RESUMEN

We analysed samples of 400 Finnish males using nine Y-chromosomal short tandem repeat (STR) loci (minimal haplotype); for 200 of these subjects an additional seven Y-chromosomal STR loci were used. The geographical distribution of the observed haplotypes was determined from 200 individuals of known paternal origin within Finland. The observed number of alleles varied from 2 to 13 alleles per locus. A total of 146 minimal haplotypes were identified in our population sample. Interestingly, 90 (22.5%) individuals shared an identical haplotype. This haplotype was extremely frequent in the northern and eastern subpopulations of Savo, Pohjanmaa and Karjala (53, 42 and 37%, respectively). With the seven additional loci analysed in the sample of 200 individuals, 120 haplotypes were identified, and individuals sharing the most common haplotype decreased to 13.0%. However, in comparison to other European populations, the Finnish population showed decreased genetic diversity (GD) when the number of different minimal haplotypes in the population was divided by the sample size (36.5% in Finns versus 83.7% on average). Our results strongly support the earlier hypothesis of individual isolated Y-chromosomal lineages and population substructuring in Finland. For paternity testing, power of exclusion was 92% using minimal haplotype data, but including the seven additional loci this value increased to 97%.

Asunto(s)

Cromosomas Humanos Y , Variación Genética , Genética de Población , Haplotipos , Secuencias Repetidas en Tándem , Adulto , Dermatoglifia del ADN/métodos , Finlandia , Frecuencia de los Genes , Humanos , Masculino , Persona de Mediana Edad , Paternidad

RESUMEN

OBJECTIVE: To evaluate the clinical significance of antifilaggrin antibodies (AFA) measured by an enzyme linked immunosorbent assay (ELISA) in serial specimens from patients with recent onset rheumatoid arthritis (RA). METHODS: Filaggrin was purified from human skin and used as an antigen in ELISA. The AFA test was applied to five serial specimens from 78 patients with recent onset RA followed up for three years. Rheumatoid factor (RF) had been measured earlier from the same samples by quantitative immunoturbidimetry. RESULTS: The mean AFA level was highest at entry (54% positive), followed by a statistically significant decline at six months and a slight increase at three years. AFA were persistently positive in 23 patients and persistently negative in 28 patients. Eleven of the latter patients were persistently negative for RF. At study entry AFA levels correlated to some degree with RF levels. In general, raised AFA levels at entry were associated with an active and treatment resistant disease, but they did not predict radiological progression. CONCLUSIONS: The test for AFA has potential for an additional immunological test for RA.

Asunto(s)

Artritis Reumatoide/inmunología , Autoanticuerpos/sangre , Proteínas de Filamentos Intermediarios/inmunología , Adulto , Anciano , Biomarcadores/sangre , Ensayo de Inmunoadsorción Enzimática , Epidermis/inmunología , Femenino , Proteínas Filagrina , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Factor Reumatoide/sangre , Índice de Severidad de la Enfermedad

RESUMEN

We evaluated the sensitivity and prognostic value of an enzyme-linked immunosorbent assay (ELISA) for the measurement of antifilaggrin antibodies (AFA), using filaggrin purified from human skin as an antigen. The AFA test was applied to a series of 306 patients with various recent-onset inflammatory joint diseases. The results were compared to those of the conventional immunofluorescence tests for antikeratin antibody (AKA) and antiperinuclear factor (APF) and of the rheumatoid factor (RF) tests from a previous study. There was a very good agreement between the results of the tests for APF and AFA (kappa-value 0.79 in patients with peripheral poly/oligoarthritis). The agreement between the tests for AKA and AFA was significant but less pronounced (kappa-value 0.50). The AFA test detected 10/22 of the RF-negative erosive cases, particularly those with a large number of erosive joints. Thus, the test for AFA supplements RF in the prediction of erosiveness.

Asunto(s)

Artritis Reumatoide/diagnóstico , Artritis Reumatoide/inmunología , Epidermis/química , Proteínas de Filamentos Intermediarios/inmunología , Adulto , Anciano , Anticuerpos Antinucleares/sangre , Ensayo de Inmunoadsorción Enzimática , Proteínas Filagrina , Estudios de Seguimiento , Humanos , Inmunoglobulina G/sangre , Proteínas de Filamentos Intermediarios/análisis , Queratinas/inmunología , Modelos Logísticos , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Pronóstico , Factor Reumatoide/sangre , Sensibilidad y Especificidad

RESUMEN

We have analysed close to 30,000 human germline transmission events at five microsatellite loci (D3S1359, HumTH01, HumvWA, HumTPO and HumFES) and four minisatellite loci (D1S80, ApoB, Col2A1 and D17S30). At these loci the mutation rates are similar at the microsatellite and the minisatellite loci, varying from 0.2 x 10(-3) to < 3.3 x 10(-3) and from 0.5 x 10(-3) to 1.5 x 10(-3), respectively. Interestingly, paternal mutations appeared to be dominant at the microsatellite loci, whilst maternal mutations are dominant at minisatellite loci. Based on our data, no unequivocal support for a strict strand-slippage mutation mechanism (gain or loss of a single repeat) was found, although the vast majority of the mutational events were small gains or losses of one to three repeats, and only few unequivocal large gains or losses were observed.

Asunto(s)

Mutación de Línea Germinal , Repeticiones de Microsatélite , Alelos , Humanos

RESUMEN

Mutations in the FBN1 gene cause Marfan syndrome (MFS), a dominantly inherited connective tissue disease. Almost all the identified FBN1mutations have been family specific, and the rate of new mutations is high. We report here a de novo FBN1mutation that was identified in two sisters with MFS born to clinically unaffected parents. The paternity and maternity were unequivocally confirmed by genotyping. Although one of the parents had to be an obligatory carrier for the mutation, we could not detect the mutation in the leukocyte DNA of either parent. To identify which parent was a mosaic for the mutation we analyzed several tissues from both parents, with a quantitative and sensitive solid-phase minisequencing method. The mutation was not, however, detectable in any of the analyzed tissues. Although the mutation could not be identified in a sperm sample from the father or in samples of multiple tissue from the mother, we concluded that the mother was the likely mosaic parent and that the mutation must have occurred during the early development of her germ-line cells. Mosaicism confined to germ-line cells has rarely been reported, and this report of mosaicism for the FBN1 mutation in MFS represents an important case, in light of the evaluation of the recurrence risk in genetic counseling of families with MFS.

Asunto(s)

Mutación de Línea Germinal/genética , Síndrome de Marfan/genética , Proteínas de Microfilamentos/genética , Mosaicismo/genética , Adulto , Alelos , Niño , Preescolar , Análisis Mutacional de ADN , Exones/genética , Padre , Femenino , Fibrilina-1 , Fibrilinas , Fibroblastos/metabolismo , Fibroblastos/patología , Haplotipos/genética , Humanos , Leucocitos/metabolismo , Leucocitos/patología , Masculino , Síndrome de Marfan/diagnóstico , Síndrome de Marfan/patología , Madres , Mutación Missense/genética , Linaje , Polimorfismo Conformacional Retorcido-Simple , Embarazo , Diagnóstico Prenatal

RESUMEN

BACKGROUND: The so-called antikeratin antibody (AKA) and the antiperinuclear factor (APF) that recognize proteins related to human epidermal filaggrin belong to the most specific serological markers of rheumatoid arthritis (RA). However, assays for the detection of AKA and APF are currently based on immunofluorescence, a method that is subject to arbitrary interpretation and inadequate standardization of the substrates. METHODS: Proteins extracted from human epidermis were separated by reversed-phase high-performance liquid chromatography (HPLC). Filaggrin-containing fractions, identified in immunoblotting by monoclonal antifilaggrin antibodies, were then subjected to gel filtration HPLC and, finally, to a second reversed-phase HPLC step. Tryptic digestion, amino acid sequencing and mass spectrometry were used to confirm the identity of the purified protein. Filaggrin was used as antigen in enzyme-linked immunosorbent assay (ELISA) to measure IgG class antifilaggrin antibodies. RESULTS: The filaggrin preparation obtained gave a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, binding monoclonal antifilaggrin antibody in immunoblotting. Amino acid sequences of all 10 tryptic peptides analyzed were shown to originate from human filaggrin. Antifilaggrin antibody levels exceeded the 99th percentile level of 100 middle-aged blood donors in 26/55 (47%) RA sera. At a similar cutoff level 28/55 (51%) of the RA sera were positive in the AKA test. Of the 26 antifilaggrin-positive sera, 21 were also AKA-positive. CONCLUSION: Human filaggrin can be purified by standard biochemical techniques, despite the heterogeneity of the protein, and used in ELISA for testing autoantibodies to filaggrin. The sensitivity of the assay equals that of the AKA test.

Asunto(s)

Artritis Reumatoide/inmunología , Autoanticuerpos/sangre , Epidermis/química , Inmunoglobulina G/sangre , Proteínas de Filamentos Intermediarios/inmunología , Proteínas de Filamentos Intermediarios/aislamiento & purificación , Adulto , Anciano , Secuencia de Aminoácidos , Artritis Reumatoide/sangre , Cromatografía Líquida de Alta Presión , Ensayo de Inmunoadsorción Enzimática , Femenino , Proteínas Filagrina , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido

RESUMEN

We have developed a triplex PCR method for D3S1359, HumTH01 and HumTPO tetranucleotide loci and a duplex PCR method for HumFES/FPS and HumvWA31A tetranucleotide loci using high resolution polyacrylamide gel electrophoresis and silver staining. The methods were evaluated for paternity testing and individual identification and allele frequencies at these loci are reported for 189-3387 unrelated individuals in the Finnish population. The D3S1359 locus, especially, was found to be a highly informative locus. Seventeen alleles were found in the D3S1359 locus with a highest observed allele frequency of 0.199, a high exclusion power (PE) in paternity testing (0.78) and a high observed heterozygosity (0.89). The combined PE for these five loci was 0.99.

Asunto(s)

Paternidad , Reacción en Cadena de la Polimerasa/métodos , Secuencias Repetitivas de Ácidos Nucleicos , Alelos , Finlandia , Genotipo , Humanos , Masculino

RESUMEN

The genetic relationships between two Finno-Ugric-speaking populations, the Finns and the Finnish Saami (Lapps), were studied by using PCR for six nuclear-DNA marker loci, mitochondrial restriction-site polymorphism, and sequence variation of a 360-bp segment of the mitochondrial control region. The allele frequencies of each of the nuclear-DNA marker loci and the frequencies of mtDNA restriction haplotypes were significantly different between the populations. The Saami showed exceptionally low variation in their mtDNA restriction sites. The 9-bp deletion common in East Asian populations was not observed, nor did the haplotype data fit into the haplogroup categorization of Torroni et al. The average number of nucleotide substitutions from the mtDNA haplotype data indicated that the Finnish Saami may be closer to the Finns than to the other reference populations, whereas nuclear DNA suggested that the Finns are more closely related to the European reference populations than to the Finnish Saami. The similarity of the Finns to the other Europeans was even more pronounced according to the sequence data. We were unable to distinguish between the Finns and either the Swiss or Sardinian reference populations, whereas the Finnish Saami clearly stood apart. The Finnish Saami are distinct from other Circumarctic populations, although two of the lineages found among the Saami showed closer relationship to the Circumarctic than to the European lineages. The sequence data indicated an exceptionally high divergence for the Saami mtDNA control lineages. The distribution of the pairwise nucleotide differences in the Saami suggested that this population has not experienced an expansion similar to what was indicated for the Finns and the reference populations.

Asunto(s)

ADN Mitocondrial/genética , ADN/genética , Etnicidad/genética , Filogenia , Secuencia de Bases , Núcleo Celular , Finlandia , Marcadores Genéticos , Variación Genética , Haplotipos , Humanos , Lenguaje , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Grupos Raciales/genética , Mapeo Restrictivo , Eliminación de Secuencia

RESUMEN

Antiphospholipid antibodies in autoimmune sera have been shown to react with a complex of phospholipids (cardiolipin) and a plasma phospholipid-binding protein, beta 2-glycoprotein I (apolipoprotein H). The binding of these antibodies was inhibited by oxidized low-density lipoprotein (LDL) in sera from patients with systemic lupus erythematosus (SLE), suggesting cross-reactivity between antiphospholipid antibodies and antibodies binding to oxidized LDL. We purified antiphospholipid antibodies by cardiolipin-polyacrylamide column from seven SLE sera and studied the reactivity of eluted fractions with cardiolipin-beta 2-glycoprotein I complex and oxidized LDL (malondialdehyde-conjugated LDL) in solid-phase enzyme immunoassay. In four sera the binding of IgG antibodies to cardiolipin-beta 2-glycoprotein I complex and to oxidized LDL appeared in the same fractions, whereas in three sera reactivities against cardiolipin and oxidized LDL were observed, at least in part, in separate fractions. The binding to solid-phase cardiolipin was dependent on the presence of exogenous beta 2-glycoprotein I in all fractions. Our findings show that antiphospholipid antibodies are heterogeneous in their binding to oxidized LDL, indicating that these two antibodies may have different subspecificities. Some eluted fractions reacted only with oxidized LDL, and did not show binding to cardiolipin-beta 2-glycoprotein I complex, suggesting that the lipid part in the antigenic complex might be responsible for the cross-reactivity of these antibodies. Accordingly, the biological functions of antibodies against phospholipid-beta 2-glycoprotein I complex and antibodies against oxidized LDL may also be different.

Asunto(s)

Anticuerpos Anticardiolipina/química , Anticuerpos Anticardiolipina/aislamiento & purificación , Afinidad de Anticuerpos , Lipoproteínas LDL/química , Lipoproteínas LDL/inmunología , Adulto , Anciano , Apolipoproteínas/inmunología , Reacciones Cruzadas , Glicoproteínas/inmunología , Humanos , Lipoproteínas LDL/metabolismo , Persona de Mediana Edad , Oxidación-Reducción , beta 2 Glicoproteína I

RESUMEN

When mitochondrial DNA sequence variation is analyzed from a sample of 637 individuals in 14 European populations, most populations show little differentiation with respect to each other. However, the Saami distinguish themselves by a comparatively large amount of sequence difference when compared with the other populations, by a different distribution of sequence diversity within the population, and by the occurrence of particular sequence motifs. Thus, the Saami seem to have a long history distinct from other European populations. Linguistic affiliations are not reflected in the patterns of relationships of mitochondrial lineages in European populations, whereas prior studies of nuclear gene frequencies have shown a correlation between genetic and linguistic evolution. It is argued that this apparent contradiction is attributable to the fact that genetic lineages and gene frequencies reflect different time perspectives on population history, the latter being more in concordance with linguistic evolution.

Asunto(s)

ADN Mitocondrial/genética , Etnicidad/genética , Genoma Humano , Lenguaje , Secuencia de Bases , Etnicidad/clasificación , Etnicidad/historia , Europa (Continente) , Evolución Molecular , Finlandia , Frecuencia de los Genes , Marcadores Genéticos , Historia Moderna 1601- , Humanos , Datos de Secuencia Molecular , Homología de Secuencia de Ácido Nucleico

RESUMEN

BACKGROUND: There is general agreement that proteins eluting from different natural rubber latex products can cause immediate type hypersensitivity reactions in latex-allergic patients. However, there is as yet no consensus as to what are the most important allergens in natural rubber latex. OBJECTIVE: We wanted to purify and characterize at the primary structure level three natural latex proteins, suggested to represent significant allergens. METHODS: Proteins were purified from ultracentrifuged bottom fraction of natural rubber latex using high performance liquid chromatography gel filtration and reversed phase chromatography. Purified proteins were subjected to tryptic cleavage, peptide separation and amino acid sequencing. Immunoblotting was used to demonstrate IgE antibodies to the purified proteins in sera from latex-allergic patients. RESULTS: A 20 kDa protein was identified by amino acid sequencing as prohevein, a major protein in the rubber tree Hevea brasiliensis, and a 30 kDa natural rubber latex protein as hevamine, another essential rubber tree protein. A third, previously undescribed natural rubber latex protein, showed high homology to several plant endo-1,3-beta-glucosidases. In immunoblotting, the purified prohevein bound IgE antibodies from 24/29 (83%) sera of latex-allergic patients including positive results in 4/6 latex-allergic children with spina bifida or other congenital anomalies. The purified prohevein elicited positive skin-prick test reactions in all six latex-allergic patients showing IgE to prohevein. The purified 36 kDa protein bound IgE from 6/29 (21%) latex-allergic sera, and the purified hevamine from only 1/29 patient sera. CONCLUSION: The observed high frequency of IgE antibodies to prohevein suggests that this protein is a major natural rubber latex allergen.

Asunto(s)

Alérgenos/aislamiento & purificación , Péptidos Catiónicos Antimicrobianos , Látex/inmunología , Lectinas/inmunología , Lectinas de Plantas , Proteínas de Plantas/inmunología , Precursores de Proteínas/inmunología , Adolescente , Adulto , Secuencia de Aminoácidos , Niño , Femenino , Humanos , Immunoblotting , Lectinas/aislamiento & purificación , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Proteínas de Plantas/aislamiento & purificación , Precursores de Proteínas/aislamiento & purificación

RESUMEN

We purified from natural rubber latex (NRL) by means of high-performance liquid chromatography a 27-kD protein, recognized characteristically by IgE in sera from latex-allergic children with spina bifida or other congenital anomalies and histories of multiple surgeries. N-terminal sequence analysis of the purified 27-kD protein was unsuccessful suggesting that its N-terminus is blocked. To obtain internal sequence information from the protein it was digested with trypsin and the purified tryptic peptides were subjected to sequence analysis. Thirteen of the 14 sequenced peptides revealed no significant homology to any of the published protein sequences indicating that the 27-kD protein is previously undescribed at the primary structure level. However, one of the 14 sequenced peptides showed significant homology to the rubber elongation factor, a 14.6-kD NRL protein. For the time being, the 27-kD NRL protein is the first molecularly characterized NRL allergen associated with defined clinical manifestations of latex allergy.

Asunto(s)

Alérgenos/química , Alérgenos/aislamiento & purificación , Látex/química , Látex/aislamiento & purificación , Disrafia Espinal/inmunología , Adolescente , Adulto , Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Electroforesis en Gel Bidimensional , Electroforesis en Gel de Poliacrilamida , Femenino , Humanos , Immunoblotting , Inmunoglobulina E/inmunología , Masculino , Datos de Secuencia Molecular

RESUMEN

The allele frequencies at the tetranucleotide repeat (TCTA) vWA locus in the vWF gene were determined in the general Finnish population, in a population representing an internal isolate of Finland, in the Vologda-Russian population, and in US Black population samples. The allele and genotype frequencies from these population samples were compared with each other and with those reported from Spanish and British population samples. Statistically significant differences were demonstrated between most of the different groups (Finns vs. Vologda-Russians, Finns vs. US Blacks, Finns vs. Spanish, Vologda-Russians vs. US Blacks, Vologda-Russians vs. Spanish, US Blacks vs. Spanish and US Blacks vs. British Caucasians), but not between the two Caucasoid population samples from Finland and Great Britain, nor between or within the subpopulation samples from Finland and those from Vologda-Russia. In addition, the vWA marker was evaluated and demonstrated to be reliable for forensic purposes and paternity testing.

Asunto(s)

Población Negra/genética , ADN Satélite/análisis , Medicina Legal/métodos , Frecuencia de los Genes/genética , Población Blanca/genética , Factor de von Willebrand/genética , Adulto , Alelos , Animales , Secuencia de Bases , Niño , Femenino , Finlandia , Marcadores Genéticos , Humanos , Masculino , Datos de Secuencia Molecular , Pan troglodytes , Paternidad , Polimorfismo Genético , Secuencias Repetitivas de Ácidos Nucleicos , Federación de Rusia , Estados Unidos

RESUMEN

A 28-year-old man with chronic myeloid leukaemia received an allogeneic bone marrow transplant after conditioning with daunorubicin, cyclophosphamide and fractionated total body irradiation (TBI). Four years later his wife gave birth to a healthy child. Although the patient was azospermic serologic HLA testing suggested that the patient was the father of the child. DNA fingerprinting as well as analysis of three variable number of tandem repeats (VNTR) loci D1S80 (MCT118), D17S30 (YNZ22) and the apolipoprotein B hypervariable region (apo B 3') gave unequivocal results showing that the patient was the father. Fathering a child after TBI-containing regimen has been very rare and this is the first case where the paternity has been proven with DNA methodology.

Asunto(s)

Trasplante de Médula Ósea , Dermatoglifia del ADN , ADN/análisis , Oligospermia/etiología , Paternidad , Traumatismos por Radiación/etiología , Irradiación Corporal Total , Adulto , Ciclofosfamida/efectos adversos , Daunorrubicina/efectos adversos , Femenino , Humanos , Hipogonadismo/etiología , Recién Nacido , Leucemia Mieloide de Fase Crónica/terapia , Masculino , Embarazo , Inducción de Remisión , Espermatogénesis/efectos de la radiación , Irradiación Corporal Total/efectos adversos

RESUMEN

We have developed a new method for forensic identification of individuals, in which a panel of biallelic DNA markers are amplified by the PCR, and the variable nucleotides are detected in the amplified DNA fragments by the solid-phase minisequencing method. A panel of 12 common polymorphic nucleotides located on different chromosomes with reported allele frequencies close to .5 were chosen for the test. The allele frequencies for most of the markers were found to be similar in the Finnish and other Caucasian populations. We also introduce a novel approach for rapid determination of the population frequencies of biallelic markers. By this approach we were able to determine the allele frequencies of the markers in the Finnish population, by quantitative analysis of three pooled DNA samples representing 3,000 individuals. The power of discrimination and exclusion of the solid-phase minisequencing typing test with 12 markers was similar to that of three VNTR markers that are routinely used in forensic analyses at our institute. The solid-phase minisequencing method was successfully applied to type paternity and forensic case samples. We also show that the quantitative nature of our method allows typing of mixed samples.

Asunto(s)

Alelos , ADN/genética , Marcadores Genéticos , Reacción en Cadena de la Polimerasa , Análisis de Secuencia/métodos , Adulto , Secuencia de Bases , Niño , Femenino , Medicina Legal/métodos , Frecuencia de los Genes , Genotipo , Humanos , Masculino , Datos de Secuencia Molecular , Paternidad , Secuencias Repetitivas de Ácidos Nucleicos , Estadística como Asunto

RESUMEN

We describe a method for DNA-typing, in which a panel of biallelic markers are detected by the solid-phase minisequencing method (Syvänen et al., 1990). This method identifies single nucleotide variations in DNA fragments amplified by the PCR. Determination of the panel of 12 markers selected in this study proved to be an efficient and reliable method for forensic identification of individuals. We also introduce a novel approach for rapid determination of allele frequencies by quantitative analysis of pooled DNA samples.

Asunto(s)

Dermatoglifia del ADN/métodos , Medicina Legal , Alelos , Dermatoglifia del ADN/estadística & datos numéricos , ADN Satélite/genética , Estudios de Evaluación como Asunto , Frecuencia de los Genes , Marcadores Genéticos , Humanos , Masculino , Paternidad , Secuencias Repetitivas de Ácidos Nucleicos , Análisis de Secuencia de ADN/métodos

RESUMEN

The effect of a stacking gel, the pH and crosslinking agent concentration on the resolution and sharpness of PCR amplified VNTR alleles in a vertical discontinuous polyacrylamide gel electrophoresis system was investigated. The experiments show that the use of a low crosslinking agent concentration, a stacking gel and a wide pH difference between the gel buffer and the electrophoresis buffer at the beginning of the electrophoresis resulted in reduced band width and increasing resolution in silver-stained polyacrylamide gels. The importance of sharp DNA fragments is especially emphasized when analyzing multi-allelic DNA loci, that exhibit alleles differing from only few bp to few dozen bp in length, such as variable number of tandem repeat (VNTR) or short tandem repeat (STR) loci.

Asunto(s)

Alelos , ADN/genética , Electroforesis en Gel de Poliacrilamida/métodos , Amplificación de Genes/genética , Reacción en Cadena de la Polimerasa/métodos , Secuencias Repetitivas de Ácidos Nucleicos , Apolipoproteínas B/genética , Mapeo Cromosómico , Marcadores Genéticos/genética , Humanos

RESUMEN

Allele and genotype frequencies for the highly polymorphic D1S80 locus were determined in a Finnish population sample by using PCR followed by high-resolution PAGE and silver staining, a procedure called the amplified-fragment-length polymorphism (Amp-FLP) technique. In 140 unrelated Finnish individuals 15 alleles and 43 phenotypes were observed. The D1S80 locus demonstrated a heterozygosity of .77, and the power of discrimination was .92 in this sample representing a genetically isolated Finnish population. The distribution of observed genotypes conformed to Hardy-Weinberg expectations. In 36 mother-child pairs Mendelian inheritance for the alleles at the D1S80 locus could be demonstrated in all cases, and no mutations were observed. The usefulness of the D1S80 locus for forensic casework was assessed by using Amp-FLP analysis of the D1S80 locus in 36 forensic cases including 18 rapes, 14 homicides, and 4 other violent crimes. In most cases valuable information was obtained using the Amp-FLP technique, and in no case was there indication of either false-positive or false-negative results.

Asunto(s)

Alelos , Dermatoglifia del ADN , ADN/química , Genoma Humano , Población Blanca , Secuencia de Bases , Finlandia/etnología , Genotipo , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Estados Unidos/etnología

RESUMEN

The present study evaluates the usefulness of a PCR-based method for routine paternity testing in 35 paternity cases. This identification method which is based on amplification of three hypervariable genetic loci, apoB, D1S80 and HLA-DQ alpha, is compared, with regard to reliability and technical feasibility, to the conventional identification methods based on protein polymorphisms and to Southern blot hybridizations with multi- and single locus probes. Data obtained by PCR-amplification of these three loci resulted in paternity indices (56.1, geometric mean value) which are at the same level as the corresponding values derived from standard genetic blood group markers (42.7). The geometric mean value of the paternity indices obtained by Southern blot hybridization using three single locus probes (190.6) was more informative, and the most informative analysis proved to be Southern blot hybridization with multilocus probes. The technical feasibility and the reproducibility of the PCR-based analysis is, however, overwhelming, and if several highly polymorphic loci are amplified, the resolving power of PCR-analysis is similar to that obtained using multilocus probes.

Asunto(s)

ADN/genética , Paternidad , Reacción en Cadena de la Polimerasa/métodos , Tipificación y Pruebas Cruzadas Sanguíneas/métodos , Southern Blotting/métodos , Dermatoglifia del ADN/métodos , Variación Genética/genética , Humanos , Masculino , Secuencias Repetitivas de Ácidos Nucleicos/genética

RESUMEN

We have identified a hitherto unknown genetic polymorphism of apolipoprotein A-IV (apoA-IV). The molecular basis for this polymorphism is an A to G substitution at nucleotide 1687 resulting in an Asn to Ser change of amino acid 127. The frequencies of the two apoA-IV alleles (designated apoA-IV127Asn and apoA-IV127Ser), determined by Hin c II restriction analysis of PCR amplified exon three of the apoA-IV gene, were 0.788 and 0.212, respectively, in a Finnish population sample. Allele frequencies of another polymorphism due to a Thr to Ser substitution at amino acid 347 were determined using Hinf I restriction analysis. The allele frequencies were 0.823 for apoA-IV347Thr and 0.177 for apoA-IV347Ser. None of the apoA-IV polymorphisms (apoA-IV127:Asn----Ser, apoA-IV347:Thr----Ser and apoA-IV360:Gln----His) had any effect on plasma lipid and lipoprotein concentrations in cohorts of dyslipidemic men and in a population sample of normolipidemic controls. There was also no association between the history of previous myocardial infarction and any of the apoA-IV alleles.

Asunto(s)

Alelos , Apolipoproteínas A/genética , Asparagina , Serina , Secuencia de Aminoácidos , Apolipoproteínas A/análisis , Secuencia de Bases , Colesterol/sangre , Codón , Genotipo , Humanos , Hiperlipidemias/genética , Lipoproteínas/sangre , Masculino , Datos de Secuencia Molecular , Infarto del Miocardio/genética , Polimorfismo Genético , Treonina , Triglicéridos/sangre