Search results for "Nucleic acid amplification technique"
showing 10 items of 26 documents
Fast detection of Southern tomato virus by one-step transcription loop-mediated isothermal amplification (RT-LAMP)
2017
Southern tomato virus (STV) is a double stranded RNA (dsRNA) virus belonging to genus Amalgavirus (family Amalgamaviridae) which has been detected in tomato plants showing stunting, fruit discoloration and size reduction. A one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of STV in total RNA or sap extracts (obtained just by grinding in buffer) from STV-infected tomato plants by using a set of three primers pairs which were designed to the sequence of the STV putative coat protein. Amplification products were visualized by gel electrophoresis or direct staining of DNA. The sensitivity of RT-LAMP was identical to that of th…
Performance comparison of two whole genome amplification techniques in frame of multifactor preimplantation genetic testing
2018
Purpose To compare multiple displacement amplification and OmniPlex whole genome amplification technique performance during array comparative genome hybridization (aCGH), Sanger sequencing, SNaPshot and fragment size analysis downstream applications in frame of multifactor embryo preimplantation genetic testing. Methods Preclinical workup included linked short tandem repeat (STR) marker selection and primer design for loci of interest. It was followed by a family haplotyping, after which an in vitro fertilization preimplantation genetic testing (IVF-PGT) cycle was carried out. A total of 62 embryos were retrieved from nine couples with a confirmed single gene disorder being transmitted in t…
Virología: diagnóstico sindrómico de meningitis y encefalitis
2020
Viral infections of the nervous system represent a major health problem. It is estimated that the incidence of viral meningitis in the general population ranges from 5-17 cases per 100,000 inhabitants per year in developed countries. This heading encompasses highly varied clinical pictures, ranging from meningitis to encephalitis. This article presents the agents involved in our environment and discusses their expressiveness. In immunocompetent patients, the course of these infections is usually benign. Nucleic acid amplification techniques are the gold standard for their etiological diagnosis. The introduction of polymerase chain reaction and serological diagnosis of the main arboviruses h…
Epigenetic differences arise during the lifetime of monozygotic twins.
2005
Monozygous twins share a common genotype. However, most monozygotic twin pairs are not identical; several types of phenotypic discordance may be observed, such as differences in susceptibilities to disease and a wide range of anthropomorphic features. There are several possible explanations for these observations, but one is the existence of epigenetic differences. To address this issue, we examined the global and locus-specific differences in DNA methylation and histone acetylation of a large cohort of monozygotic twins. We found that, although twins are epigenetically indistinguishable during the early years of life, older monozygous twins exhibited remarkable differences in their overall…
Evaluation of saliva as a complementary technique to the diagnosis of COVID-19:a systematic review
2021
Background Infectious disease coronavirus 2019 (COVID-19) is caused by the SARS-CoV-2 virus, and it mainly affects the upper respiratory tract. The gold standard for its diagnosis is real-time reverse transcription polymerase chain reaction (RT-qPCR) performed on a nasopharyngeal swab. In contrast, testing saliva has significant advantages as a diagnostic method. Material and Methods We searched for articles evaluating saliva as a diagnostic method for COVID-19 on the PUBMED/MEDLINE, WEB OF SCIENCE, COCHRANE, and SCIELO platforms. We initially found 233 articles and 20 were selected for inclusion following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses protocol: 18 c…
Role of one-step nucleic acid amplification (OSNA) to detect sentinel lymph node low-volume metastasis in early-stage cervical cancer.
2019
IntroductionGrowing evidence in the literature supports the accuracy of sentinel lymph node (SLN) biopsy in early-stage cervical cancer. One-step nucleic acid amplification (OSNA) is a rapid assay able to detect cytokeratin 19-mRNA in SLNs, and it can be used for intra-operative detection of low-volume metastases. The aim of this study was to evaluate the rate of low-volume metastasis in SLNs detected by OSNA in patients with early-stage cervical cancer. Secondary aims were to define the sensitivity and the negative predictive value of SLN biopsy assessed with OSNA.MethodsAfter IRB approval, consecutive patients who underwent surgery for International Federation of Gynecology and Obstetrics…
Growth of immobilized DNA by polymerase: bridging nanoelectrodes with individual dsDNA molecules.
2011
We present a method for controlled connection of gold electrodes with dsDNA molecules (locally on a chip) by utilizing polymerase to elongate single-stranded DNA primers attached to the electrodes. Thiol-modified oligonucleotides are directed and immobilized to nanoscale electrodes by means of dielectrophoretic trapping, and extended in a procedure mimicking PCR, finally forming a complete dsDNA molecule bridging the gap between the electrodes. The technique opens up opportunities for building from the bottom-up, for detection and sensing applications, and also for molecular electronics.
DNA Amplification Fingerprinting for Subtyping Neisseria gonorrhoeae Strains
1995
Background and Objectives DNA amplification fingerprinting is used in most epidemiologic studies as a substitute for conventional typing methods. DNA amplification fingerprinting and conventional typing methods were compared in this epidemiologic study of Neisseria gonorrhoeae. Goal of This Study To differentiate 70 Neisseria gonorrhoeae isolates from untreated patients with urogenital gonococcal infection. Study Design Gonococcal strains were characterized by auxo-typing, serotyping, plasmid profile, antibiotic sensitivity, and DNA amplification fingerprinting. The method of unweighted pair-group average linkage was used for cluster analysis. Discriminatory power was calculated applying Si…
Use of nodulation pattern, stress tolerance, nodC gene amplification, RAPD-PCR and RFLP-16S rDNA analysis to discriminate genotypes of Rhizobium legu…
2005
Twenty-seven new Rhizobium isolates were obtained from root nodules of wild and crop legumes belonging to the genera Vicia, Lathyrus and Pisum from different agroecological areas in central and southern Italy. A polyphasic approach including phenotypic and genotypic techniques was used to study their diversity and their relationships with other biovars and species of rhizobia. Analysis of symbiotic properties and stress tolerance tests revealed that wild isolates, showed a wide spectrum of nodulation and a marked variation in stress tolerance compared with reference strains tested in this study. All rhizobial isolates (except for the isolate CG4 from Galega officinalis) were presumptively i…
Identification and typing of food-borne Staphylococcus aureus by PCR-based techniques.
2005
Abstract The possibility of using PCR for rapid identification of food-borne Staphylococcus aureus isolates was evaluated as an alternative to the API-Staph system. A total of 158 strains, 15 S. aureus , 12 other staphylococcal species, and 131 isolates recovered from 164 food samples were studied. They were phenotypically characterized by API-Staph profiles and tested for PCR amplification with specific primers directed to thermonuclease ( nuc ) and enterotoxin ( sea to see ) genes. Disagreement between the PCR results and API-Staph identification was further assessed by the analysis of randomly amplified polymorphic DNA (RAPD) profiles obtained with three universal primers (M13, T3, and T…