT levels were significantly higher (p < 0.001) in SBS patients (89.4 +/- 9.2 ng mL) compared to controls (43.4 +/- 5.6 ng/ nL) without liver, gastrointestinal, or kidney disease. IRT levels did not correlate with liver injury, length of bowel, or the cause of SBS. Five of 20 patients who underwent intestinal transplantation developed pancreatitis during a median post-operative follow up 15.4 months later. IRT levels failed to predict who would develop pancreatitis post-transplant. The data suggest that elevated plasma IRT levels are common among children with intestinal failure, but fail to identify patients at risk for pancreatitis post-transplant.
Asunto(s)
Enfermedades Intestinales/sangre , Intestinos/trasplante , Tripsinógeno/sangre , Adolescente , Niño , Preescolar , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Lactante , Enfermedades Intestinales/cirugía , Enfermedades Intestinales/terapia , Seudoobstrucción Intestinal/sangre , Seudoobstrucción Intestinal/cirugía , Seudoobstrucción Intestinal/terapia , Masculino , Pancreatitis/diagnóstico , Pancreatitis/etiología , Nutrición Parenteral Total , Factores de Riesgo , Síndrome del Intestino Corto/sangre , Síndrome del Intestino Corto/cirugía , Síndrome del Intestino Corto/terapia , Trasplante Homólogo/efectos adversosRESUMEN
Clinical and neonatal screening methods using a tandem mass spectrometer are clearly a model for modern laboratory testing in the new Millennium. By the year 2000, more than 1 million blood and plasma samples will have been tested in laboratories throughout the world for a battery of metabolic disorders using a tandem mass spectrometer as the primary analytical device. A tandem mass spectrometer is considered the "ultimate" analytical detector in a variety of biochemical and clinical methods because of its very high accuracy, selectivity, precision, versatility and robust nature. The ability to achieve very high and reproducible sample throughput (approximately 600 samples/instrument/24 h) has made this technology cost-effective for newborn screening. In order to reliably measure markers of inborn errors of metabolism while maintaining low costs and high efficiency, accuracy and quality, much attention needs to be placed on monitoring and maintenance of all components of the entire testing system. These components include specimen collection and sample preparation methods, analysis by LC tandem mass spectrometry, conversion of raw mass spectra (data) into clinically meaningful results (concentration), expert interpretation of these results so that the clinician can be provided with information to facilitate a diagnose, and follow-up and education so that the maximum benefits of newborn screening translate into prevention of disease symptoms or more effective treatments. Addressing each part of the whole system will produce a quality screening program that will detect a battery of disorders using tandem mass spectrometry with a disease frequency of nearly 1 in 4000 infants.
Asunto(s)
Espectrometría de Masas/tendencias , Errores Innatos del Metabolismo/diagnóstico , Tamizaje Neonatal/métodos , Técnicas de Laboratorio Clínico/instrumentación , Femenino , Predicción , Humanos , Recién Nacido , Masculino , Espectrometría de Masas/estadística & datos numéricos , Sensibilidad y Especificidad , Estados UnidosRESUMEN
Modern molecular biology, owing much to the Human Genome Initiative, has elucidated many of the genetic mechanisms underlying heritable metabolic disease. While the use of molecular methods has flourished in research laboratories, complexity and cost have limited their utility in newborn screening. Newborn blood cards provide high quality DNA samples able to provide reliable support to highly multiplexed polymerase chain reactions (PCR). New manufacturing processes have reduced the cost of DNA microarray technology to the point where it is a practical tool for population screening. In a single assay, a DNA microarray facilitates the co-detection of amplification products diagnostic for several genetic diseases. High throughput is achieved with automation at every step, from DNA extraction to detection of hybrids. We suggest that it is both feasible and practical to develop a first-tier newborn screening protocol based upon multiplex PCR and analysis of amplification products using DNA microarrays. Initial data utilizing the model systems of sickle cell disease, alpha-1-antitrypsin deficiency and Factor V Leiden will be reported.
Asunto(s)
ADN/análisis , Tamizaje Neonatal/métodos , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Reacción en Cadena de la Polimerasa/métodos , ADN/genética , Femenino , Humanos , Recién Nacido , Masculino , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The incidence of inborn errors of metabolism (IEM) in Thailand is yet unknown. However, by estimation it is generally accepted to be 1 in 5,000. From a survey in 7 medical schools from different parts of the country and a large pediatric hospital in Bangkok, we found numerous cases of IEM nationwidc. Thesc were amino acid disorders, carbohydrate disorders, urea cycle defects, peroxisomal, lysosomal storage disorders, and many others. Since Thais are quite homogeneous in their genetic make-up; it is, therefore, very likely that IEM is much more prevalent than we realized. With the exception of thalassemias, IFAM is probably very common in Thailand and other countries in the Asia-Pacific region. IEM identified were amino acid disorders eg phenylketonuria, maple syrup urine disease: urea cycle disorders eg ornithine transcarbamylase deficincy (OTC), argininosuccinic lyase deficiency (ALD), argininosuccinic acid synthetasc deficiency (ASD); glycogen storage disorders eg Pompe's discase, Von Gierkc's; organic acid disorders eg, isovaleric acidemia, methylmalonic acidemia. Lysosmal storagc disorders identified were GM1 gangliosidosis, mucolipidosis II, Hurler, Hunter, Maroteaux-Lamy, Sialidosis (neuraminidase deficicncy), Sly, Scheie, Gaucher, Niemann-Pick, Sandhoff and many other neurodegeneraative disorders identified were rhizomelic chondrodysplasia punctata (RCDP) and Zellweger. Recently fatty acid oxidation disorders: MCAD, translocase deficiency and multiple carbosxylase deficiency 9biotinidase deficiency) were also identified.
Asunto(s)
Errores Innatos del Metabolismo/diagnóstico , Niño , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Errores Innatos del Metabolismo/epidemiología , Tailandia/epidemiologíaRESUMEN
From a retrospective study in Medical Genetics Unit, Department of Pediatrics, Siriraj Hospital Faculty of Medicine, Mahidol University in Bangkok (1983-1988), the estimated pediatric patients with clinically suspected IEM are approximately 2-4% of total annual pediatrics admission of 5,000 or more. This is, a low estimation since survey from all teaching hospitals in the country including the largest Children's Hospital in Bangkok indicated the presence of numerous IEM. However, most IEM were clinically diagnosed with limited laboratory facilities. We started a collaboration with Magee Womens Hospital of Pittsburgh and NeoGen Screening, USA; using tandem mass spectrometry to diagnose high risk infants and children for IEM from July 1993 to March 1998. Of total 146 samples sent, we detected numerous metabolic disorders (11.2%) eg phenylketonuria, organic acidemia, maple syrup urine disease, isovaleric acidemia, methylmalonic acidemia, albinism, translocase/carnitine palmitoyltransferase type II, G6PD deficiency and lysinuric protein intolerance.
Asunto(s)
Análisis Químico de la Sangre , Errores Innatos del Metabolismo/diagnóstico , Espectrometría de Masa por Ionización de Electrospray , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Aminoácidos/sangre , Análisis Químico de la Sangre/métodos , Carnitina Aciltransferasas/deficiencia , Niño , Ácidos Grasos/metabolismo , Hemiterpenos , Humanos , Lactante , Recién Nacido , Masculino , Enfermedad de la Orina de Jarabe de Arce/diagnóstico , Ácidos Pentanoicos/sangre , Fenilcetonurias/diagnóstico , Estudios Retrospectivos , TailandiaRESUMEN
Isolated 3-methylcrotonyl coenzyme A carboxylase (MCC) deficiency was documented in four adult women from the Amish/Mennonite population of Lancaster County, Pennsylvania. Metabolic and enzymatic investigations in these individuals were instituted after the detection of abnormal acylcarnitine profiles in blood spots obtained from their newborn children, in whom MCC activity was normal.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Ligasas de Carbono-Carbono/deficiencia , Leucina/metabolismo , Adulto , Ligasas de Carbono-Carbono/sangre , Carnitina/sangre , Preescolar , Cristianismo , Femenino , Humanos , Recién Nacido , Madres , Pennsylvania , Espectrometría de Masa Bombardeada por Átomos VelocesRESUMEN
We report the application of tandem mass spectrometry to prospective newborn screening for medium-chain acyl-CoA dehydrogenase (MCAD) deficiency. MCAD deficiency is diagnosed from dried blood spots on filter paper cards from newborns on the basis of the increase of medium chain length acylcarnitines identified by isotope dilution mass spectrometry methods. A robust and accurate semiautomated method for the analysis of medium chain length acylcarnitines as their butyl esters was developed and validated. Quantitative data from the analyses of 113 randomly collected filter paper blood spots from healthy newborns showed low concentrations of medium chain length acylcarnitines such as octanoylcarnitine. The maximum concentration of octanoylcarnitine was 0.22 mumol/L, with the majority being at or below the detection limit. In all 16 blood spots from newborns with confirmed MCAD deficiency, octanoylcarnitine was highly increased [median 8.4 mumol/L (range 3.1-28.3 mumol/L)], allowing easy detection. The concentration of octanoylcarnitine was significantly higher in these 16 newborns (< 3 days of age) than in 16 older patients (ages 8 days to 7 years) with MCAD deficiency (median 1.57 mumol/L, range 0.33-4.4). The combined experience of prospective newborn screening in Pennsylvania and North Carolina has shown a disease frequency for MCAD deficiency of 1 in 17,706. No false-positive and no known false-negative results have been found. A validated method now exists for prospective newborn screening for MCAD deficiency.
Asunto(s)
Acil-CoA Deshidrogenasas/deficiencia , Carnitina/análogos & derivados , Acil-CoA Deshidrogenasa , Butanoles , Carnitina/sangre , Humanos , Recién Nacido , Espectrometría de Masas , Estudios Prospectivos , Técnica de Dilución de Radioisótopos , Reproducibilidad de los Resultados , Factores de TiempoRESUMEN
In this study, time-of-flight secondary ion mass spectrometry was used to distinguish between blood of normal infants and that of individuals with Smith-Lemli-Opitz (SLO) syndrome. SLO syndrome results in an abnormally low concentration of blood cholesterol and an elevated concentration of 7-dehydrocholesterol. Blood was spotted on filter paper and analyzed directly with no extractions or separations. Results showed that using ratios of fragment ions for cholesterol/dehydrocholesterol, patients with SLO and normal individuals could be unambiguously distinguished. Unknown samples from 28 individuals were obtained and identified correctly.
Asunto(s)
Síndrome de Smith-Lemli-Opitz/diagnóstico , Espectrometría de Masa de Ion Secundario , Adulto , Niño , Preescolar , Colesterol/sangre , Deshidrocolesteroles/sangre , Estudios de Evaluación como Asunto , Filtración , Humanos , Lactante , Recién Nacido , Tamizaje Masivo , Síndrome de Smith-Lemli-Opitz/sangreRESUMEN
Postoperative patients are hypercatabolic. They also suffer a degree of gastrointestinal (GI) dysfunction impairing nutritional intake. Safe enteral absorption had been limited to a maximum of 500 kcal provided over the initial 24 h in all previously reported regimens. Several days of negative nitrogen balance and diminution of serum branched-chain amino acids (BCAA) result. Some data suggest an association with suboptimal wound healing, and immune competence. We tested the hypothesis that immediate exploitation of more effectively preserved GI function could prevent this depressed serum concentration within hours of surgery. Our study group consisted of 34 consecutive elective "open" cholecystectomy patients who had terminal esophageal, gastric, and proximal duodenal decompression. Simultaneous distal duodenal feeding of elemental diet began immediately in the Recovery Room at 300 mL/h for 8-16 h, providing 2,400-4,800 kcal and 100-200 g amino acids. We found that each serum BCAA concentration rose above basal by one hour. The differences reached statistical significance (p < or = 0.05) within 2 h after surgery, when leucine had risen above basal levels by 70%, isoleucine by 63%, and valine by 26%. Elevations in these BCAA serum concentrations persisted for the duration of feeding. GI function can be maintained and successfully utilized in the immediate postoperative period for absorption of 300 kcal/h. Postoperative depression of serum BCAA concentrations are prevented. The serum BCAA rise of these GI protected and immediately fed postoperative patients contrasts with the 48-72 h decline universally reported with conventional protein- and calorie-deficient regimens. The optimum nutritional regimen for achievement of clinically significant enhanced wound healing, sepsis resistance, and muscle sparing, remains to be determined.
Asunto(s)
Aminoácidos de Cadena Ramificada/sangre , Descompresión Quirúrgica , Procedimientos Quirúrgicos del Sistema Digestivo , Nutrición Enteral , Adulto , Anciano , Anciano de 80 o más Años , Colecistectomía , Femenino , Humanos , Isoleucina/sangre , Cinética , Leucina/sangre , Masculino , Persona de Mediana Edad , Periodo Posoperatorio , Valina/sangreRESUMEN
Medium chain acyl-CoA dehydrogenase deficiency (MCAD) is a defect in the mitochondrial oxidation of fatty acids. The disorder typically presents with episodes of vomiting and hypoglycemia, sometimes with changes in mental status and hepatic failure. These Reye's-like features may culminate in coma and death. Stress, intercurrent illness, and reaction to childhood immunization have been shown to precipitate acute metabolic episodes in MCAD patients. All cases are caused by mutations of the single MCAD gene on chromosome 1. Most clinically ascertained cases are caused by an A985G transition in exon 11. Here we report the preliminary findings of MCAD patients detected prospectively through a supplemental newborn screening program in Pennsylvania using tandem mass spectrometry. From the first 80,371 newborns screened we prospectively found nine babies with MCAD (1/8930) plus two additional newborns screened because of a previously known family history. Molecular analysis showed 56% of the detected patients to be compound heterozygotes for the A985G and a second mutation. This is in contrast to clinical retrospective studies which have found only 20% to be compound heterozygotes. We have identified two of the other mutations including a novel mutation (DG91/C92, 6-bp deletion) in one of our patients by using single-stranded conformation polymorphism (SSCP) and sequence analysis of conformers. Our results confirm that MCAD is one of the more common inborn errors of metabolism. The different mutation frequencies observed between retrospective clinical studies and our prospective newborn screening study suggest that clinical ascertainment may lead to preferential identification of the A985G mutation.
Asunto(s)
Acil-CoA Deshidrogenasas/deficiencia , Acil-CoA Deshidrogenasa , Acil-CoA Deshidrogenasas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Carnitina/análogos & derivados , Carnitina/sangre , Estudios de Cohortes , ADN/análisis , Femenino , Eliminación de Gen , Pruebas Genéticas , Heterocigoto , Humanos , Incidencia , Recién Nacido , Masculino , Datos de Secuencia Molecular , Mutación , Pennsylvania , Estudios ProspectivosRESUMEN
We report a new method for the diagnosis of maple syrup urine disease (MSUD) from dried blood spots on newborn screening cards based on tandem mass spectrometry (MS-MS). The mean +/- SD concentration of Leu plus Ile in normal newborns was 151 +/- 47 mumol/L (n = 1096); for Val, 131 +/- 58 mumol/L (n = 791). SDs were lower when the concentrations of these amino acids were expressed relative to that of Phe. The mean ratio for Leu + Ile to Phe was 2.5 +/- 0.49; for Val to Phe, 2.18 +/- 0.51. These results compare well with values previously reported in the literature. With these criteria, samples from a collection categorized by a bacterial inhibition assay as normal or falsely positive for MSUD were normal by MS-MS [(Leu + Ile): Phe < 5.0]. Samples from confirmed MSUD patients were categorized as abnormal [(Leu+Ile): Phe > 9.0] by MS-MS.