1.A Semi-quantitative Serological Method to Assess the Potency of Inactivated Rabies Vaccine for Veterinary Use
Ye LIU ; Shoufeng ZHANG ; Fei ZHANG ; Rongliang HU
Virologica Sinica 2012;27(4):259-264
Potency is one of the most important indexes of inactivated vaccines.A number of methods have been established to assay the potency,of which the NIH test and single-dose mouse protection test are the prescribed methods.Here,we report a method to semi-quantitatively assay the potency of an inactivated rabies vaccine,which uses fewer animals and takes less time to complete.Depending on the quality requirements of a vaccine(e.g.minimum potency),a rabies reference vaccine is,for example,diluted to the minimum potency,and 50 μL of the dilution is taken to inoculate 10 mice.The same amount of the test rabies vaccine is inoculated into another 10 mice.After two weeks,all mice are bled and serum samples are assayed for viral neutralizing antibody by the fluorescent antibody virus neutralization(FAVN) test.By comparing the median and interquartile range of antibody titers of the reference vaccine with those of the test vaccine,the test vaccine potency can be semi-quantitatively judged as to whether it is in accord with the required quality.The reliability of this method was also confirmed in dogs.The procedure can be recommended for batch potency testing during inactivated rabies vaccine production.
2.Studies of Di-n-butyl Phthalate-OP Emulsion in the Treatment of Demodicidosis
Hui XIA ; Shoufeng HU ; Weiju MA ; Jihua GE
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
Objective To observe the curative effect of Di\|n\|butyl phthalate\|OP emulsion in the treatment of demodicidosis. \ Methods\ 447 cases with Demodex infection on face were treated with Di\|n\|butyl phthalate\|OP emulsion. Among them, 30 cases suffering from acne, tetter and pustule were also randomly observed. 20 days after treatment negative conversion rate and the therapeutic effect were evaluated. At the same time, the effect of this solution was compared with that of other three medicaments (FuManLing, 2% metronidazole and 8% metronidazole preparations). In vitro test of mites\|killing, toxicity test in experimental animals and the safety evaluation for local application were also performed. \ Results \ Results showed that the negative conversion rate was 92\^8%(415/447), effective rate for the cases showing evident face damage was 90\^0%(27/30). The result also indicated that the OP emulsion medicament was more effective than other three medicaments (P
3.Survey and Treatment of Pulmonary Acariasis among the Workers Involving inTraditional Chinese Medicinal Materials
Hui XIA ; Shoufeng HU ; Xingbao CHEN ; Xiuying RU ; Xingyi SHAO
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(02):-
Objective To investigate the prevalence of pulmonary acariasis among the employees working on traditional Chinese medicinal materials and observe the effect of treatment. Methods History inquiry, detection of mites in sputum, blood examination for eosinophils and specific antibodies, x-ray chest film were carried out for 327 workers involving in traditional Chinese medicinal materials. Mites were found in sputum in 121 persons who were then treated with metronidazole, twice a day with a daily dosage of 0.8g for seven days as a course of treatment. Two courses were conducted with an interval of 7-10 day. Prevalence and morbidity in different groups of occupation, age, and sex were analyzed. Results The overall infection rate of mites in sputum was 37.0% (121/327) with an average morbidity of 12.5% (41/327). Among the four types of worker investigated, the highest infection rate (51.8%), and morbidity (18.6%) were in those working in transfer warehouse; the second highest infection rate (40.7%) and morbidity (15.7%) were in employees in factory of Chinese traditional medicine. Both groups showed a significant difference with others(?2inf=11.36,P0.05). After treatment with metronidazole, 88.4% showed negative in sputum examination for mites and the efficacy of the treatment for pulmonary acariasis was 92.3%. Conclusions Employees engaged in traditional Chinese medicinal materials are one of the groups at the highest risk of pulmonary acariasis. Metronidazole is effective in treating the infection.
4.The effects treated with PICKCa rabies vaccine to mice and beagle dogs infected rabies wild viruses and volunteers immunized with PICKCa rabies vaccine
Haixiang LIN ; Gelin XU ; Jie WU ; Shoufeng ZHANG ; Rongliang HU ; Lietao LI ; Fang LIU ; Shirui ZHANG ; Yi ZHANG
Chinese Journal of Experimental and Clinical Virology 2016;(1):37-40
Objective To test the immune therapeutic effects of PICKCa adjuvant rabies vaccine for the post-exposure mice and beagle dogs and immune memory effect for human.Methods For the mice and beagle dogs first to infect with rabies wild virus and then immunize with PICKCa rabies vaccine.For volunteers, first to immunize with PICKCa rabies vaccine after 2.5 years to take peripheral blood mononuclear cell and then to check cells secreted interferon-gwith flow cytometry. Results In 3 independent assays of post-explosure immunization of mice and beagle dogs PICKCa rabies vaccine were much better then commercial adjuvant-free rabies vaccines, the protective rate from 20%-30%to 70%-100%Also, in the assay of cells secreted IFN-γby peripheral blood mononuclear cells from volunteers immunized by PICKCa rabies vaccine the IFN-γcell levels were much higher than control.Conclusions Compared with commercial adjuvant-free rabies vaccines the PICKCa rabies vaccine had significant immune therapeutic effects in the mice and dogs.Also PICKCa rabies vaccine had good immune memory effect in human.
5.Establishment and evaluation of a method for detection of ASFV antigen by doub-le-antibody sandwich ELISA
Qixuan LI ; Huixian YUE ; Yiqian JIANG ; Yanyan ZHANG ; Teng CHEN ; Shuchao WANG ; Shoufeng ZHANG ; Rongliang HU
Chinese Journal of Veterinary Science 2024;44(8):1579-1584,1592
African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10%.The total co-incidence rate with real-time fluorescence quantitative PCR was 92%(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.
6.Construction and immunogenicity study of 293T cell line stably expressing rabies virus glycoprotein
Yu WANG ; Shoufeng ZHANG ; Tingting YU ; Lijuan MI ; Naiquan YAO ; Faming MIAO ; Rongliang HU
Chinese Journal of Veterinary Science 2024;44(11):2393-2399
In order to obtain a cell line which high expressed of rabies virus glycoprotein the produc-tion of rabies subunit vaccine,the RABV glycoprotein gene sequence was amplified by RT-PCR and cloned into the lentiviral expression vector.The recombinant plasmid pLV-RVG,envelope plasmid pMD2.0G and packaging plasmid psPAX2 were co-transfected into HEK-293T cells to harvest lentivirus and infect new HEK-293T cells.The 293T-RVG cell line was obtained by puro-mycin pressure screening.Indirect immunofluorescence assay(IFA)and Western blot showed that the 67 kD RABV-G protein was highly expressed in the cells.The neutralizing antibody titer reached 2.19 IU/mL within 14 days after immunization,which was higher than the internationally recognized protection threshold(0.5 IU/mL).This study showed that the cell line 293T-RVG with stable and high expression of rabies virus glycoprotein was successfully constructed.The cell line could protect mice against rabies virus challenge and could be used as a subunit vaccine with poten-tial for large-scale production and application.