Serodiagnosis of human brucellosis by an indirect ELISA test using recombinant outer membrane protein 19 kDa (rOMP19) as an antigen.

BACKGROUND: Brucellosis remains one of the global health concerns that reemerges in recent years. Delayed or inaccurate diagnosis end to a long treatment duration and financial burden; therefore, finding a good antigen for detection of specific anti-Brucella antibodies is crucial. We intended to evaluate the serodiagnosis value of recombinant Brucella outer membrane protein 19 kDa (rOMP19) using indirect ELISA system compared with Rose Bengal test. RESULTS: The OMP19 sequence was successfully cloned into pET-28a and produced in E. coli cells (DE3). After extraction and purification of rOMP19, this protein was used for designing indirect ELISA to detect anti-Brucella antibodies in 73 human sera, including 6 brucellosis-positive and 67 brucellosis-negative samples. The accuracy of rOMP19 ELISA was evaluated by receiver operating characteristic (ROC) curve and then compared with Rose Bengal plate test and a commercial anti-IgG Brucella ELISA kit. In comparison with Rose Bengal plate test, the area under the ROC curve was 0.985 (95% CI, 0.96-1.00). From coordinates of the curve, the optimal cut-off value was selected at 0.147, in which the diagnostic sensitivity was 100%, and the specificity was 94%. At this cut-off point, 10 samples were diagnosed as positive (6 true positives and 4 false positives), while negative samples were all correctly diagnosed. The results of our designed rOMP19 ELISA was the same as data obtained from commercial ELISA kit, which applied LPS as an antigen. CONCLUSIONS: We concluded that OMP19 is an efficient antigen for the serodiagnosis of human brucellosis.

Molecular detection, genetic diversity, and phylogenetic analysis of foot-and-mouth disease virus (FMDV) type O in Iran during 2015-2016.

Background: The major challenge of foot-and-mouth disease (FMD) control is attributed to the rapid mutations in the FMDV RNA genome, resulting in continuous antigenic changes of circulating strains. Despite widespread vaccination of livestock populations, the incidence of the FMDV serotype O outbreaks in Iran during 2015-2016 raised concerns about the emergence of new strains. Aims: The aim of this study is the genetic and antigenic evaluation of FMDV type O isolates from different outbreak areas including Alborz, Tehran, Isfahan, Markazi, Zahedan, and Qom provinces. Methods: For this purpose, 71 FMD-infected samples were collected from six provinces of Iran, of which 12 serotype O positive were selected for genetic analysis. Results: All samples were in ME-SA topotypes/OPanAsia2 lineage, and the overall mean of genetic diversities at the 1D gene level was about 5% between the sequences. Blasting 1D gene sequences of isolated viruses showed more than 90% genetic identity with sequences registered from neighboring countries; therefore, it could be concluded that they had a common origin. Six isolates showed the highest genetic diversity (6% to 11%) with the OPanAsia2 vaccine strain (JN676146), which three of them (Qom, Alborz, and Zahedan isolates), had less than 30% antigenic homology with the OPanAsia2 virus (JN676146). Conclusion: Results of this study suggested OPanAsia2 vaccine had no enough coverage with some circulating strains in outbreak areas in Qom, Alborz, and Zahedan provinces, and the necessity of OPanAsia2 replacement with a new vaccine strain in Iran.

Rhodanese activity in different tissues of the ostrich.

1. The purpose of this investigation was to determine the activity, and compare the pattern of distribution, of rhodanese (thiosulphate: cyanide sulphurtransferase, EC. 2.8.1.1) in different tissues of male and female ostriches. 2. Tissue samples from male and female Blue Neck ostriches were assayed for rhodanese activity by the determination of thiocyanate formed by the action of the enzyme on thiosulphate and KCN. 3. Rhodanese was present in all tissues, and the highest activity was observed in the kidney and liver. Other tissues which contained significant activities of rhodanese were the duodenum, pancreas, heart, caecum and rectum. 4. Unlike other birds, the proventiculus does not appear to have an important role in cyanide detoxification in ostrich and, like mammals, the kidney and liver perform this function. 5. The results suggest that the main organs harbouring high rhodanese activity in the ostrich are associated with sites likely to be required in rhodanese mediated cyanide detoxification.