Visual detection of turkey coronavirus RNA in tissues and feces by reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with hydroxynaphthol blue dye

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development of a Loop-Mediated Isothermal Amplification Method for Detection of Histoplasma capsulatum DNA in Clinical Samples

Improved methods for the detection of Histoplasma capsulatum are needed in regions with limited resources in which the organism is endemic, where delayed diagnosis of progressive disseminated histoplasmosis (PDH) results in high mortality rates. We have investigated the use of a loop-mediated isothermal amplification (LAMP) assay to facilitate rapid inexpensive molecular diagnosis of this disease. Primers for LAMP were designed to amplify the Hcp100 locus of H. capsulatum. The sensitivity and limit of detection were evaluated using DNA extracted from 91 clinical isolates of known geographic subspecies, while the assay specificity was determined using DNA extracted from 50 other fungi and Mycobacterium tuberculosis. Urine specimens (n = 6) collected from HIV-positive individuals with culture- and antigen-proven histoplasmosis were evaluated using the LAMP assay. Specimens from healthy persons (n = 10) without evidence of histoplasmosis were used as assay controls. The Hcp100 LAMP assay was 100% sensitive and specific when tested with DNA extracted from culture isolates. The median limit of detection was <= 6 genomes (range, 1 to 300 genomes) for all except one geographic subspecies. The LAMP assay detected Hcp100 in 67% of antigen-positive urine specimens (4/6 specimens), and results were negative for Hcp100 in all healthy control urine specimens. We have shown that the Hcp100 LAMP assay is a rapid affordable assay that can be used to expedite culture confirmation of H. capsulatum in regions in which PDH is endemic. Further, our results indicate proof of the concept that the assay can be used to detect Histoplasma DNA in urine. Further evaluation of this assay using body fluid samples from a larger patient population is warranted.

Detection of Trypanosoma vivax using PCR and LAMP during aparasitemic periods

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation and characterization of ten novel microsatellite loci in the red-winged tinamou (Rhynchotus rufescens, Tinamiformes, Aves) and cross-amplification in other tinamous

We describe the isolation and characterization of ten microsatellite loci from the red-winged tinamou (Rhynchotus rufescens) and also evaluated the cross-amplification of these loci and other ten loci previously developed for the great tinamou (Tinamus major) in other tinamous. Genetic variability was assessed using 24 individuals. Six loci were polymorphic with moderate to high number of alleles per locus (2-12 alleles) and showed expected heterozygosity (HE) ranging from 0.267 to 0.860. All loci conformed to the Hardy-Weinberg expectation and linkage disequilibrium was not significant for any pair of loci. This battery of polymorphic loci showed high paternity exclusion probability (0.986) and low genetic identity probability (4.95 x 10(-5)), proving to be helpful for parentage tests and population analyses in the red-winged tinamou. The cross-amplification was moderate where of the 160 locus/taxon combinations, 46 (28.75%) successfully amplified.

Strain differentiation of Trichophyton rubrum by random amplification of polymorphic DNA (RAPD)

Trichophyton rubrum é um importante agente causal de dermatomicose. Os métodos de tipagem molecular têm sido recentemente desenvolvidos para responder questões sobre epidemiologia e auxiliar no esclarecimento de recidivas, após o tratamento. As seqüências aleatórias 1- (5'-d[GGTGCGGGAA]-3') e 6- (5'-d[CCCGTCAGCA]-3') foram usadas para tipagem molecular deste fungo por RAPD produzindo variabilidade intraespecífica. Cinco padrões foram observados entre os 10 isolados de T. rubrum, com ambas as seqüências. Foi concluído que a análise por RAPD pode ser utilizada para estudos epidemiológicos.

A simple interferometric method to measure the calibration factor and displacement amplification in piezoelectric flextensional actuators

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Strategic capacitor placement in distribution systems by minimisation of harmonics amplification because of resonance

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A comparative study of the non-isothermal degradation of natural rubber from Mangabeira (Hancornia speciosa Gomes) and Seringueira (Hevea brasiliensis)

Due to the increasing search for alternative sources of natural rubber (NR) whose properties are similar to Hevea brasiliesis, several sources have been studied in the past few years. Among them, Mangabeira (Hancornia speciosa Gomes), which is native to Amazon rainforest and other regions of Brazil, has a potential as another viable rubber source. As a continuation of a series of comparative studies between Hancornia and Hevea (clone RRIM 600) these two species by our research team, their thermal behavior was analyzed by thermogravimetry (TG) using Flynn-Wall-Ozawa's approach in order to obtain kinetic parameters (reaction order, pre-exponential factor and activation energy) of the decomposition process. Results indicated that the thermal behavior of NR from Hancornia was comparable to Hevea with some differences observed as follows: reaction order for Hancornia was higher than for Hevea at the beginning of degradation and very close for temperatures over 350 A degrees C; activation energy and pre-exponential factor had the same trend, i.e., increased with increasing degree of conversion remaining almost constant between 20 and 70% and then increasing for higher degrees, although Hevea was slightly more thermally stable than Hancornia. These major influences in the degradation process in the early stage are attributed to differences in non-rubber constituents present in these two species.

NON-ISOTHERMAL STUDIES on CRYSTALLIZATION KINETICS of TELLURITE 20Li(2)O-80TeO(2) GLASS

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A set of polymorphic microsatellite loci for Vriesea gigantea and Alcantarea imperialis (Bromeliaceae) and cross-amplification in other bromeliad species

Fifteen polymorphic microsatellite markers were isolated and characterized in two species of Bromeliaceae: Vriesea gigantea and Alcantarea imperialis. The number of alleles observed for each locus ranged from three to 16. The loci will be used for studies of the genetic structure of natural populations, reproductive biology, and evolutionary relationships among and within these genera. A cross-amplification test in 22 taxa suggests that the markers will be useful for similar applications in numerous other bromeliad species.

Isolation and characterization of eight microsatellite loci in Leporinus macrocephalus (Characiformes : Anostomidae) and cross-species amplification

Leporinus macrocephalus is a species endemic to the Paraguay River basin and an important fishery resource, as well as a valuable species in aquaculture programs. A total of eight polymorphic microsatellite loci were isolated and characterized. A population survey was conducted involving 45 specimens whereby a large number of alleles (range 5-17 among loci), a highly observed (0.1667-0.6129) and an expected (0.6967-0.9448) heterozygosity was detected, indicating its usefulness in population genetics studies. Cross-species amplification was successful in eight Characiformes species.

Isolation and characterization of microsatellite loci in Pseudoplatystoma corruscans (Siluriformes : Pimelodidae) and cross-species amplification

A total of five polymorphic microsatellites loci from Pseudoplatystoma corruscans were isolated and characterized. A population survey involving 43 specimens resolved a large number of alleles (range seven to eight among loci) and high observed heterozygosity (0.500-0.615), indicating its usefulness in population genetics studies. Cross-species amplification was successful in four other Pimelodidae species.

Identification and characterization of polymorphic microsatellite loci in the blue shark Prionace glauca, and cross-amplification in other shark species

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development and characterization of microsatellite loci in Phractocephalus hemioliopterus (Siluriformes: Pimelodidae) and their cross-species amplification in six related species

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Isolation and characterization of microsatellite loci in the Neotropical fish Astyanax altiparanae (Teleostei: Characiformes) and cross-species amplification

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)