Diagnosis of recent and relapsed cases of human brucellosis by PCR assay

BACKGROUND: Brucellosis affects human populations in many developing countries including the Middle East, and Latin America where it is still endemic. It has been prevalent in Jordan for years, where 7842 cases of human brucellosis were registered at the Ministry of Health during 10 year-period. This study was initiated by the recent increase in the number of human cases diagnosed in a rural area in the Northern Jordan to help assess the status of the disease in that area. For this purpose blood specimens from brucellosis suspected cases were tested by serology, culture and PCR. METHODS: Peripheral blood specimens from 50 healthy control subjects and 165 seropositive patients having compatible signs and symptoms that were clinically diagnosed to have brucellosis were tested by blood culture, and by PCR. The PCR assay used genus-specific primers from the conserved region of the 16S rRNA sequence, which showed high specificity for the Brucella spp. RESULTS: Diagnosis of Brucella was established by PCR in 120 cases (72.7%). All of them were seropositive and 20 were positive by culture. Forty-eight of 58 (82.8%) of the relapsed cases two months after completing the treatment with an increase in the previous serological titers were positive by PCR. The assay has 85.7% positive predicative value, 100% sensitivity and specificity since it correctly identified all cases that were positive by blood cultures, 95.8% by serology and none of the control group was positive. CONCLUSIONS: Results showed that PCR assay can be applied with serology for the diagnosis of brucellosis suspected cases and relapses regardless of the duration or type of the disease without relying on the blood cultures, especially in chronic cases

Conditions influencing the in vitro antifungal activity of lactoferrin combined with antimycotics against clinical isolates of Candida - Impact on the development of buccal preparations of lactoferrin

Lactoferrin, an iron-binding glycoprotein, is a potential agent for the treatment of oropharyngeal Candidiasis. The aim of the present study was to test the capability of lactoferrin, combined or not combined with conventional antifungal agents, to inhibit the growth of different Candida species under various experimental conditions to be of guidance in the development of a suitable pharmaceutical formulation containing lactoferrin. The anti-Candida activities of lactoferrin were considerably higher using RPMI instead of SLM as assay medium. They were moreover increased by raising the medium pH from 5.6 to 7.5. With the 'standard' antifungal agent fluconazole similar results were found as for lactoferrin, but the medium type and pH did not affect MIC values of amphotericin B. The addition of saliva to medium did not reduce the antifungal activities of the individual compounds. Synergistic inhibitory effects on Candida growth were found for combinations of lactoferrin and fluconazole or amphotericin B, irrespective of the medium type and pH, or the addition of saliva. This indicates that for treatment of oral Candidiasis a formulation containing lactoferrin seems appropriate; results may be optimized if the formulation is provided with buffer capacity to attain pH 7.5 in the mucosal fluid. The synergistic effects between lactoferrin and 'standard' antifungals indicate that combinations should be considered in such a formulation