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histoplasmosis. In humans, whole blood RT-PCR has a reported sensitivity of 18% to 97% for the diagnosis of disseminated histoplasmosis, depending on whether a single or multiple blood samples are assayed per patient. 14 – 16 Although whole blood Histoplasma

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in Journal of the American Veterinary Medical Association

analysis, ACE, and RT-PCR assays. 6 More recently, ACE and RT-PCR assays have become widely used as screening tests because of their low cost, ease of use, adaptation for testing a high volume of samples, and analytic sensitivity. The RT-PCR assay has the

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in American Journal of Veterinary Research

transmission of A marginale during simulated vaccination between needle-free and conventional needle injection techniques and to evaluate the diagnostic efficacy of light microscopy, a cELISA, and an A marginale -specific RT-PCR assay. Materials and

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in American Journal of Veterinary Research

curves derived as a result of competitive RT-PCR, and each expression ratio was normalized by use of the PTEN gene expression level in nonneoplastic mammary gland samples (ratio, 1.0). Each expression ratio represented the mean of the results of 2

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in American Journal of Veterinary Research

relationship between genomes was determined on the basis of their proximity on the resulting phylogenetic tree. The tree was rooted at the phylogenetic midpoint. Assessment of FCV RT-PCR primers from previous studies To explore the effect that detection

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in American Journal of Veterinary Research

) specimens fixed in neutral-buffered 10% formalin, and an RT-PCR assay. Materials and Methods Cattle and sample collection —After weaning, calves weighing approximately 227 kg (499 lb) were processed at a southwest Kansas feedlot prior to placement in a

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in Journal of the American Veterinary Medical Association

succumbed to infection or were euthanized via pentobarbital overdose due to endpoint criteria (moribund, anorexic > 2 days, dyspneic, hemorrhagic discharge from nose or mouth) were necropsied and livers saved for by real-time reverse transcriptase (RT-PCR

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in American Journal of Veterinary Research

canine ASCs can differentiate into endothelial lineages. Therefore, the purpose of the study reported here was to evaluate whether canine ASCs differentiate into endothelial lineages, as determined by use of morphological, immunophenotypic, RT-PCR assay

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in American Journal of Veterinary Research

Abstract

Objective

To increase the timeliness and sensitivity of a procedure that uses viral nucleic acid amplification followed by restriction fragment length polymorphism (RFLP) analysis for identifying strains of porcine reproductive and respiratory syndrome virus (PRRSV).

Sample Population

24 strains of PRRSV.

Procedure

A nested-set reverse transcriptase-polymerase chain reaction (RT-PCR) was developed and compared with a nonnested-set RT-PCR for sensitivity in amplifying known quantities of infective PRRSV. Once reaction conditions were optimized, the nested-set RT-PCR was tested for effectiveness with 24 strains of PRRSV isolated from swine.

Results

The nested-set RT-PCR was 100- to 1,000- fold more sensitive than the nonnested-set RT-PCR, detecting as little as 1 infective unit of PRRSV/ml of sample. It also was generally as sensitive as the combination of steps, namely virus isolation or propagation and nonnested-set RT-PCR, currently used routinely for amplifying PRRSV prior to RFLP analysis, and it was effective for amplifying all of the 24 strains of PRRSV tested. Using this RT-PCR, all tests were completed within 1.5 days (including RFLP analysis), compared with the > 7 days often required for the currently used method involving virus isolation and propagation.

Conclusions

The nested-set RT-PCR was generally as sensitive as the combination of methods now used for PRRSV amplification prior to RFLP analysis, and it can markedly reduce the time required for testing.

Clinical Relevance

Presumptive identification of PRRSV strains can be provided in a more timely manner by use of a nested-set RT-PCR. (Am J Vet Res 1999;60:802–806)

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in American Journal of Veterinary Research

, Spinsanti G , Silvestrelli M , Supplizi AV . Exercise induced stress in horses: selection of the most stable reference genes for quantitative RT-PCR normalization . BMC Mol Biol . 2008 ; 9 : 49 . doi: 10.1186/1471-2199-9-49 18489742 28

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in Journal of the American Veterinary Medical Association