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Ellipsometric-based novel DNA biosensor for label-free, real-time detection of Bordetella parapertussis.

Pertussis (or whooping cough) is a contagious disease mainly affecting infants and children and predominantly caused by Bordetella pertussis followed by Bordetella parapertussis. B. parapertussis causes a milder cough but usually symptomatically appears like B. pertussis infection. Thus the epidemiology of illness caused by B. parapertussis is not well understood. In this study, a sensitive and specific method for the rapid diagnosis of B. parapertussis is presented. The covalent immobilization of thiol-terminated DNA oligonucleotides (ss DNA SAM) on a silicon surface by disulfide bond formati

A many probes-one spot hybridization oligonucleotide microarray.

A variant of the hybridization oligonucleotide microarray, utilizing the principle of many probes-one spot (MPOS-microarrays), is proposed. A case study based on Orthopoxviruses (Variola, Monkeypox, and Ectromelia viruses) demonstrates a considerable increase in the fluorescence signal (up to 100-fold) when several oligonucleotide probes are printed to one spot. Moreover, the specificity of detection also increases (almost 1000-fold), allowing the use of probes that individually lack such high specificity. The optimal probes have a Tm of 32-37 °C and length of 13-15 bases. We suggest that the

Comparison of four methods using throat swabs to confirm rubella virus infection.

Laboratory tests are essential for confirming sporadic cases and outbreaks of rubella. Detection of rubella virus is often necessary to confirm rubella cases and to identify specimens to be used to characterize wild-type rubella viruses. The sensitivities of four methods for detecting rubella virus infection using throat swabs, which had been collected in Henan and Anhui provinces in China, were evaluated. The methods used were reverse transcription (RT)-PCR followed by Southern hybridization using RNA extracted directly from clinical specimens, virus growth in tissue culture followed by virus

Taxonomic evidence that serovar 7 of Erysipelothrix strains isolated from dogs with endocarditis are Erysipelothrix tonsillarum.

The levels of relatedness among strains of Erysipelothrix serovar 7 isolated from dogs with endocarditis were estimated by performing DNA-DNA hybridization experiments with the type strains of Erysipelothrix rhusiopathiae and Erysipelothrix tonsillarum. All the canine strains exhibited more than 81% hybridization with the type strain of E. tonsillarum but less than 13% hybridization with the type strain of E. rhusiopathiae. Based on DNA-DNA hybridization results we confirmed that serovar 7 of the isolates from dogs with endocarditis were conclusively identified as E. tonsillarum. These results

A species-specific repetitive sequence in Mycobacterium leprae DNA.

A 2.2-kilobase Mycobacterium leprae DNA insert fragment from a recombinant genomic library (pYA1065) was found to hybridize to at least 19 fragments of chromosomal M. leprae DNA by Southern hybridizations. The probe hybridized to identical fragments of chromosomal DNA from four M. leprae isolates (two from patients with leprosy, one from a naturally infected armadillo, and one from a naturally infected Mangabey monkey) whether the chromosomal DNA was digested with BamHI, BstEII, PstI, or SacI. The pYA1065 probe is specific for M. leprae; it did not hybridize to chromosomal DNA from 14 cultivab

Extracellular Vesicle-Lipid Hybrid Systems for RNA Delivery in Cancer: Structural Classification, Functional Delivery, and Translational Challenges.

RNA therapeutics offer considerable potential for cancer treatment. Their therapeutic application, however, remains limited by rapid degradation, inefficient cellular uptake, and restricted intracellular release. Extracellular vesicles (EVs) are cell-derived membrane vesicles that have been exploited as promising vehicles for drug delivery due to their high biocompatibility and low immunogenicity, whereas liposomes and lipid nanoparticles provide tunable lipid composition and efficient loading of exogenous nucleic acids. Combining these carriers has led to EV-lipid hybrid systems designed to i

Rapid diagnosis of periodontal infections: findings in AIDS patients.

A small number of bacterial pathogens in the human oral cavity cause the different forms of periodontal disease. Of the approximately two hundred different oral bacterial species, about a dozen have been associated with these diseases including localized juvenile periodontitis, rapidly progressing periodontitis, and adult periodontitis. These species include Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, Porphyromonas gingivalis, and Prevotella intermedia. Several rapid methods have been developed to detect these species in clinical samples. These include im

Enzymatic RNA amplification of the enteroviruses.

Enteroviruses are among the most common causes of childhood infection. Current diagnostic techniques are often too slow and too insensitive to benefit the patient optimally. This report describes a modified polymerase chain reaction technique by which enteroviral RNA can be amplified, over a few hours, to a level detectable by agarose mini-gel electrophoresis or nucleic acid hybridization or both. Three oligomeric regions of great homology among the enteroviruses were identified and designated as a potential primer pair and probe. With this combination, all 11 of the enterovirus serotypes test…

Isolation and characterization of cloned human DNA fragments carrying reiterated sequences common to both autosomes and the X chromosome.

Several recombinants were identified and purified from a cloned library of human DNA by virtue of their homology to DNA from a mouse-human hybrid cell line containing a single human chromosome, the X, and their lack of homology to mouse DNA. Three recombinants were characterized in detail, and all were homologous to reiterated DNA from the human X chromosome. These recombinants also were homologous to reiterated sequences on one or more human autosomes and, therefore, were not X chromosome specific. The recombinant DNA fragments homologous to human reiterated X DNA were the same fragments homo

Annual growth cycle observation, hybridization and forcing culture for improving the ornamental application of Paeonia lactiflora Pall. in the low-latitude regions.

Expanding the southern range of herbaceous peony (Paeonia lactiflora Pall.) is a meaningful and worthwhile horticultural endeavor in the Northern Hemisphere. However, high temperatures in winter seriously hinder the bud dormancy release and flowering of peony in the more southern areas of subtropical and tropical regions. Resource introduction and hybridization can contribute to creating new cultivars with high adaptability in a warmer winter climate. In this study, three representative cultivars of P. lactiflora were screened for flowering capabilities and their annual growth cycles were obse

Potential antifolate resistance determinants and genotypic variation in the bifunctional dihydrofolate reductase-thymidylate synthase gene from human and bovine isolates of Cryptosporidium parvum.

We have determined the nucleic acid sequences of a gene encoding the bifunctional enzyme dihydrofolate reductase-thymidylate synthase (DHFR-TS) from bovine and human AIDS isolates of Cryptosporidium parvum. THe DHFR-TS gene was isolated from genomic DNA libraries by hybridization with a probe amplified from C. parvum genomic DNA using generic TS primers in the polymerase chain reaction. Genomic Southern and electrophoretic karyotype analyses reveal C. parvum DHFR-TS is a single-copy gene on a 1200-kb chromosome. The DHFR-TS nucleic acid sequence contains no introns and the single 1563-bp open

Nucleosides and nucleotides. Part 214: thermal stability of triplexes containing 4'alpha-C-aminoalkyl-2'-deoxynucleosides.

In order to develop novel antigene molecules forming thermally stable triplexes with target DNAs and having nuclease resistance properties, we synthesized oligodeoxynucleotides (ODNs) with various lengths of aminoalkyl-linkers at the 4'alpha position of thymidine and the aminoethyl-linker at the 4'alpha position of 2'-deoxy-5-methylcytidine. Thermal stability of triplexes between these ODNs and a DNA duplex was studied by thermal denaturation. The ODNs containing the nucleoside 2 with the aminoethyl-linker or the nucleoside 3 with the aminopropyl-linker thermally stabilized the triplexes, wher

A Fluorescence in Situ Hybridization (FISH) Test for Diagnosing Babesiosis.

Apicomplexan parasites of the genus Babesia cause babesiosis in humans and animals. The microscopic examination of stained blood smears, detection of serum antibodies by immunoassays, and PCR-based identification of parasite nucleic acid in blood are common laboratory methods for diagnosing babesiosis. The present study evaluated a commercially available Babesia genus-specific fluorescence in situ hybridization (FISH) test for detecting Babesia parasites in blood smears. The FISH test detected Babesia duncani and Babesia microti, two common species that cause human infections in the USA, and o

A modified system for analyzing ionizing radiation-induced chromosome abnormalities.

The analysis of dicentric chromosomes in human peripheral blood lymphocytes (PBLs) by Giemsa staining is the most established method for biological dosimetry. However, this method requires a well-trained person because of the difficulty in detecting aberrations rapidly and accurately. Here, we applied a fluorescence in situ hybridization (FISH) technique, using telomere and centromere peptide nucleic acid (PNA) probes, to solve the problem of biological dosimetry in radiation emergency medicine. A comparison by a well-trained observer found that FISH analysis of PBLs for the dose estimation wa

[Detection using molecular biology techniques of Mycoplasma hominis and Ureaplasma urealyticum in urogenital samples].

Mycoplasma hominis and Ureaplasma urealyticum are species closely related to urogenital diseases such as pyelonephritis, nongonococcal urethritis, urinary calculi, epididymitis, pelvic inflammation, infertility, abortions and post-delivery fever. They can also cause pneumonia and meningitis in newborn infants. In this paper we used nucleic acid hybridization and polymerase chain reaction to analyze 22 samples from patients with different urogenital symptoms in order to detect mycoplasmas and ureaplasmas. We obtained 10 positive samples and 12 were negative. From positive samples we identified …

Молекулярный подход к интеровиральному диагнозу идиопатической кардиомиопатии и миокардита.

В некоторых случаях миокардит человека и расширенная кардиомиопатия считаются этиологическими агентами. Мюрин-модели острого коксакивируса B3 миокардита включают вирусы коксаки B в качестве возможных причин миокардита человека. Косвенные данные, свидетельствующие о том, что энтеровирусы в качестве причинно-следственных агентов являются сердечными заболеваниями человека, получены в результате сероологических исследований. В последнее время в результате анализа гибридизации нуклеиновой кислоты были получены прямые доказательства присутствия в организме сердечных тканей больного человека. Хотя да

Совпадение в аннотации оригинала
Enteroviruses are thought to be etiologic agents in some cases of human myocarditis and dilated cardiomyopathy. Murine models of acute coxsa … ntly, direct evidence for enteroviral presence in diseased human heart tissues has been obtained by nucleic acid hybridization analyses. Although the data suggest that enteroviral infections may be associated with 18% to 50% of cases of myocarditis or dilated cardiomyopathy, or both, causality has not been established. Unanswered questions remain regarding the specific iden …

Demonstration of Epstein-Barr virus in malignant non-Hodgkin's lymphomas.

Lymph nodes or tumor biopsies of 60 persons suspected of having a malignant lymphoma were examined for the presence of Epstein-Barr virus nuclear antigen (EBNA) by anticomplement immunofluorescence. In 8 cases the tissue specimens were also assayed for Epstein-Barr virus (EBV) DNA by nucleic acid hybridization. Serum samples of patients and controls were tested for EBV-related antibodies. The histological tests in 37 cases showed a malignant non-Hodgkin lymphoma, and in 23 cases a reactive lymphadenopathy. A Burkitt lymphoma of a European boy and a polymorphic centroblastoma contained EBNA and…

R-loop-derived cytoplasmic RNA-DNA hybrids activate an immune response.

R-loops are RNA-DNA-hybrid-containing nucleic acids with important cellular roles. Deregulation of R-loop dynamics can lead to DNA damage and genome instability1, which has been linked to the action of endonucleases such as XPG2-4. However, the mechanisms and cellular consequences of such processing have remained unclear. Here we identify a new population of RNA-DNA hybrids in the cytoplasm that are R-loop-processing products. When nuclear R-loops were perturbed by depleting the RNA-DNA helicase senataxin (SETX) or the breast cancer gene BRCA1 (refs. 5-7), we observed XPG- and XPF-dependent cy

Quantitative detection of oncogenic HPV DNA using hybrid capture to triage borderline and mildly dyskaryotic Papanicolaou smears.

OBJECTIVE: To investigate the use of hybrid capture to triage borderline and mildly abnormal PAP smears to either colposcopy or continued cytological surveillance. METHOD: A prospective cross-sectional study of 207 women with either mild or borderline dyskaryosis referred to the colposcopy clinic at the Whittington Hospital, London was done. Quantitative detection of oncogenic human papillomavirus deoxyribose nucleic acid was performed using the solution hybridization technique Hybrid Capture I (Digene Diagnostics). Positive or negative hybrid capture results were compared to colposcopically

[Virus and gastrointestinal infections].

Numerous viruses found in the gut are not associated with primary infection or disease of the gastrointestinal tract. Other groups or viruse … astrovirus detection in faecal samples include immunoassays such as Elisa and latex agglutination. Nucleic acid hybridization techniques have generally not proven to be substantially sensitive and the more sensitive techniques recently developed use the polymerase chain reaction (adenovirus) or the reverse transcription/polymerase chain reaction (astrovirus, calicivirus, c …