Implementation in Cuba of a molecular assay for the detection of Brucella spp. in humans
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Abstract
Brucellosis is a zoonosis that may cause mild symptoms or severe manifestations. Molecular techniques are very sensitive and specific in the timely and early detection of Brucella spp. DNA, both at the beginning of the infection and in relapses and therapeutic failures. The aim of the present research was to implement a conventional PCR-bcsp that would allow the timely diagnosis of human brucellosis in Cuba, and thus contribute to avoid the occurrence of complications that can lead to disabling sequelae in the patient. A service and system research was carried out at the National Reference Laboratory for Leptospira and Brucella at IPK to evaluate the PCR-bcsp. The parameters of analytical sensitivity and specificity, as well as diagnostic sensitivity and specificity were determined. Conventional PCR was applied to 86 sera from suspected cases of human brucellosis that were negative to the IgM and IgG ELISAs for Brucella. PCR-bcsp presented a detection limit of 6.403 fg/µL. Analytical specificity was 100 %, while the diagnostic sensitivity and specificity showed values of 96.3 % and 100 %, respectively. After its application to sera from serologically negative probable cases of human brucellosis, 85 % positivity was reached. The present research strengthened the microbiological diagnosis of human brucellosis since a molecular tool guaranteeing the early diagnosis of the disease in Cuban environment has been implemented.
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