Objective To compare the phenol water method and the iTron kit in extracting lipopolysaccharides(LPS) from Brucella. Methods LPS were extracted from B. melitensis strain 16M using phenol water method and kit method. The purity and structure of LPS was compared by SDS-gel electrophoresis and silver stain. The activity of the extracted LPS was detected by limulus amebocyte lysate agglutination test, and the characteristics of the two methods were compared. Results LPS extracted from Brucella by both methods had a high purity. There was no significant difference in activity of LPS extracted by phenol water method and the iTron kit ( t'=1.270, P=0.332), which was (4.926±0.051) and (5.015±0.037) EU/ng, respectively. LPS extracted by phenol water method might lose some LPS at 17×10 ³ and 26×10 ³, while using the iTron kit we can get intact LPS in a convenient and safe way. Conclusion The iTron kit has advantages in extracting LPS from Brucella compared with the phenol water method.

Objective Three suspected brucellosis cases were investigated in Suizhou city of Hubei province, and three suspected Brucella strains were identified. Methods Field investigation was applied to finish Brucella epidemiological survey, used Brucella conventional appraisal classification method and the phage cracking test for testing. BCSP31-PCR method (Based on Brucella genus specific gene BCSP31 target gene detection method), AMOS-PCR (based on electrophoresis banding identify B. abortus biovar 1, 2, 4 of Brucella, B. melitensis, B. ovis and B. suis biovar 1) and Real-time PCR, three molecular classification method were compared. Results Three patients were infected by close contact with sick animals and their meat, conventional identification and a variety of polymerase chain reaction (PCR) had the same results, B. melitensis biovar 3. Conclusion Brucella melitensis biovar 3 was identified as the pathogen of the three cases from Suizhou city and cases were mainly caused by direct contact with sick animals and their meat.

Objective To describe spatial distribution of brucellosis in Three-River Regions and research its epidemiological characteristics with the Geographic information technique. Methods We established Geographic Information System Database of brucellosis in Three-River Regions, epidemiological characteristics were overlay analyzed in infectious rate and prevalence rate with QHEndem ic-GIS and SPSS 17.0. Results Spatial characteristics of brucellosis were showed in GIS, the overlaid prevalence rates in Nangqian(2.00%/1.03%), Jiuzhi(0.81%/0.63%)and Dari(0.58%/0.33%)in inspection period were higher than that in inspection period. Overlaying the prevalence of brucellosis before and after inspection period, the prevalence show an upward trend. Conclusion GIS could reflect the state of brucellosis in Three-River Regions directly and correctly. Prevalence of brucellosis showed a sporadic phenomenon with spreading the neighborhood region, the prevalence showed an up-ward trend in these region, and provide scientific data of brucellosis prevention and control.

Objective To explore the feasibility of dilution rose bengal plate agglutination test (RBPT) for the diagnosis of brucellosis. Methods Serum samples from 93 brucellosis patients who came from Inner Mongolia, China in 2013 were tested by both dilution RBPT and standard tube agglutination test (SAT). According to the correlation between the results of the two tests and using the results of SAT as criteria, the sensitivity and specificity of RBPT were analyzed at different dilutions as cut-off values to evaluate the diagnostic accuracy. Results The area under the receiver operating characteristic curve for dilution RBPT was 0.946. When taking1∶2 as the cut-off value, the Youden index was 0.893, the sensitivity was 89.30% , and the specificity was 100%; when taking1∶4 as the cut-off value, the Youden index was 0.631, the sensitivity was 63.10%, and the specificity was 100%; when taking1∶8 as the cut-off value, the Youden index was 0.571, the sensitivity was 57.14%, and the specificity was 100%; when taking1∶16 as the cut-off value, the Youden index was 0.191, the sensitivity was 19.05%, and the specificity was 100%. Conclusion Dilution RBPT has the highest diagnostic accuracy at a dilution of1∶2, which provides a reference for the diagnosis of brucellosis.

【Abstract】 Objective To study the method to identify Brucella suis biovar 1 wild strain and S2 vaccine strain.  Methods A total of 21 strains of B.suis biovar 1 wild strain were analyzed by AMOS-PCR, PFGE and multiple-locus variable-number tandem-repeat analysis (MLVA).   Results B.suis biovar 1 and S2 could be discriminated by MLVA Bru9 and Bru16. Conclusion The MLVA assay can be applied to identification of wild strains and vaccine strain and can be proposed to as a complement of classical biotyping methods.

Objective Establishing the infective model of BALB/c and KM mice with Bartonella henselae in the lab, inspecting the dynamic variety of bacteremia and IgG andtibody in the mice. Methods BALB/c and KM mice were inoculated with B.henselae by routes intraperitoneally with a dose of 2.5×10 ⁸ cfu/ml, Enzyme-linked immunosorbent assay(ELISA) of serum samples collected as early as 2 d after infection indicated humoral immune responses to B.henselae. Results Specific humoral responses remained through 36 d. From the 6 d PI, the value of A begain to drive up, and the climax was in the 36 d. Culture analysis of blood from mice infected with B.henselae yielded positive results 2 d after infection, and the cultures was detected by polymerase chain reaction by 2 d and up to 36 d. Conclusion Analysis of samples revealed induction of B.henselae-specific immunoglobulin G from 6 to 36 d after infection.