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Background  Brucellosis is one of the most severe widespread zoonoses caused by the Gram-negative bacterium  Brucella species . The diagnosis and clinical assessment of human brucellosis are very important for the management of patients, while there is a lack of effective methods to detect  Brucellae . Classical culture of  Brucella  species is time consuming and often fails. A simple and sensitive assay is needed for diagnosis of  Brucella  infection and monitoring of treatment in man.    Methods  Blood samples and peripheral blood mononuclear cells (PBMCs) were collected from 154 patients hospitalized for brucellosis.  Brucella  antibodies were detected by Rose Bengal Plate Test (RBPT), Standard Tube Agglutination Test (SAT) and enzyme-linked immunosorbent assay (ELISA). Intracellular  Brucellae  were detected by blood culture and immunofluorescence staining (IFS).    Results  Among 154 brucellosis patients, 59.7% (92/154) were antibody reactive by RBPT, 81.8% (126/154) by SAT and 95.5% (147/154) by ELISA, respectively. Only 3.2% (5/154) of patient blood samples resulted in positive  Brucella  culture, while 68.8% (106/154) carried IFS detectable  Brucella  antigens in PBMCs. Gender ( P  = 0.01) but not age ( P  > 0.05) was a significant risk factor. The frequency of intracellular  Brucella  antigens was similar between patients receiving different treatment regimens ( P  > 0.05). However, a significant decrease of intracellular  Brucellae  was observed only in patients with acute brucellosis after the third course of treatment ( P  < 0.05), suggesting that current regimens to treat chronic brucellosis were not effective.    Conclusions  IFS appears a sensitive assay for detection of  Brucella  antigens in PBMCs and could be used for diagnosis and therapeutic monitoring of brucellosis in clinical practice.

Detection of Brucellae in peripheral blood mononuclear cells for monitoring therapeutic efficacy of brucellosis infection