SEARCH · Search PubMed

Results for “Rapid Genetic Testing”

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.

SourceResearch connectionFrom yearThrough year

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 721 records · Page 40Linked to original sources

PubMed1987

Recent developments in antiviral chemotherapy.

As can be seen from the preceding brief discussion, the prospects for new antiviral agents for the treatment of viral diseases ranging in severity from the common cold to AIDS appear promising. The latest advances in technologies such as crystallography, genetic engineering, and monoclonal antibodies are all being applied to the discovery of new ways to inhibit virus-specific processes. In addition, the development of diagnostic tests for viral diseases has proceeded at a rapid pace that should facilitate the proper use of the antivirals when they become available. The one lesson that has been learned in the antiviral field over the past decade is that virus-specific inhibitors do exist, and if an essential virus-specific process or protein can be identified, it is likely that a molecule can be found to inhibit or inactivate it.

Antiviral Agents↗

PubMed2001

Taxanes: the genetic toxicity of paclitaxel and docetaxel in somatic cells of Drosophila melanogaster.

In this study, the taxanes, paclitaxel and docetaxel were investigated for genotoxicity in the wing spot test of Drosophila melanogaster. These relatively new drugs are used in cancer therapy and show great promise in the treatment of a variety of cancers. Their major cellular target is the alpha,beta-tubulin dimer but, unlike other spindle poisons, they stabilize microtubules by a shift towards assembly, producing nonfunctional microtubule bundles. The Drosophila wing Somatic Mutation and Recombination Test (SMART) provides a rapid means to evaluate agents able to induce gene mutations and chromosome aberrations, as well as rearrangements related to mitotic recombination. We applied the standard version of SMART (with normal bioactivation) and a variant version with increased cytochrome P450-dependent biotransformation capacity. In the standard assay, docetaxel was found to be aneuploidogenic; this was effectively abolished by a high cytochrome P450-dependent detoxification capacity. This suggests, as previously reported, the involvement of this family of enzymes in the detoxification of docetaxel rather than in its activation. In contrast, paclitaxel was clearly non-genotoxic at the same (millimolar) concentrations as used for docetaxel in both crosses. The weak responsiveness of SMART assays to aneugenic compounds, the weaker ligand and assembly action of paclitaxel and the more rapid reversibility of the microtubules formed with this compound, may have caused the negative response observed in the present study.

Angiogenesis Inhibitors↗

PubMed1985

Temperature-sensitive mutations causing reversible paralysis in Caenorhabditis elegans.

A method has been developed for the isolation of temperature-dependent paralytic mutants of the nematode Caenorhabditis elegans, based on a screening procedure using short-time exposure to 30 degrees C. Of ten mutants isolated, eight lose their motilities between 30 degrees C and 33 degrees C without prominent changes in body forms. The other two strains that are mainly described in this report are accompanied by alterations in body forms. One mutation, cn101, is recessive and an allele of cha-1. The cn101 mutant shows reversible paralysis at 30 degrees, accompanied by a hypercontracted and coiled body form. At the restrictive temperature, the strain is resistant to all tested inhibitors of acetylcholinesterase (AChE). Another mutation, designated mah-2 (cn110), is a sex-linked semidominant that is mapped as 0.6 map units left of dpy-6. The cn110 mutant is rapidly paralyzed at the restrictive temperature and has a straight and rigid body form; the mutant rapidly recovers when the temperature is lowered. No disorganization of the muscle structure was detected by polarized light and electron microscopic inspection.

Alleles↗

PubMed2000

Cheap, accurate and rapid allele frequency estimation of single nucleotide polymorphisms by primer extension and DHPLC in DNA pools.

At present, the cost of genotyping single nucleotide polymorphisms (SNPs) in large numbers of subjects poses a formidable problem for molecular genetic approaches to complex diseases. We have tested the possibility of using primer extension and denaturing high performance liquid chromatography to estimate allele frequencies of SNPs in pooled DNA samples. Our data show that this method should allow the accurate estimation of absolute allele frequencies in pooled samples of DNA and also of the difference in allele frequency between different pooled DNA samples. This technique therefore offers an efficient and cheap method for genotyping SNPs in large case-control and family-based association samples.

Alleles↗

PubMed1997

Use of first nucleotide change technology to determine the frequency of factor V Leiden in a population of Australian blood donors.

Activated protein C resistance (APCR), the most common risk factor for venous thrombosis, is the result of a G to A base substitution at nucleotide 1691 (R506Q) in the factor V gene. Current techniques to detect the factor V Leiden mutation, such as determination of restriction length polymorphisms, do not have the capacity to screen large numbers of samples in a rapid, cost-effective test. The aim of this study was to apply the first nucleotide change (FNC) technology, to the detection of the factor V Leiden mutation. After preliminary amplification of genomic DNA by polymerase chain reaction (PCR), an allele-specific primer was hybridised to the PCR product and extended using fluorescent terminating dideoxynucleotides which were detected by colorimetric assay. Using this ELISA-based assay, the prevalence of the factor V Leiden mutation was determined in an Australian blood donor population (n = 500). A total of 18 heterozygotes were identified (3.6%) and all of these were confirmed with conventional MnlI restriction digest. No homozygotes for the variant allele were detected. We conclude from this study that the frequency of 3.6% is compatible with others published for Caucasian populations. In addition, the FNC technology shows promise as the basis for a rapid, automated DNA based test for factor V Leiden.

Adenine↗

PubMed2000

Drug-resistant tuberculosis: what do we do now?

Drug-resistant tuberculosis (TB) represents a threat to TB control programmes. Erratic and inappropriate use of currently available medications, HIV-TB co-infection, and concern about transmission of drug-resistant strains in the general population all contribute to a worrying picture. What do we do now? In the last few years, there has been considerable progress in the understanding of mechanisms of action and resistance to antituberculosis agents, and in establishing the value of directly observed therapy in preventing treatment failure. However, a limited effort has been devoted to the development of new active compounds or of rapid diagnostic tests, and their relevance to global tuberculosis control has been questioned.

Antitubercular Agents↗

PubMed2004

Detection of rifampin-resistant Mycobacterium tuberculosis strains by using a specialized oligonucleotide microarray.

DNA microarray represents one of the major advances in diagnostic sequencing of polymerase chain reaction (PCR) products. Until now, arrays have been relatively expensive, complex to perform, and difficult to interpret, limiting their wide application in the clinical laboratory. A moderate-density oligonucleotide microarray that can rapidly identify Mycobacterium tuberculosis rifampin-resistant strains was developed. The method is based on the detection of point mutations and other rearrangements in the rpoB gene region determining rifampin resistance. Rifampin resistance was determined by hybridizing fluorescently labeled, amplified genetic material generated from bacterial colonies to the array. Fifty-three rifampin-resistant M. tuberculosis and 15 rifampin-susceptible M. tuberculosis were tested and results were concordant with those based on culture drug susceptibility testing and sequencing. Rifampin-resistant clinical isolates were detected in as little as 1.5 hours after PCR amplification with visual results. It is demonstrated that oligonucleotide microarray is an efficient, specialized technique to implement and can be used as a rapid method for detecting rifampin resistance to complement standard culture-based method.

Antibiotics, Antitubercular↗

PubMed2005

Isolation by distance, web service.

BACKGROUND: The population genetic pattern known as "isolation by distance" results from spatially limited gene flow and is a commonly observed phenomenon in natural populations. However, few software programs exist for estimating the degree of isolation by distance among populations, and they tend not to be user-friendly. RESULTS: We have created Isolation by Distance Web Service (IBDWS) a user-friendly web interface for determining patterns of isolation by distance. Using this site, population geneticists can perform a variety of powerful statistical tests including Mantel tests, Reduced Major Axis (RMA) regression analysis, as well as calculate FST between all pairs of populations and perform basic summary statistics (e.g., heterozygosity). All statistical results, including publication-quality scatter plots in Postscript format, are returned rapidly to the user and can be easily downloaded. CONCLUSION: IBDWS population genetics analysis software is hosted at http://phage.sdsu.edu/~jensen/ and documentation is available at http://www.bio.sdsu.edu/pub/andy/IBD.html. The source code has been made available on Source Forge at http://sourceforge.net/projects/ibdws/.

Computer Graphics↗

PubMed2002

Evaluating porcine RBC and platelet alpha-galactosyl expression.

BACKGROUND: Naturally occurring human xenoreactive antibodies bind and agglutinate porcine RBCs. STUDY DESIGN AND METHODS: To determine if xenoantigen expression on RBCs of individual pigs of different breeds and blood groups is variable, and if it correlates with platelet (PLT) expression, we measured adsorption of affinity-purified antibodies to alpha-galactosyl (alphaGal) by RBCs or PLTs from 22 pigs representing four breeds. Hemagglutination of RBCs from these pigs was also performed with pools of human group OAB, A, B, and AB sera, as well as with anti-alphaGal-depleted pooled OAB human serum. RESULTS: There was significant variation in alphaGal expression on RBCs and PLTs among pigs. PLT alphaGal expression did not correlate with RBC alphaGal. RBCs from all pigs were agglutinated by pooled group O, AB, A, or B sera, whereas titers were reduced by 87 percent with anti-alphaGal-depleted serum and by 82 percent with AB sera from two volunteers. Agglutination titers were higher against RBCs from the five highest RBC alphaGal expressers compared with those from the five lowest RBC alphaGal expressers (92 +/- 12 vs. >160, p < 0.05, where 160 was the maximum dilution tested). CONCLUSION: Hemagglutination is a feasible alternative for rapid identification of pigs with RBCs carrying less alphaGal.

ABO Blood-Group System↗

PubMed2000

Microbial ecology and treatment of Helicobacter pylori infections: review.

The aims of the present study were to investigate the ecological disturbances caused by four different anti-H. pylori regimens, to compare different methods for diagnosing H. pylori, and to study the genetic variability of H. pylori. The patients included in the study were all treated at the Center of Gastroenterology, Huddinge University Hospital, Karolinska Institute. All patients were H. pylori-positive before entering the study, confirmed by rapid urease test, histology, culture and urea breath test or PCR. Treatment regimens included in the study were omeprazole alone (OP), in combination with amoxicillin (OA), in combination with amoxicillin and metronidazole (OAM) and in combination with clarithromycin and metronidazole (OCM). Samples from the mouth (saliva and dental plaque), stomach (biopsies from the gastric mucosa in the corpus and in the antrum) and the intestine (feces) were collected before, during and after treatment. The oral microflora was challenged by the three treatment regimens including antimicrobial agents, with the emergence of resistant streptococci and staphylococci in the OCM group. Bacterial strains in the gastric mucosa increased in numbers during treatment in all treatment groups, probably due to the pH rise, which provides a better environment for the commensal microflora. This overgrowth was especially pronounced during treatment with omeprazole alone (OP), possibly due to the fact that a concomitant suppression exerted by the antimicrobial agents occurred in the other treatment groups. H. pylori was, on the other hand, suppressed during treatment in all treatment groups, possibly due to a direct effect of omeprazole and to the colonization resistance expressed by the normal microflora. An emergence of resistant commensal strains in the gastric mucosa was seen in the OCM and the OAM groups. The intestinal microflora was most altered in the OAM and the OCM groups, with persistent disturbances in the OCM group 4 weeks after treatment. The frequency of resistant Enterococcus spp. (OCM), Enterobacteriaceae spp. (OA and OAM) and Bacteroides spp. (OCM) was increased during and after treatment. Different detection methods for H. pylori were compared and PCR was shown to have higher sensitivity than other routine diagnostic tests. The patients in the present study seemed to be colonized with a single strain of H. pylori. Treatment failures in patients treated with OAM were caused by recrudescence. These four patients with relapsing H. pylori infection, were shown to be reinfected with the original H. pylori strain, indicating that H. pylori escapes treatment by a thus far unknown mechanism.

Adenocarcinoma↗

PubMed2004

FGF-7 expression enhances the performance of bioengineered skin.

To improve the performance of bioengineered skin, we used a recombinant retrovirus encoding FGF-7 to modify diploid human keratinocytes genetically. Control or FGF-7-expressing keratinocytes were seeded onto acellular human dermis to form bioengineered skin. Gene-modified skin secreted significant levels of FGF-7 and formed a thicker and hyperproliferative epidermis with about four times the number of cells per square centimeter. Secretion of an endogenous trophic factor, VEGF, was increased approximately 5-fold. Migration of FGF-7-expressing keratinocytes was stimulated as was the self-healing of bioengineered skin expressing FGF-7. When tested in a bacterial infection model, the antimicrobial properties of FGF-7-expressing skin were increased >500-fold against both gram-negative and gram-positive bacteria. After transplantation to full-thickness wounds on athymic mice, skin expressing FGF-7 was revascularized more rapidly. These results demonstrate that genetic modification can be used to enhance performance and that expression of FGF-7 augments several properties important to the wound-healing properties of bioengineered skin.

Animals↗

PubMed2003

Identification of three genetic risk factors for venous thrombosis using a multiplex allele-specific PCR assay: comparison of conventional and new alternative methods for the preparation of DNA from clinical samples.

The demand for thrombophilia testing at the molecular level is increasing and with it the need for a simple and rapid and cost-saving procedure for the preparation of genomic DNA from whole blood samples. The aim of this paper is to compare the efficiency of two conventional commercial procedures (Genomic, Eurobio-Labtek, and Nucleospin, Macherey-Nagel) and two our alternative approaches (microwave irradiation and resin-binding method) for extraction DNA and their suitability and convenience for multiple sample preparation for simultaneous identification of the factor V Leiden, prothrombin 20210 and methylene tetrahydrofolate reductase (MTHFR) 677 variants by multiplex allele specific amplification (ASA-PCR). We have found that chemical-based kit (Genomic) produced higher DNA recovery (mean recovery 40 +/- 4.2 microg/ml; A260/A280 ratio 1,81 +/- 0.05) within 40 min., while the mini spin colum kit (Nucleospin Quickpure) obtained lower yield but the best DNA quality (mean recovery 25.7 +/- 2.3 microg/ml; A (260)/ A (280) ratio = 1,83 +/- 0.06) with fewer processing time (25 min). Costs of each extraction varied from 3.28 Euro for Genomic to 3.6 Euro for Nucleospin. Microwave radiation and resin-based method (GeneFizz) were single step/single tube procedures, that provided template DNA suitable for ASA-PCR assay, without any purification steps. The costs varied from 0.12 Euro for microwave to 1,23 Euro for resin based procedure. In conclusion, our alternative procedures were much faster (<15 min per extraction) and convenient (5.00-7.00 Euro per test) but equally sensitive compared to conventional DNA extraction methods. Moreover, these procedures are easily adaptable to the routine processing of high number of clinical samples and coupled with ASA-PCR assay result particularly suitable for a large scale screening for the diagnosis and prevention of the thrombotic risk.

DNA↗

PubMed2003

Decreased hypocretin-1 (Orexin-A) levels in the cerebrospinal fluid of patients with myotonic dystrophy and excessive daytime sleepiness.

STUDY OBJECTIVE: Myotonic dystrophy type 1 is a multisystem disorder with myotonia, muscle weakness, cataracts, endocrine dysfunction, and intellectual impairment. This disorder is caused by a CTG triplet expansion in the 3' untranslated region of the DMPK gene on 19q13. Myotonic dystrophy type 1 is frequently associated with excessive daytime sleepiness, sharing with narcolepsy a short sleep latency and the presence of sleep-onset rapid eye movement periods during the Multiple Sleep Latency Test. Since narcolepsy is characterized by a dysfunction of the hypothalamic hypocretin system, we investigated whether patients with myotonic dystrophy type 1 with excessive daytime sleepiness have abnormalities in the hypocretin system. DESIGN/PARTICIPANTS: Six patients with myotonic dystrophy type 1 complaining of excessive daytime sleepiness and 13 healthy controls without a sleep disorder were included. The patients with myotonic dystrophy type 1 were evaluated using clinical interviews, nocturnal polysomnograms, and Multiple Sleep Latency Tests. All patients had a confirmed genetic diagnosis for DM1 and were HLA typed. Cerebrospinal fluid hypocretin-1 levels were measured using a direct radioimmunoassay in patients and controls. SETTING: University hospital sleep laboratory. INTERVENTIONS: N/A. MEASUREMENT AND RESULTS: The mean sleep latency on Multiple Sleep Latency Tests was abnormal in all patients (< 5 minutes in 2, < or = 8 in 4) and 2 sleep-onset rapid eye movement periods were observed in 2 subjects. All patients were HLA-DQB1*0602 negative. Hypocretin-1 levels were significantly lower in patients versus controls (p < 0.001); 1 case with 2 sleep-onset rapid eye movement periods had hypocretin-1 levels in the range generally observed in narcolepsy (< 110 pg/mL). Three cases had intermediate levels (110-200 pg/mL). Hypocretin-1 levels did not correlate clinically with disease severity or duration or with subjective or objective sleepiness reports. CONCLUSIONS: A dysfunction of the hypothalamic hypocretin system may mediate sleepiness and abnormal Multiple Sleep Latency Test results in patients with myotonic dystrophy type 1.

Adult↗

PubMed2006

Evaluation of FDA licensed HIV assays using plasma from Cameroonian blood donors.

Several diagnostic assays for the detection of HIV infection have been approved and licensed by the FDA for blood donor screening. However, the performance of these assays is unknown when testing genetically divergent blood specimens. To evaluate the performance of these assays with diverse HIV strains, we chose to study specimens collected from blood donors in Cameroon where genetic diversity and recombinant variants are prevalent. In this study, we tested 240 human plasma specimens collected from two blood centers in Cameroon. These samples were screened initially in Cameroon for antibody to HIV using a rapid assay. We also performed sequencing to determine subtype. Our evaluation has demonstrated that HIV infection in most HIV plasma samples could be detected by most of the US FDA licensed diagnostic assays. With the exception of a few specimens, HIV-1 p24 antigen was not detected in any of the samples. In addition, some nucleic acid tests (NAT) assays were not able to detect a few serologic reactive samples and all new variants including some CRF02_AG variants.

Blood Donors↗

PubMed1977

Primary deficiencies in humoral immunity.

Patients with primary disorders of B lymphocytes and immunoglobulins usually display increased susceptibility to bacterial infections but atopic, autoimmune, and malignant disorders are also more common in these patients. The spectrum of these disorders ranges from a virtual absence of B cells and immunoglobulins to selective deficiencies of immunoglobulin subclasses. The diagnosis is dependent upon the demonstration of the immunologic deficits by specialized laboratory procedures which include the quantitation of immunoglobulins, the formation of antibodies in vivo and in vitro and the demonstration of B cells in the tissues or the peripheral blood. There are five major points in the management of these patients: (1) the delineation of the immunologic defects by laboratory testing, (2) the use of parenterally injected human immunoglobulins, (3) the rapid identification of infecting organisms and the prompt institution of appropriate antibiotic therapy, (4) screening the family for immunodeficiency in those cases which are of genetic origin, and (5) genetic counseling.

Adult↗

PubMed1994

Polymerase chain reaction: basic concepts and clinical applications in dermatology.

The polymerase chain reaction (PCR) has been extensively used in basic science research, and the clinical potential of PCR is only now beginning to be realized. The PCR is based on the fundamental DNA replication process that occurs in every living cell. PCR is essentially an in vitro adaptation of the in vivo DNA copying process. Because PCR is so efficient at amplifying even picogram quantities of DNA, contamination with even trace amounts of nucleic acids can lead to the generation of unwanted DNA sequences and false-positive test results. Despite this, there has been rapid growth in the use of PCR in biomedical research and clinical diagnostics. PCR is the most sensitive test for herpes simplex virus, varicella-zoster virus, and human papillomavirus infections. Other diagnostic uses, including tests for genetic diseases, cancers, and other infectious diseases, are evolving.

False Positive Reactions↗

PubMed2026

Utilisation and Perceived Value of Genetic Counsellors Within US Haemophilia Treatment Centres.

INTRODUCTION: Rapid advancement of molecular genetics has transformed the diagnosis, treatment, and management of individuals with hereditary bleeding disorders. To provide effective, up-to-date genetic counselling, navigate the complexity of these conditions, and select appropriate molecular testing, genetics expertise is required. AIM: This study assessed the provision of genetic counselling services, involvement of genetic counsellors (GCs), and the perceived value of GCs within haemophilia treatment centres (HTCs) in the United States. METHODS: A survey was emailed to 396 HTC providers. Of these, 115 responses were received, representing 68 of 149 US HTCs (45.6% HTC participation rate). Responses were stratified by level of GC engagement. RESULTS: Although GCs have extensive training in genetics, genomics and counselling skills, nearly one-third of respondents (34.9%, n = 38) reported that a GC is not involved with the HTC nor are referrals made. Almost all GC-engaged respondents (98%, n = 22) and GC-referral respondents (95%, n = 20) agreed that 'GCs have a unique skill set that is highly valuable to an HTC clinic' compared to only 62% (n = 20) of non-GC-engaged respondents (p = 0.001). Additionally, respondents noted positive implications of integrating a GC within their HTCs, stating that GCs are 'ideal for optimal patient care'. CONCLUSION: These results highlight the value of a GC within an HTC. This signifies the need to reassess the role of GCs among HTCs to reduce inconsistencies in provision of genetic counselling and increase healthcare equity.

Humans↗

PubMedDate not supplied

Protistan diversity estimates based on 18S rDNA from seawater incubations in the Western North Atlantic.

Cloning/sequencing and fragment analysis of ribosomal RNA genes (rDNA) are becoming increasingly common methods for the identification of microbial taxa. Sequences of these genes provide many additional taxonomic characters for species that otherwise have few distinctive morphological features, or that require involved microscopy or laboratory culture and testing. These same approaches are now being applied with great success in ecological studies of natural communities of microorganisms. Extensive information on the composition of natural microbial assemblages is being amassed at a rapid pace through genetic analyses of environmental samples and comparison of the resulting genetic information with well-established (and rapidly growing) public databases. We examined microbial eukaryote diversity in a natural seawater sample from the coastal western North Atlantic Ocean using two molecular biological approaches: the cloning and sequencing of rRNA genes and by fragment analysis of these genes using the terminal restriction fragment length polymorphism (T-RFLP) method. A simple experiment was carried out to examine changes in the overall eukaryote (largely protistan) diversity and species composition (phylotype diversity) of a natural microbial assemblage when a seawater sample is placed in a container and incubated at ambient light and temperature for 72 h. Containment of the natural seawater sample resulted in relatively minor changes in the overall eukaryote diversity (species richness) obtained by either molecular method at three time points (time-zero, time-24 h, time-72 h). However, substantial changes in the dominance of particular eukaryote phylotypes took place between the three sampling times. Only 18% of the total number of phylotypes observed in the study were observed at all three time points, while 65% (108 of 165) phylotypes were observed only at a single time point (54 unique phylotypes initially, 37 more unique phylotypes at 24 h, and 17 more at 72 h). The results of this study indicate that a high diversity of protistan taxa existed in the original seawater sample at very low abundance, and thus were not observed in the initial characterization of community structure. Containment resulted in significant shifts in the dominance of these taxa, enabling the presence of previously unobserved phylotypes to be documented after 24 or 72 h of incubation.

Animals↗