Analysis of monitoring results on plague in Ulanqab City of Inner Mongolia in 2018 / 中华地方病学杂志

Objective@#To analyze the plague monitoring results in Ulanqab City of Inner Mongolia in 2018, to master the changes in rat density and the prevalence of plague in rats, and provide a basis for scientific prevention and control of plague.@*Methods@#According to "The Plague Monitoring Scheme of Inner Mongolia", we surveyed Siziwang Banner, Chahar Right Back Banner, Huade County, and Shangdu County of Ulanqab City from April to November 2018 to monitor the plague. Rat density was surveyed using a one-day bow clamp method; small rodent was surveyed using a 5 m clamping method. Rodents were obtained by sample method, 5 m clamping method, daily method, collecting dead animals and the like, and fleas were picked up from the captured rats and rat nest. The rodents and fleas were carried out pathogen detection, the serum of rodents was tested by indirect hemagglutination test. Laboratory test results were analyzed based on the "Diagnostic Criteria for Plague" (WS 279-2008).@*Results@#Totally 1 463 mice were captured overlapping a monitored area of 416 hm2, the average rat density was 3.52 per hectare; the number of Meriones unguiculatus was 1 235, and the rat density was 2.97 per hectare. Totally 1 603 mice were grooming, 404 mice with fleas, the flea infected rate was 25.20%, the number of fleas were 1 348, and the flea index was 0.84. A total of 22 mouse nests were dug, 17 nests with fleas, the flea infected rate was 77.27%, the number of fleas were 131, and the flea index was 5.95. Totally 1 603 rodents were checked by etiology, the results showed that 7 plague rats were all Meriones unguiculatus. Totally 1 479 fleas of 581 groups were cultured, 14 fleas of 5 groups were detected, 3 fleas of 1 group in Siziwang Banner, and 11 fleas of 4 groups in Huade County. Totally 243 samples of murine animal serum were tested and the results were all negative.@*Conclusions@#The epidemic of plague in Ulanqab City is in an active state, so monitoring should be strengthened in this area to prevent the prevalence of human plague.

Analysis of monitoring results on plague in Ulanqab City of Inner Mongolia in 2018 / 中华地方病学杂志

Objective To analyze the plague monitoring results in Ulanqab City of Inner Mongolia in 2018,to master the changes in rat density and the prevalence of plague in rats,and provide a basis for scientific prevention and control of plague.Methods According to "The Plague Monitoring Scheme of Inner Mongolia",we surveyed Siziwang Banner,Chahar Right Back Banner,Huade County,and Shangdu County of Ulanqab City from April to November 2018 to monitor the plague.Rat density was surveyed using a one-day bow clamp method;small rodent was surveyed using a 5 m clamping method.Rodents were obtained by sample method,5 m clamping method,daily method,collecting dead animals and the like,and fleas were picked up from the captured rats and rat nest.The rodents and fleas were carried out pathogen detection,the serum of rodents was tested by indirect hemagglutination test.Laboratory test results were analyzed based on the "Diagnostic Criteria for Plague" (WS 279-2008).Results Totally 1 463 mice were captured overlapping a monitored area of 416 hm2,the average rat density was 3.52 per hectare;the number of Meriones unguiculatus was 1 235,and the rat density was 2.97 per hectare.Totally 1 603 mice were grooming,404 mice with fleas,the flea infected rate was 25.20％,the number of fleas were 1 348,and the flea index was 0.84.A total of 22 mouse nests were dug,17 nests with fleas,the flea infected rate was 77.27％,the number of fleas were 131,and the flea index was 5.95.Totally 1 603 rodents were checked by etiology,the results showed that 7 plague rats were all Meriones unguiculatus.Totally 1 479 fleas of 581 groups were cultured,14 fleas of 5 groups were detected,3 fleas of 1 group in Siziwang Banner,and 1 1 fleas of 4 groups in Huade County.Totally 243 samples of murine animal serum were tested and the results were all negative.Conclusions The epidemic of plague in Ulanqab City is in an active state,so monitoring should be strengthened in this area to prevent the prevalence of human plague.

Identifation methods of Brucella Melitensis / 中华地方病学杂志

Objective To establish genotyping methods for rapid identification of Brucella melitensis (B.melitensis) biovar 1,2 and 3 and to verify these method.Methods Single nucleotide polymorphism of RpoB gene and tandem repeat sequence (TRS) Bru42 of standard reference strain 16M were used to design primers,then the RpoB-PCR and TRS-PCR method were established for identification of B.melitensis standard reference strains,these two methods were used to identify clinical isolates of B.melitensis and compared with the conventional methods.Results The results of B.melitensis standard reference strains (biotype 1,2,3) identified by RpoB-PCR and TRS-PCR were consistent with those of the conventional identification methods.Totally 50 clinical isolates [including B.melitensis biovar 1 (17),2 (3) and 3 (30)] were identified as RpoB-2 genotype,only one B.melitensis biovar 1 strain was identified as RpoB-3 genotype.Genotype identification results of standard reference strains and clinical isolates with the same biotype were not exactly the same.Fothermore,TRS-PCR experiment displayed that 51 clinical isolates were all genotype 2 of B.melitensis (genotype TRS-2).Conclusions There is no clear relationship between biovars and genotypes within B.melitensis,and significant difference exists between B.melitensis standard reference strains and clinical isolates within RpoB gene.Bru42 can not be used for genotyping clinical isolates of B.melitensis.

Identifation methods of Brucella Melitensis / 中华地方病学杂志

Objective To establish genotyping methods for rapid identification of Brucella melitensis (B.melitensis) biovar 1,2 and 3 and to verify these method.Methods Single nucleotide polymorphism of RpoB gene and tandem repeat sequence (TRS) Bru42 of standard reference strain 16M were used to design primers,then the RpoB-PCR and TRS-PCR method were established for identification of B.melitensis standard reference strains,these two methods were used to identify clinical isolates of B.melitensis and compared with the conventional methods.Results The results of B.melitensis standard reference strains (biotype 1,2,3) identified by RpoB-PCR and TRS-PCR were consistent with those of the conventional identification methods.Totally 50 clinical isolates [including B.melitensis biovar 1 (17),2 (3) and 3 (30)] were identified as RpoB-2 genotype,only one B.melitensis biovar 1 strain was identified as RpoB-3 genotype.Genotype identification results of standard reference strains and clinical isolates with the same biotype were not exactly the same.Fothermore,TRS-PCR experiment displayed that 51 clinical isolates were all genotype 2 of B.melitensis (genotype TRS-2).Conclusions There is no clear relationship between biovars and genotypes within B.melitensis,and significant difference exists between B.melitensis standard reference strains and clinical isolates within RpoB gene.Bru42 can not be used for genotyping clinical isolates of B.melitensis.

Designing, Expression and Immunological Characterization of a Chimeric Protein of Mycoplasma pneumoniae.

Mycoplasma pneumoniae is thought to be the simplest and smallest cell wall-deficient bacterium which can cause chronic respiratory infections. Recently vaccination has been a possible and reliable way to reduce the spreading and infection effectively. In this study, the transmembrane proteins P116N (the N-terminal of P116), P1C (the C-terminal of P1), P30, and P116N-P1C-P30 (MP559 for short), a chimeric protein were expressed using prokaryotic expression system. The four purified recombinant proteins were synergized with freund's adjuvant and immunized New Zealand White rabbits respectively for three times. The IgG antibodies collected from immunized rabbits and mouse were analyzed by ELISA to analyze the immunogenicity and antigenicity. The results showed the four different recombinant proteins could induce strong humoral immune response. Protein MP559 could react with antisera from rabbit immunized with P1C, P30, and P116N, indicating MP559 was well designed and presented antigen epitopes of all the three antigens. Antibodies against P116N, P1C, and P30 could be stimulated by MP559 immunization, indicating MP559 has a potential to replace the three antigens as a vaccine candidate. This study laid a substantial foundation for the vaccine development of M. pneumoniae, and at the same time provided a essential strategy for the vaccine research of other pathogen.

Eucaryotic expression and bioactivity analysis of the recombinant HSV-gD1 / 生物工程学报

Envelope proteins of herpes simplex virus (HSV) plays a vital role not only in the infection process of adsorption and invasion but also in the stimulation to the organism that gives rise to immune response. Among the envelope proteins, glycoprotein D (gD), which can induce specific immune response, are the primary targets of humoral and cellular immunity of the host. In order to analyze the antigenicity and immunogenicity of HSV-gD1, we chemically synthesized the extracellular domain fragment gene of gD1, cloned it into eucaryotic expression vector pCEP4, and transfected the HEK293 cells with the recombinant vector. Then we identified the recombinant protein by Western blotting, and detected antigenicity of the protein by ELISA. Finally, we used the purified gD1 protein to immunize Kunming mice in 1, 3, 5 weeks, and collected antiserum in 3, 5 and 7 weeks. We titrated the sera for the detection of anti gD1 using an ELISA assay. Gene sequencing analysis demonstrated that the recombinant plasmid pCEP4-gD1 was constructed successfully. Western blotting analysis indicated one major protein band, which molecular weights is approximate 46 kDa corresponding to the truncated forms of gD1 protein, was observed. ELISA assay showed that the expressed recombinant protein gD1 had good antigenicity. After the third immunization, antibody titer of the mouse anti-gD1 was at least 5 x10(3). The successful expression of the recombinant protein gD1, which can induce humoral immune response, lays a foundation for serological diagnosis and vaccine study of HSV.