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At least 649 records · Page 36Linked to original sources

PubMed1996

Early prenatal diagnosis of Fanconi anaemia in a twin pregnancy, using DNA analysis.

We present a case of prenatal diagnosis of Fanconi anaemia (FA) in a pair of twins at 14 weeks of gestation. The parents had previously had two children: a healthy boy and a boy with FA belonging to complementation group C (FAC). The FA patient is a compound heterozygote, carrying a 322delG and a IVS4+4A-->T mutation in the FAC gene. Prenatal DNA analysis showed that both fetuses were heterozygous for different mutations in the FAC gene. Both fetuses had normal male karyotypes. At 36 weeks the twins were born. They did not show congenital anomalies.

Adult↗

PubMed2004

Power-free poly(dimethylsiloxane) microfluidic devices for gold nanoparticle-based DNA analysis.

An extremely simple, power-free pumping method for poly(dimethylsiloxane)(PDMS) microfluidic devices is presented. By exploiting the high gas solubility of PDMS, the energy for the pumping is pre-stored in the degassed bulk PDMS, therefore no additional structures other than channels and reservoirs are required. In a Y-shaped microchannel with cross section of 100 microm width x 25 microm height, this method has provided flow rate of 0.5-2 nL s(-1), corresponding to linear velocity of 0.2-0.8 mm s(-1), with good reproducibility. As an application of the power-free pumping, gold nanoparticle-based DNA analysis, which does not rely on the cross-linking mechanism between nanoparticles, has been implemented in a microchannel with three inlets. Target 15mer DNA has been easily and unambiguously discriminated from its single-base substituted mutant. Instead of colorimetric detection in a conventional microtube, an alternative detection technique suitable for microdevices has been discovered-observation of deposition on the PDMS surfaces. The channel layout enabled two simultaneous DNA analyses at the two interfaces between the three laminar streams.

DNA↗

PubMedDate not supplied

Mutation-specific PCR: a rapid and inexpensive diagnostic method, as exemplified by mitochondrial DNA analysis in Leber's hereditary optic neuropathy.

Mitochondrial DNA analyses were carried out on 30 individuals with clinically documented or suspected Leber's hereditary optic neuropathy (LHON). Three methods based on the polymerase chain reaction (PCR) were compared, all three aiming at detecting the G/C to A/T mutation of base pair 11,778 causing LHON. Two methods included restriction analysis with either Sfa NI or Mae III, while the third one relied on allele-specific amplification (ASA), using a mutation-specific primer. The results were completely consistent, showing the presence of the mutation in 18 and its absence in 12 cases. From these results it is concluded, that mutation-specific PCR is the diagnostic method of choice, because it obviates the need for subsequent restriction analysis, thus being faster and more cost-efficient. The general applicability of ASA makes this strategy universally useful for detection of specific mutations in the diagnostic analysis of genetic disease, and for typing of genetic polymorphisms or other sequence variations due to single-base differences.

Base Sequence↗

PubMed1992

Characterization of Leptospira strains HY-1, HY-2, and HY-10 isolated in Korea by means of monoclonal antibodies and restriction endonuclease DNA analysis.

To analyze the characteristics of Leptospira strains HY-1, HY-2, and HY-10, which were isolated from patients with leptospirosis in Korea in 1985, 12 monoclonal antibodies (MAbs) against strain HY-1 and six MAbs against serovar lai strain 017 were produced, and their properties were determined by the microscopic agglutination test. Genetic relationships among the leptospires were determined by restriction endonuclease DNA analysis. Three MAbs reacted with all strains of serogroup Icterohaemorrhagiae, but did not react with any strains of the other 10 serogroups. All MAbs reacted with strains 017, HY-1, HY-2, and HY-10 at nearly identical titers. Two MAbs reacted only with these four strains. These four strains also had the same restriction endonuclease cleavage patterns. Based on these results, strains HY-1, HY-2, and HY-10 were identified as serovar lai, which is one of the common serovars in China. It is suggested that serovar lai is one of the prevalent serovars in Korea, and that the Mabs produced in this study are useful for the accurate and rapid identification of this serovar.

Agglutination Tests↗

PubMed2003

DNA fingerprinting of Ralstonia paucula by infrequent-restriction-site PCR and randomly amplified polymorphic DNA analysis.

Ralstonia paucula (formerly CDC group IV c-2) is an environmental organism that can cause serious human infections, occasionally clusters of nosocomial infections. In the present work, 26 strains of R. paucula (4 from the American Centers for Disease Control and Prevention collection, 10 from the Belgian Laboratorium voor Microbiologie [LMG] collection, and 12 French clinical isolates) were analyzed with infrequent-restriction-site PCR and randomly amplified polymorphic DNA analysis. Both techniques accurately distinguished between collection strains. Two close patterns obtained for all the French isolates suggested a clonal strain. Two LMG collection strains originating from human sources in the United States also showed patterns close to those of French isolates.

Child↗

PubMed1999

Effects of tamoxifen on steroid hormone receptors and hormone concentration and the results of DNA analysis by flow cytometry in endometrial carcinoma.

OBJECTIVES: Tamoxifen is a nonsteroidal triphenylethylene derivate with a predominant antiestrogen activity, used in the endocrine treatment of breast and endometrial cancer. It is not known which endometrial carcinomas will respond favorably to tamoxifen and which ones will not. The aim of this study was to find out whether tamoxifen has an effect on hormone steroid receptors, hormone concentration, DNA content, and proliferative activity in endometrial cancer and to correlate the tamoxifen-induced changes with pathologic parameters such as clinical stage, tumor differentiation, depth of invasion, and histologic type. METHODS: Thirty postmenopausal women with endometrial carcinoma were treated with 30 mg of tamoxifen daily for 7-10 days after curettage. Steroid hormone receptors (estrogen and progesterone receptors), levels of follicle-stimulating hormone, luteinizing hormone, prolactin, estradiol, progesterone, testosterone, dehydroepiandrosterone sulfate, sex hormone binding globulin, and DNA ploidy and proliferative activity were determined before and after therapy. The patients were also divided into favorable and unfavorable prognosis groups according to classical histological parameters. The patients in the favorable group consisted of patients with stage I disease, well and moderately differentiated tumors, favorable histologic type, and a depth of myometrial invasion of less than (1/3). The patients with only one of the unfavorable parameters (clinical stage II or III, poorly differentiated tumors, unfavorable histologic types, and deeper invasion of myometrium) were included in the unfavorable prognosis group. RESULTS: After the treatment, there was a net increase in the progesterone receptors and sex hormone binding globulin and a significant decrease in the estrogen receptors. The increase in progesterone receptors and decrease in estrogen receptors occurred in the patient group with favorable prognosis regarding histologic type, degree of differentiation, and clinical stage, but also in the unfavorable prognosis group regarding the depth of myometrial invasion. Statistically significant decrease in the follicle-stimulating hormone concentration was observed in the groups with favorable prognosis regarding histologic type, depth of myometrial invasion, and grade of differentiation. Concentration of sex hormone binding globulin was significantly increased in groups with favorable prognosis if histologic type and grade of differentiation were taken into account. On the other hand, there was a significant decrease in the concentration of luteinizing hormone in the group with unfavorable histologic type and also a decrease in progesterone concentration in patients with unfavorable prognosis regarding the grade of differentiation. There was no statistical significance either in the concentrations of other hormones measured or in the DNA analysis by flow cytometry. CONCLUSIONS: Our results revealed that tamoxifen can increase progesterone receptors and decrease estrogen receptors in endometrial cancer. The effect was most pronounced in tumors with favorable clinicopathologic parameters. We conclude that tamoxifen therapy can induce progesterone receptor synthesis even in tumors with low initial progesterone receptor levels, making such tumors potentially responsive to additional hormonal therapy with progesterone.

Adenocarcinoma↗

PubMed1991

An investigation of ring and dicentric chromosomes found in three Turner's syndrome patients using DNA analysis and in situ hybridisation with X and Y chromosome specific probes.

We have studied three patients with features of Turner's syndrome, two with a 45,X/46,X,r(?) and the third with a 45,X/46,X,dic?(Y) karyotype. Because Turner's syndrome patients with a mosaic karyotype containing a Y chromosome are known to have a high risk of developing gonadal tumours, we used DNA analysis and in situ hybridisation with X and Y specific probes to identify the chromosomal origin of the rings and dicentric chromosomes in the three index patients. Both ring chromosomes were shown to be of X origin, while the dicentric was composed of Y chromosome material. We discuss the importance of using a combination of molecular and cytogenetic analyses in such cases.

Adolescent↗

PubMed2000

A case of DNA analysis of ABO blood group variant allele A2.

The ABO phenotype of a bloodstain (B) on a knife that was used as a weapon in an attempted murder case was found to be different from that of the Peruvian victim's blood (AB). Serological analysis showed that the A-antigenicity was much weaker than B antigenicity, suggesting that the victim's phenotype was A(2)B or A(3)B. So, the ABO genotypes of the knife bloodstain and the victim's blood were determined by DNA analysis. The 261st G deletion, specific to the O(1) allele, was not detected in the specimens by restriction fragment length polymorphism analysis. Also, the 871st A, specific to the A(3) allele, was not found by the allele-specific amplification method. Amplified product length polymorphism and direct sequencing methods finally demonstrated that the typical B sequence was found in one allele and a single C deletion in the 1,059th-1,061st C stretch in the other allele, indicating that the ABO phenotype of the bloodstain and victim's blood were A(2)B.

Journal Article↗

PubMed2006

Routes to improving the reliability of low level DNA analysis using real-time PCR.

BACKGROUND: Accurate quantification of DNA using quantitative real-time PCR at low levels is increasingly important for clinical, environmental and forensic applications. At low concentration levels (here referring to under 100 target copies) DNA quantification is sensitive to losses during preparation, and suffers from appreciable valid non-detection rates for sampling reasons. This paper reports studies on a real-time quantitative PCR assay targeting a region of the human SRY gene over a concentration range of 0.5 to 1000 target copies. The effects of different sample preparation and calibration methods on quantitative accuracy were investigated. RESULTS: At very low target concentrations of 0.5-10 genome equivalents (g.e.) eliminating any replicates within each DNA standard concentration with no measurable signal (non-detects) compromised calibration. Improved calibration could be achieved by eliminating all calibration replicates for any calibration standard concentration with non-detects ('elimination by sample'). Test samples also showed positive bias if non-detects were removed prior to averaging; less biased results were obtained by converting to concentration, including non-detects as zero concentration, and averaging all values. Tube plastic proved to have a strongly significant effect on DNA quantitation at low levels (p = 1.8 x 10(-4)). At low concentrations (under 10 g.e.), results for assays prepared in standard plastic were reduced by about 50% compared to the low-retention plastic. Preparation solution (carrier DNA or stabiliser) was not found to have a significant effect in this study.Detection probabilities were calculated using logistic regression. Logistic regression over large concentration ranges proved sensitive to non-detected replicate reactions due to amplification failure at high concentrations; the effect could be reduced by regression against log (concentration) or, better, by eliminating invalid responses. CONCLUSION: Use of low-retention plastic tubes is advised for quantification of DNA solutions at levels below 100 g.e. For low-level calibration using linear least squares, it is better to eliminate the entire replicate group for any standard that shows non-detects reasonably attributable to sampling effects than to either eliminate non-detects or to assign arbitrary high Ct values. In calculating concentrations for low-level test samples with non-detects, concentrations should be calculated for each replicate, zero concentration assigned to non-detects, and all resulting concentration values averaged. Logistic regression is a useful method of estimating detection probability at low DNA concentrations.

Artifacts↗

PubMed2004

Human herpes virus-8 serology and DNA analysis in recipients of renal allografts showing Kaposi's sarcoma and their respective donors.

Kaposi's sarcoma (KS) developed among 11 of 416 renal allograft recipients transplanted between 1985 and 2000. Only 3 among 364 Caucasian recipients developed KS, while it affected 8 of 52 Black patients, all of whom had been born in African countries (P <.001). All patients had their immunosuppression reduced; two also received daunorubicin and one received electrotherapy. Three patients developed accelerated renal allograft dysfunction, probably due to the reduced immunosuppression. Remission of KS was observed in seven patients, while lesions stabilized or improved partially in the other four. After resuming dialysis 2 of 11 patients died; both were in KS remission. Human herpes virus-8 (HHV-8) serology and DNA analysis was evaluated in sera obtained from seven donors: all were negative. Conversely, among eight sera collected pretransplant from the nine living recipients, HHV-8 IgG was detected in six and DNA was present in one. HHV-8 IgG was expressed in all patients (9/9) at some point posttransplant; DNA was detected in three patients. Therefore, the robust ethnic predisposition to KS was associated with a high pretransplant prevalence of HHV-8 among African recipients. Although some seroconversions were detected posttransplant, there was no evidence for donor-to-recipient transmission.

Base Sequence↗

PubMed2005

DNA analysis of ingested tomato and pepper seeds.

Ingested food is one of the important types of forensic evidence obtained during a medicolegal autopsy. Many materials containing seeds pass through the human digestive system and are still recognizable; thus, they can be valuable for providing investigative leads. Currently, the identification of seeds relies on microscopic and morphologic examination. However this method sometimes can be problematic. For example, the microscopic appearance of the ingested tomato and pepper seeds is very similar; thus, it is not always easy to distinguish these seeds by comparing their physical characteristics. Tomato and pepper seeds were selected as a model system to assess the value of performing DNA analysis as an alternate and/or complimentary means of seed identification. Results of blind testing indicate that the deoxyribonucleic acid-amplified fragment length polymorphism (DNA-AFLP) results were able to discriminate between pepper and tomato seed samples after they passed through the digestive system.

Capsicum↗

PubMed1984

Hemoglobin Miyada: DNA analysis of the anti-Lepore beta delta fusion gene.

Hemoglobin Miyada, an anti-Lepore hemoglobin, represents the protein product of a nonhomologous crossover between beta and delta genes. The mutant globin is beta-like from the N terminus to amino acid 12, and delta-like from amino acid 22 through the C terminus, thus predicting a crossover site in the first coding region of the gene. DNA analysis, using multiple restriction endonucleases and hybridization to delta and beta globin gene-specific probes, confirms that the beta delta hybrid gene 1) is located on a single chromosomal fragment between normal delta and beta genes, and 2) has 5' beta promoter sequences, delta IVS 1 and 2, and 3' coding and flanking sequences.

Base Sequence↗

PubMedDate not supplied

Flow cytometric DNA analysis demonstrates contralateral testicular deterioration in experimental unilateral testicular torsion of prepubertal rats.

It has been postulated that unilateral testicular torsion causes damage to the contralateral testis and reduces fertility. However, in animal studies such an effect has not been fully proven by histopathologic examination or other conventional assays of spermatogenesis. We investigated the effect of unilateral testicular torsion on contralateral spermatogenesis in prepubertal rats using quantitative flow cytometric DNA analysis. Male rats were divided into three groups which underwent sham-operation, simple hemiorchiectomy or unilateral testicular torsion. Five weeks after these operations, fertility and spermatogenesis by flow cytometry were evaluated. No significant differences were observed in body weight, contralateral testicular weight or serum testosterone concentration among the three experimental groups. In the torsion group, mean seminiferous tubular diameter, number of foetuses, fertility rate and percentage of haploid cells were all significantly decreased compared to the other two groups. These results suggest that unilateral testicular torsion causes damage to the contralateral testis and consequently can reduce the future fertility of prepubertal rats.

Animals↗

PubMed1995

DNA analysis of dental pulp to link incinerated remains of homicide victim to crime scene.

Teeth endure postmortem degradation and extreme changes in ambient temperature and pressure better than most human tissues. This ability to resist deterioration allows the teeth to be studied as a method of establishing the identity of a decedent. Additionally, dental hard tissues, and in some instances soft tissues, may provide investigators with other sources of forensic data. In this case, a female homicide victim was transported to a location where her remains were burned. The high temperatures of a gasoline fire effectively incinerated the body precluding deoxyribonucleic acid (DNA) analysis from conventional sites. However, most of the teeth survived the conflagration. They were used to identify the victim. Additionally, the dental pulps were found to be an excellent source of high molecular weight genomic DNA. This proved to be an effective method to link the victim's body to biological evidence recovered from the site of the murder.

Adult↗

PubMed2001

Direct DNA analysis in a new Italian carrier of Hb-Belfast: beta 15 (A12) Trp-->Arg.

A young woman aged 21 was found to be a new carrier of Hb-Belfast: beta 15 (A 12) Trp-->Arg, and the characteristics of her hemoglobinopathy were not different from those of the four cases so far described: mild hemolysis with molecular instability of the abnormal Hb, red cells inclusion bodies, and slight alterations of some functional parameters of whole blood. On this occasion, direct DNA analysis indicated the genomic nucleotide replacement of the disease: TGG-AGG. This was inherited by the mother, originating from Bari (Apulia).

Adult↗

PubMed1990

[DNA analysis of HLA class II and III genes in sudden infant death (SIDS)].

In 39 cases of SID we studied the polymorphism of HLA-class II genes DRB and DQB and HLA-class III genes C4 (C4A and C4B, genes encoding the fourth complement component) and 21-hydroxylase (21OH-A and 21OH-B, genes encoding the enzyme 21-hydroxylase of the cortisol pathway) by DNA analysis. This study was performed in cooperation with the Institute of Legal Medicine, University of Munich. Compared to healthy controls the percentage of C4B gene deletions, C4B gene duplications (C4B "short") and 21OH-A gene deletions is increased in SIDS. The deviations are statistically not significant. The analysis of the HLA-DR alleles revealed a significant decrease of HLA-DR2 in SIDS compared to controls (p = 0.0016, pc = 0.016). The results of this study indicate a possible role of C4B/21OH-A gene defects as a risk factor in a subgroup of SID cases. This hypothesis has to be confirmed by studying further cases.

Chromosome Aberrations↗

PubMed1993

Increased echogenicity in the fetal abdomen: use of DNA analysis to establish a diagnosis of cystic fibrosis.

The sonographic finding of increased echogenicity within the fetal abdomen presents a diagnostic dilemma, with a differential diagnosis ranging from normal variation to CF. We report the diagnostic evaluation of four cases, two of which were found to be the result of CF. On the basis of this experience, we believe that persistence of an echogenic bowel pattern, especially with bowel dilation, after 20 weeks' gestation should prompt an evaluation for CF. Using DNA analysis, approximately 75% of the cases involving CF can be detected with noninvasive studies of the parents, and confirmation by amniocentesis is performed only in those cases in which both parents are carriers of known mutations.

Abdomen↗

PubMed1997

Pulsed-field gel electrophoresis is more discriminating than multilocus enzyme electrophoresis and random amplified polymorphic DNA analysis for typing pyogenic streptococci.

The SmaI restriction endonuclease digestion patterns of chromosomal DNAs from 99 pyogenic streptococci belonging to Lancefield group A (41 Streptococcus pyogenes), group C (seven S.dysgalactiae, 11 "S. equisimilis", three S. equi, eight S. zooepidemicus) and group G (25 human group G Streptococcus, four S. canis) were analyzed by pulsed-field gel electrophoresis (PFGE), and the results were compared with those previously obtained by multilocus enzyme electrophoresis (MLEE) and random amplified polymorphic DNA analysis (RAPD). PFGE revealed 93 distinct types among the 99 strains, and no patterns were common to strains of different species. The discriminatory power of PFGE was greater than that of MLEE and RAPD for groups A and G streptococci. The polymorphism among group C streptococci was similar with the three techniques. PFGE is, therefore, the most efficacious method for epidemiological typing of pyogenic streptococci.

DNA, Bacterial↗