RESUMO
Members of the genus Brucella are known worldwide as pathogens of wildlife and livestock and are the most common organisms of zoonotic infection in humans. In general, brucellae exhibit a range of host specificity in animals that has led to the identification of at least seven Brucella species. The genomes of the various Brucella species are highly conserved, which makes the differentiation of species highly challenging. However, we found single-nucleotide polymorphisms (SNPs) in housekeeping and other genes that differentiated the seven main Brucella species or clades and thus enabled us to develop real-time PCR assays based around these SNPs. Screening of a diverse panel of 338 diverse isolates with these assays correctly identified each isolate with its previously determined Brucella clade. Six of the seven clade-specific assays detected DNA concentrations of less than 10 fg, indicating a high level of sensitivity. This SNP-based approach places samples into a phylogenetic framework, allowing reliable comparisons to be made among the lineages of clonal bacteria and providing a solid basis for genotyping. These PCR assays provide a rapid and highly sensitive method of differentiating the major Brucella groups that will be valuable for clinical and forensic applications.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Brucella/classificação , Brucella/genética , DNA Bacteriano/genética , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Nucleotídeo Único , Animais , DNA Bacteriano/química , Genótipo , Humanos , Dados de Sequência Molecular , Sensibilidade e Especificidade , Análise de Sequência de DNARESUMO
Allergic rhinitis is a common health problem usually treated with drug therapy. Some patients experience side effects of drug therapy while others fear the use of drugs. Acupuncture is an interesting alternative to traditional treatment. The few studies evaluating acupuncture indicate a possible clinical effect on allergic rhinitis. This study compared active versus sham acupuncture in 40 consecutive patients with a history of allergic rhinitis and a positive skin test. Patients were randomized and assessed prior to treatment and then reassessed after 12 months. Improvements in symptoms using visual analogue scales, reduction in skin test reactions and levels of specific immunoglobin E (IgE) were used to compare the effect of treatment. For one allergen, mugwort, a greater reduction in levels of specific IgE (p = 0.019, 0.039) and skin test reaction (p = 0.004) was seen in the group receiving active acupuncture compared to the group receiving sham acupuncture. However, this finding might be an artifact. No differences in clinical symptoms were seen between active versus sham acupuncture, thus the conclusion being that the effect of acupuncture on allergic rhinitis should be further evaluated in larger randomized studies.