Forensic nurse, crime experts, urge U.S. Senators to overhaul DNA analysis system.
Explore the source record for details and available documents.
DNA Fingerprinting↗
SEARCH · Search PubMed
Results for “DNA analysis”
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
Flow cytometric DNA analysis of neuroblastoma--clinical significance of DNA tetraploidy.
Explore the source record for details and available documents.
Child, Preschool↗
DNA analysis with multiplex microarray-enhanced PCR.
We have developed a highly sensitive method for DNA analysis on 3D gel element microarrays, a technique we call multiplex microarray-enhanced PCR (MME-PCR). Two amplification strategies are carried out simultaneously in the reaction chamber: on or within gel elements, and in bulk solution over the gel element array. MME-PCR is initiated by multiple complex primers containing gene-specific, forward and reverse, sequences appended to the 3' end of a universal amplification primer. The complex primer pair is covalently tethered through its 5' end to the polyacryl- amide backbone. In the bulk solution above the gel element array, a single pair of unattached universal primers simultaneously directs pseudo-monoplex PCR of all targets according to normal solution-phase PCR. The presence of a single universal PCR primer pair in solution accelerates amplification within gel elements and eliminates the problem of primer interference that is common to conventional multiplex PCR. We show 10(6)-fold amplification of targeted DNA after 50 cycles with average amplification efficiency 1.34 per cycle, and demonstrate specific on-chip amplification of six genes in Bacillus subtilis. All six genes were detected at 4.5 pg of bacterial genomic DNA (equivalent to 10(3) genomes) in 60 independent amplification reactions performed simultaneously in single reaction chamber.
DNA↗
Flow cytometric DNA analysis of parathyroid glands. Relationship between nuclear DNA and pathologic classifications.
Nuclear DNA contents of 95 paraffin-embedded parathyroid glands (2 carcinomas, 56 adenomas, 10 primary and 17 secondary chief cell hyperplasias, and 10 normal glands) were determined by flow cytometric analysis. All normal parathyroid glands and secondary hyperplasias, 80% of the primary hyperplasias, and 73% of the adenomas had diploid DNA patterns, with 15% or less tetraploid cells. Twenty-one percent of the adenomas, 29% of the primary hyperplasias, and all carcinomas had diploid and tetraploid DNA distribution patterns, with greater than 15% of the cells in the tetraploid region. One of the carcinomas (Figure 4B, Case 2) had an additional near-triploid aneuploid peak. Three of the adenomas (5.4%) had near-triploid aneuploid patterns. One of the patients with carcinoma (Figure 4A, Case 1) died, 32 months after initial diagnosis, of disease-related causes. The remaining patient with carcinoma (Case 2) had a 47-month disease-free interval. All of the patients with hyperplastic and adenomatous glands are free of disease, after a mean follow-up interval of 25 months. This study indicated that flow cytometric analysis of nuclear DNA content does not complement conventional pathologic methods in distinguishing between parathyroid gland chief cell hyperplasia, adenoma, or carcinoma; however, it did suggest the possibility that parathyroid adenomas and primary chief cell hyperplasias may contain a subset of tumors that could manifest biologic malignancy if allowed to progress untreated.
Adolescent↗
Immunocytochemical determination of estrogen and progesterone receptors and flow cytometric DNA analysis of breast cancer on fine needle aspirates.
A prospective study was performed on a series of 42 patients with breast carcinoma. Immunocytochemical determination of receptors for estrogens (ER) and progesterone (PR) and flow cytometric DNA analysis (DNA-ploidy and the S-phase fraction) were performed on material obtained from fine-needle aspirates (FNA). The patients underwent subsequent surgery, and the same parameters were determined on the surgical samples. The correlation between the biochemical and the immunocytochemical methods gave for ER, r = 0.853, p < 10 (-6) and for PR, r = 0.830, p < 1.6 (-6). In addition, comparisons of FNA's with the surgical samples showed significant relationships for the DNA index (r = 0.96, p < 0.000001) and for the S-phase fraction (r = 0.88, p < 0.000001).
Adult↗
Prospective flow cytometric DNA analysis of hepatocellular carcinoma specimens collected by ultrasound-guided fine needle aspiration.
BACKGROUND: The survival of 52 patients with hepatocellular carcinoma (HCC) seen during the last 4 years was analyzed prospectively on the basis of disease stage and nuclear DNA content. METHODS: Ploidy was measured by flow cytometry (FCM). Cells for cytologic diagnosis and FCM were collected by ultrasound-guided fine needle aspiration. RESULTS: DNA aneuploidy, which was detected in 62% of the patients, did not correlate with clinicopathologic features, except in the sonographic aspect (P = 0.03). However, ploidy correlated significantly with survival; the survival times for patients with an aneuploid DNA index were significantly shorter than for those with a diploid index (P = 0.02). In a Cox multivariate analysis, DNA content was prognostically significant, as were the grade of cirrhosis severity and the echographic aspect. CONCLUSIONS: In addition to the clinicopathologic features observed, FCM DNA analysis of ultrasound-guided fine needle aspirates from HCC is a simple and valid method for estimating a prognosis of these patients.
Aged↗
DNA analysis as a tool to confirm the diagnosis of asymptomatic hereditary neuropathy with liability to pressure palsies (HNPP) with further evidence for the occurrence of de novo mutations.
We performed DNA analysis in four families with hereditary neuropathy with liability to pressure palsy (HNPP). An interstitial deletion of the 17 p11.2 region was found in typically affected patients as well as in as yet asymptomatic patients. The opportunity for an individual genotyping permitted to ascertain a de novo deletion in one clinically affected case with no relevant familial history. DNA analysis thus becomes the most sensitive tool in diagnosing HNPP, since potentially affected patients may lack either informative familial history, or clinical symptoms or even suggestive EMG or histopathological data (tomaculas).
Adolescent↗
Rapid discrimination of Mycobacterium avium strains from AIDS patients by randomly amplified polymorphic DNA analysis.
A randomly amplified polymorphic DNA (RAPD) analysis was performed for the molecular typing of Mycobacterium avium strains. This method was applied to epidemiologically unrelated M. avium strains isolated from the blood of 10 different AIDS patients and to strains that were considered epidemiologically related, as they had been isolated from the same patient but from different body locations (4 patients, 10 strains). Three oligonucleotide primers among the six tested were found to generate RAPD profiles with DNA from all M. avium strains and to successfully type them. This method for the typing of M. avium strains is rapid and easy to perform.
AIDS-Related Opportunistic Infections↗
Allogeneic cultured epidermal grafts heal chronic ulcers although they do not remain as proved by DNA analysis.
To investigate whether allogeneic cultured keratinocytes are rejected or not, and to find out how beneficial their effect on wound healing could be, patients with chronic ulcers were grafted with allogeneic cultured human keratinocytes. In order to examine the epidermal origin of the healed wound, DNA analysis was performed and compared to donor and recipient blood-cell DNA. Healing was observed in 84% of the grafted ulcers by granulation tissue stimulation and would edge effect. In little time 60% of the grafted chronic ulcers healed completely. Although no rejection was observed, DNA analysis revealed that the grafted allogeneic keratinocytes were finally replaced by the patient's own epidermis. This study confirmed that cultured allogeneic keratinocytes that have been grafted on ulcers, play an important role in the wound healing process.
Adult↗
Ultrafast DNA analysis by capillary electrophoresis/laser-induced fluorescence detection.
The limits of ultrafast DNA analysis by CE were determined by investigating the influence of the effective capillary length and the electric field strength on the analysis time for a given peak resolution (10 bp). In accordance with theory, the use of a fast ramp power supply for narrow plug electrokinetic injection was found to be essential to minimize the extra column effects on peak dispersion. Two major column dispersion factors, longitudinal diffusion and thermal dispersion, were determined experimentally, as well as the influence of the electric field strength on the electrophoretic mobilities and diffusion coefficients of DNA. It was found that higher field strengths can be applied with lower thermal dispersion than predicted by classical CE models. This was attributed to the faster mass transport in the radial direction due to field-induced DNA orientation. Short capillaries (approximately 3-7 cm effective length) and moderate to high electric field strengths (approximately 600-800 V/cm) were used to perform a series of fast DNA separations. The dsDNA fragment standards phiX174/HaeIII and pBR322/HaeIII were separated within 30 s. The possibility for fast mutation detection was demonstrated using constant denaturant capillary electrophoresis (CDCE) for the analysis of a single base mutation in mitochondrial DNA in 72 s. The potential for fast DNA sequencing was illustrated by separating 300 ssDNA fragments within 180 s.
DNA↗
Prognostic significance of flow cytometric DNA analysis in node-negative breast cancer patients.
Flow cytometric DNA analysis using paraffin-embedded tumor blocks was done retrospectively on 155 node-negative breast cancers. The median duration of follow-up in patients still alive at the time of analysis was 10 years. Tumor aneuploidy was correlated significantly with increased tumor size (P = 0.003) and higher tumor grade (P less than 0.001). No significant correlation between tumor ploidy and patient age was found. Patients with diploid tumors had a significantly improved relapse-free and overall survival compared with patients with aneuploid tumors (P = 0.0001). In a Cox multivariate model with parameters including ploidy, histologic grade, tumor size, and patient age, ploidy (P = 0.02) and tumor size (P = 0.05) emerged as significant independent predictors of overall survival. Only ploidy was independently significant in the analysis of relapse-free survival. In conclusion, the current study indicates that flow cytometric measurement of DNA ploidy is a powerful prognostic indicator in node-negative breast cancer patients.
Breast Neoplasms↗
Mitochondrial DNA analysis of Exophiala moniliae.
Mitochondrial DNA (mtDNA) analysis with restriction enzymes, Hae III, Hind III and Msp I was performed in 17 Exophiala moniliae strains. The results were as follows: (1) E. moniliae could be classified into 10 types based on restriction patterns, (2) E. moniliae is suggested to be a complex organism because of extensive mtDNA polymorphism among strains like E. jeanselmei and (3) two types of E. moniliae are identical with two types of E. jeanselmei. These results suggest that E. moniliae is not genetically defined from E. jeanselmei and the taxonomical status of E. moniliae requires reevaluation.
DNA Restriction Enzymes↗
Screening for the FV:Q506 mutation--evaluation of thirteen plasma-based methods for their diagnostic efficacy in comparison with DNA analysis.
The factor V (FV) mutation Q506 that causes resistance to activated protein C (APC) is the genetic defect associated most frequently with venous thrombosis. The laboratory diagnosis can be made by DNA analysis or by clotting tests that measure the degree of prolongation of plasma clotting time upon addition of APC. Home-made and commercial methods are available but no comparative evaluation of their diagnostic efficacy has so far been reported. Eighty frozen coded plasma samples from carriers and non-carriers of the FV:Q506 mutation, diagnosed by DNA analysis, were sent to 8 experienced laboratories that were asked to analyze these samples in blind with their own APC resistance tests. The APTT methods were highly variable in their capacity to discriminate between carriers and non-carriers but this capacity increased dramatically when samples were diluted with FV-deficient plasma before analysis, bringing the sensitivity and specificity of these tests to 100%. The best discrimination was obtained with methods in which fibrin formation is triggered by the addition of activated factor X or Russell viper venom. In conclusion, this study provides evidence that some coagulation tests are able to distinguish carriers of the FV:Q506 mutation from non-carriers as well as the DNA test. They are inexpensive and easy to perform. Their use in large-scale clinical trials should be of help to determine the medical and economic benefits of screening healthy individuals for the mutation before they are exposed to such risk factors for venous thrombosis as surgery, pregnancy and oral contraceptives.
Anticoagulants↗
[Forensic DNA-analysis in pathology].
The principles of modern forensic DNA-analysis using the amplification of so called STR-polymorphisms are explained. Four cases from practise that required the collaboration of forensic molecular biology and pathology and which were analysed by them are presented and discussed. Similarities and variations in the work of both "disciplines" are presented and suggestions for the storage of samples formulated.
Adult↗
Flow DNA analysis in the characterization of carcinoma of the renal pelvis and ureter.
A prospective study comprised operative specimens from 11 patients with transitional cell carcinoma of the renal pelvis or ureter. DNA analysis of the primary tumor and of multiple biopsy specimens from preselected sites of the surrounding urothelium was performed with flow cytometry. All Grade 3 tumors and 50% of the Grade 2 tumors were aneuploid, and the remainder were Grade 2 and diploid. All invasive tumors were aneuploid. Carcinoma in situ was found in some of the preselected biopsy specimens, all of which were aneuploid, from two patients. Close correlation thus was observed between aneuploidy and tumor invasiveness, whereas diploidy was seen only in noninvasive tumors with lower malignancy grade. Aneuploidy was also associated with increased risk of carcinoma in situ. The study indicated that DNA analysis may be useful for defining the malignant potential of urothelial tumors of the upper urinary tract more fully than conventional grading and staging permit.
Adult↗
DNA analysis to aid in the diagnosis of chronic myeloproliferative disorders.
Diagnosing chronic myeloproliferative disorders (CMPD) can be difficult because of overlap and possible transitions between the different conditions and their similarity to reactive myeloproliferations. DNA analysis was applied to improve differentiation of CMPDs. All subtypes of CMPD analyzed, including chronic myeloid leukemia, agnogenic myeloid metaplasia, polycythemia vera, and essential thrombocythemia, had in common that granulocytes and bone marrow cells were clonal in origin, as shown by X chromosome-linked DNA polymorphism in conjunction with methylation patterns (n = 32). Reactive myeloproliferations, by contrast, showed polyclonal inactivation patterns. Clonality could not distinguish CMPD from cases of myelodysplastic syndrome because the latter (n = 7) also exhibited clonal hematopoiesis. Because of their clonal origin, peripheral granulocytes were used in all cases (n = 201) to detect bcr gene rearrangement. Despite possible morphologic overlap between different types of CMPD, bcr gene rearrangement was specific for chronic myeloid leukemia and could be applied to differentiate chronic myeloid leukemia from other CMPDs in cases of equivocal morphologic diagnosis. Chronic myeloproliferative disorders represent clonal hemopoietic diseases that probably have specific underlying genetic defects. Thus DNA analysis can aid substantially in the differential diagnosis of CMPD.
Adolescent↗
Quantitative DNA analysis in breast carcinomas: a comparison between image analysis and flow cytometry.
We compared the results of quantitative DNA analysis of fresh tumor tissue from 50 invasive breast carcinomas by image analysis and flow cytometry. For image analysis, Feulgen-stained slides of tumor imprints and of disaggregated tumor cytospin preparations were evaluated with the CAS-200 image analyzer. For flow cytometry, propidium iodide-stained disaggregated tumor cells were analyzed with the Coulter EPICS-C flow cytometer. The two methods yielded comparable results. The DNA indices obtained by the two methods showed close correlation by linear regression analysis (r = 0.86, P less than 0.001). There were 26 diploid (52%) and 24 nondiploid (48%) carcinomas. The ploidy pattern between the two methods showed agreement in 41 carcinomas (82%) and discordance in two (4%). Three tumors (6%) were equivocal by flow cytometry and four (8%) by image analysis. The equivocal cases presented potential sources of error in the evaluation of histograms in the near-tetraploid region by flow cytometry and in the near-diploid region by image analysis. Image analysis required smaller tissue samples and permitted direct visualization and selection of tumor cells. It also detected more tetraploid carcinomas. In contrast, flow cytometry analyzed larger cell samples and provided histograms with better resolution. It more readily detected the presence of multiple aneuploid peaks and also the presence of aneuploid peaks in the near-diploid range. The presence of aneuploidy was significantly related to the loss of hormone receptor expression, high mitotic rate, and high histologic and nuclear grades. Our study indicates that image analysis and flow cytometry provide comparable results in a majority of breast carcinomas.(ABSTRACT TRUNCATED AT 250 WORDS)
Breast Neoplasms↗
Clinical applications of plasma Epstein-Barr virus DNA analysis and protocols for the quantitative analysis of the size of circulating Epstein-Barr virus DNA.
Nasopharyngeal carcinoma (NPC) is one of the most common cancers in Southern China. Epstein-Barr virus (EBV) infection is an important etiological factor of NPC. The fact that EBV genome is present in almost all NPC tissues renders it an ideal tumor marker for NPC. To date, quantitative analysis of plasma EBV DNA has been shown to be clinically useful for the detection, monitoring, and prognostication of NPC. In addition, the molecular nature of circulating EBV DNA has recently been identified as that of free DNA fragments; it is not contained inside intact virions. By quantitative size analysis, it is further demonstrated that more than 80% of these DNA fragments are less than 180 bp in size. In this chapter, the clinical applications of plasma EBV DNA analysis and the protocols for the quantitative analysis of the size of circulating EBV DNA will be discussed.
Biomarkers, Tumor↗