Navegando por Palavras-chave "reação em cadeia da polimerase em tempo real"

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    Polimorfismos dos genes nr3c1, chrm2, tph1 e mc1r e sua correlação com marcadores de ancestralidade e fatores biopsicossociais em indivíduos com disfunção temporomandibular
    (Universidade Federal de São Paulo (UNIFESP), 2015-05-28) Ferreira, Mariana Brandao [UNIFESP]; Alonso, Luis Garcia Alonso [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Objectives: To assess a sample of 105 patients with temporomandibular joint dysfunction (TMD) and 210 controls (control group) for the following Research Diagnostic Criteria (RDC) Axis II biopsychosocial factors: depression, pain grades, and unspecific physical symptoms with and without pain, and to determine whether certain genetic polymorphisms increase susceptibility to these factors. Methods: RDC Axis I was used for diagnosing myofascial pain in the cases. Cases and controls answered the RDC Axis II questionnaire. The DNA of cases and controls was collected and quantified. Ancestry-informative markers were identified followed by polymorphism analysis of the genes NR3C1, CHRM2, TPH1, and MC1R using TaqMan Low Density Array (TLDA) personalized cards. Results: Polymorphisms of the genes NR3C1, CHRM2, TPH1, and MC1R were not associated with depression, pain, unspecific physical symptoms with and without pain, ancestry-informative markers, or TMD. Conclusions: The multifactorial polygenic mechanism of susceptibility is based on the idea that each gene contributes with small, additive, and relatively equal effects. Hence, it is possible that the study genes have more effective interrelations with other predisposing and regulatory genes, but in isolation they do not have enough biological and statistical consistency to explain TMD. By extension, the small number of Blacks, Natives, and Asians in this sample also corroborate to the weak association between these ethnicities factors and genetic polymorphisms. Larger studies, such as the ones currently being conducted by our group, can elucidate the associations postulated above more clearly.
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    O uso da reação em cadeia da polimerase em tempo real no diagnóstico de uveítes infecciosas
    (Universidade Federal de São Paulo (UNIFESP), 2014-09-30) Santos, Fabio Felipe dos [UNIFESP]; Mattos Junior, Rubens Belfort [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Infectious uveitis is a disease of major morbidity and blindness in several countries. Real-time PCR (qPCR) has been demonstrated as a high sensitivity diagnostic method of infections caused by parasites, viruses, fungi and bacteria. The technique allows the diagnosis of atypical infectious uveitis that cannot be diagnosed and differentiated only by clinical examination. The objective of this study was to analyze the qPCR technique in the detection of Herpes simplex virus (HSV), varicella zoster (VZV), cytomegalovirus (CMV), Toxoplasma gondii (TOXO), and M. tuberculosis (TB), for the differential diagnosis of patients with infectious uveitis. This study was divided into two experiments: In the first experiment, 27 patients with posterior infectious uveitis were recruited. In the second, 90 patients were recruited, of which 65 were diagnosed with ocular toxoplasmosis, and 25 patients were used as controls (infectious uveitis from other causes and not infectious). All patients were from the Department of Ophthalmology and Visual Sciences of the EPM/UNIFESP,São Paulo-Brazil. DNA extraction in the blood, aqueous humor and vitreous humor were performed. Experiment I showed T. gondii, CMV, VZV or HSV present in 19.2% of the aqueous humor samples and in 30% of the vitreous samples. Only CMV was detected in the blood (3.7%). The qPCR was able to confirm the diagnosis in 26% of patients with infectious posterior uveitis. In experiment II, from the total of 65 patients, 57% were positive for T. gondii, considering all the analyzed samples (blood, aqueous or vitreous). The vitreous and the aqueous were positive in respectively 40% and 58% of the samples and the blood test was positive in 25%. In the group of toxoplasmosis where blood and aqueous were collected, qPCR was positive in 68% of patients: 45.1% in the aqueous only, and 19.4% were positive in blood and aqueous simultaneously. In the Toxoplasmosis group with samples from vitreous, aqueous and blood were studies, the qPCR was positive in 41.7%: 16.7% only in the vitreous, 16.7% in the aqueous and vitreous, and 8.3% in the blood and aqueous. In the first experiment qPCR was able to detect the DNA of pathogens with higher sensitivity in the vitreous humor. In the second experiment, the sensitivity was higher in the aqueous humor. The qPCR in blood samples allowed to detect the DNA of pathogens causing infectious uveitis, however, due to low sensitivity, was not a useful sample to complement the clinical diagnosis. In conclusion, qPCR can be a useful tool for the diagnosis of infectious uveitis, assisting in proper treatment of ocular diseases. This study showed that the ocular fluids have better accuracy in molecular diagnosis of infectious uveitis, and the aqueous humor shown with positive differential for being less invasive during collection.