Objective To establish a CVS-11 pseudovirus particles ( pp)-based assay for detec-tion of neutralizing antibody against rabies virus. Methods An improved rapid fluorescence focus inhibition test ( RFFIT) for detection of neutralizing antibody against rabies virus ( RVNA) was established based on the CVS-11 pseudovirus expressing a luciferase reporter gene. Forty-six human serum samples were analyzed with the improved RFFIT and the results were compared with those by using standard RFFIT. Moreover, the improved RFFIT was used to detect the titers of RVNA in 91 serum samples collected from pet dogs and pet-breeders in Beijing. Results The coincidence rate of the improved RFFIT and the standard RFFIT was 100% regarding to the analysis of 46 human serum samples and 5 negative reference serum samples. Moreo-ver, the RVNA titers of all serum samples obtained with CVS-11 pseudovirus-based assay showed a signifi-cant high correlation with those obtained with standard RFFIT (n=46, r=0. 94, P<0. 000 1). All of the 91 serum samples collected from pet dogs and pet-breeders in Beijing were positive for RVNA as indicated by the improved RFFIT with a mean titer of 33. 01 IU/ml. Conclusion We established an improved RFFIT based on the CVS-11 pp expressing luciferase reporter gene, which might be used as a reliable alternative RFFIT for measuring RVNA titer. Analysis of the 91 serum samples collected in Beijing with the improved RFFIT showed that all samples were positive for RVNA.

Objective:To analyze the immune effect of tetanus toxoid (TT) and to provide reference for improving immunoprophylaxis strategies against tetanus.Methods:From 2019 to 2021, the TT-containing vaccine (TTCV) immunization history of patients treated for trauma in Luoshan Street Community Health Service Center of Jinjiang City were investigated. Serum tetanus antibody (TAB) levels were detected in 353 subjects (group A) 5-10 years after TTCV immunization, and the proportions of subjects with different TAB levels (<0.01 IU/ml, 0.01-0.10 IU/ml, >0.10 IU/ml) in different age groups were compared. Sixty-eight subjects (group B) aged 14-83 years with TTCV immunization history and TAB level of 0.01-0.10 IU/ml and 133 subjects (group C) aged 17-77 years without TTCV immunization history were inoculated with one dose and three doses of TT respectively, and the changes in TAB level were observed 28 d after immunization.Results:In group A, the proportions of subjects with different TAB levels in different age groups were statistically significant (χ 2=47.69, P<0.001). The proportions of subjects in which TAB levels were <0.01 IU/ml and 0.01-0.10 IU/ml increased with age. In group B, 66 out of the 68 subjects had TAB >0.10 IU/ml 28 d after one dose of TT immunization. There were statistically significant differences in the proportions of subjects whose TAB levels were 0.01-0.10 IU/ml and >0.10 IU/ml before and after TT immunization (χ 2=128.23, P<0.001). In group C, before three doses of TT immunization, 129 patients had TAB <0.01 IU/ml and four patients had TAB in the range of 0.01-0.10 IU/ml; 28 d after three doses of TT immunization, only one case had TAB in the range of 0.01-0.10 IU/ml and 132 cases had TAB >0.10 IU/ml. The proportions of group C subjects with different TAB levels before and after TT immunization were statistically significant (χ 2=262.80, P<0.001). Conclusions:Five years after TTCV immunization, the proportions of individuals with TAB <0.01 IU/ml and in the range of 0.01-0.10 IU/ml increased with age. For people without TTCV immunization history and those with decreased TAB protection after TTCV immunization, strengthening TT immunization could significantly improve the TAB protection.

Rabies is one of the important zoonotic infectious diseases, with a mortality rate of almost 100%. Rabies is a vaccine preventable disease, and proper rabies exposure prophylaxis can effectively prevent the occurrence of human rabies. In recent years, there has been significant progress in clinical research on the rabies exposure prophylaxis both domestically and internationally. World Health Organization (WHO) released the Rabies Vaccine: WHO Position Paper-April 2018. In order to guide medical institutions of all levels in rabies exposure prophylaxis, the National Administration of Disease Prevention and Control, in conjunction with the National Health Commission of the People's Republic of China, organized the Rabies Vaccine Working Group of the National Immunization Program Technical Working Group and invited experts to revise and issue the National Regulation for the Rabies Exposure Prophylaxis (2023 Edition). This article compares the National Regulation for the Rabies Exposure Prophylaxis (2009 Edition) and interprets the updated key points and supporting basis of the new version of the guidelines to guide clinical application and implementation.

Objective:To evaluate the immunogenicity and immune persistence of purified Vero cells rabies vaccine (PVRV) with Zagreb and Essen regimen.Methods:Prospective study: Patients with first Class II exposure to rabies were recruited from the Dog Injury Cinic of Guangxi Center for Disease Control and Prevention (Guangxi CDC) and randomly divided into the Zagreb (2-1-1) and Essen (1-1-1-1-1) regimen group. All patients were inoculated with the vaccines from the same manufacturer and batch, and 3 ml serum was collected at the 45th day and in 1-year, 2-year and 3-year after immunization. Rapid fluorescent inhibition test (RFFIT) was used to detect rabies virus neutralizing antibody (RVNA). The attenuation of RVNA positive rate and geometric mean titer (GMT) with time was analyzed. Retrospective study: The informed consents for rabies vaccine in the Dog Injury Clinic of Guangxi CDC were checked out. The patients who were injected with PVRV (same manufacturer but unlimited batch) but without passive immune agents for the first time within 3 years were selected and divided into 1-year, 2-year and 3-year group. Each group was further divided into Zagreb and Essen regimen group. The serum (3 ml) was collected at 1 year, 2 years and 3 years after immunization and detected the RVNA by RFFIT.Results:Prospective study: The RVNA positive rates on the 45th day and in 1-year, 2-year and 3-year after immunizationin in the Zagreb and Essen regimen group were 100%, 95%, 85%, 80% and 98.25%, 89.47%, 89.47%, 85.96%, respectively. There was no statistically significant difference in the RVNA positive rates at the same time point between the two regimen groups ( P>0.05). The RVNA GMT on the 45th day and in 1-year, 2-year, and 3-year in the Zagreb and Essen regimen group were 11.32 IU/ml, 1.69 IU/ml, 1.30 IU/ml, 1.30 IU/m and 13.18 IU/ml, 2.13 IU/ml, 1.87 IU/ml, 1.84 IU/m, respectively. There was no significant difference in the RVNA GMT levels at the same time point between the two regimen groups ( F=1.971, P=0.164). The RVNA GMT levels in the two regimen groups had the same trend of attenuation over time (time*group F=0.702, P=0.435). Retrospective study: The RVNA positive rates in 1-year, 2-year and 3-year after immunization in the Zagreb and Essen regimen group were 100%, 95%, 91.43%和94.73%, 86.21%, 87.5%, respectively. There was no statistically significant difference in the RVNA positive rates at the same time point between the two regimen groups ( P>0.05). The RVNA GMT in 1-year, 2-year, and 3-year groups after immunization in the Zagreb and Essen regimen group were 2.65 IU/ml, 2.03 IU/ml, 1.57 IU/ml和3.2 IU/ml, 2.58 IU/ml, 2.45 IU/ml, respectively. There was no significant difference in the RVNA GMT levels at the same time point between the two regimen groups ( P>0.05). Conclusions:The PVRV showed the same excellent immunogenicity and immune persistence after the vaccination with the Zagreb and Essen regimens.

Non-neonatal tetanus is an acute, specific, toxic disease in patients over 28 days of age, characterized by continuous rigidity and paroxysmal spasms of the skeletal muscles throughout the body caused by the intrusion of Clostridium tetani through skin or mucosal membrane into the body and reproducing in anaerobic environments to produce exotoxins. The mortality rate of severe patients is close to 100% without medical intervention. Even with aggressive comprehensive treatment, the global mortality rate remains at 30%-50%, making it a potentially fatal disease. In order to standardize the diagnosis, treatment and prevention of non-neonatal tetanus, based on "Regulation for Diagnosis and Treatment of Non-neonatal Tetanus (2019 Edition)", experts have revised this regulation according to clinical practice and recent research progress in this field to guide medical institutions in the prevention and control of non-neonatal tetanus. This article interprets the key points and basis for updating the 2024 edition regulation to guide clinical implementation and application.

The situation of prevention of non-neonatal tetanus in China is severe. Strengthening the active immunization with tetanus toxoid vaccine (TTCV) is the key to prevent the non-neonatal tetanus. Through the detection of tetanus antibody (TAB), the immune status of individual can be determined, so as to implement the active immunization of TTCV correctly. The research on TAB detection technology is stagnant in aboard, but still in a development process in China since there is a realistic demand for TAB detection. This review collects relatively limited data of TAB detection technology in China, and summarizes the techniques such as mice toxin neutralization test (MTNT), indirect hemagglutination assay (IHA), double agar gel immune diffusion test (Rubin method), enzyme-linked immunosorbent assay (ELISA) and colloidal gold (CG), in order to provide a comprehensive basis for domestic TAB detection. The TAB detection technology in China has not yet achieved international recognition due to the lack of comparative study of domestic and international institutions and reference reagents. The special domestic situation of tetanus prevention makes the research of TAB detection technology have a certain practical significance, and rapid detection reagents such as ELISA and CG method have a certain application value in China.

Objective To select and identify the cell line for detecting Enterovirus 71 (EV71).Methods pWSK-T7-EV71-GFP containing green fluorescent protein (GFP) gene is an infectious clone for EV71,based on which the UGFP cassette was constructed by inserting GFP gene into 5' and 3' untranslated region (UTR) of the genome of EV71.The lentiviral expression plasmid pLV-UGFP containing UGFP was constructed on the basis of pLV-Puro,a lentiviral vector.To obtain lentivirus,pLV-UGFP plasmids were transfected together with the packaging plasmids into HEK293T cells by liposomes.Then,the target cells BHK-21 were infected with the lentivirus particles.Puromycin-resistant cell colonies were detached from the 6 well-plate and sub-cloned by use of 96 well-plate.Finally,we selected the packaging cell lines that could express the defective replicons stably,named BHK/UGFP cells.Results GFP expression assays indicated that BHK/UGFP cells infected with EV71 could express GFP at 48 hours post-infection,while no green fluorescence was observed after BHK/UGFP cells were infection with Sindbis virus (SINV,XJ-160) or Japanese encephalitis virus (JEV,P3),demonstrating that the selected cells could specifically detect EV71 infection.The sensitive assay results indicated that this method on the basis of BHK/UGFP cells could at least detect 10 PFU/ml of EV71 in tissue culture.Conclusion This result indicated that the BHK/UGFP cells selected in this study were specific and effective to detect EV71 from tissue culture.

Human vaccine is the most effective tool to prevent infectious diseases. At present, there are dozens of vaccines that can prevent dozens of infectious diseases. Vaccines for human use include vaccines that do not contain living micro-organisms (inactivated vaccines, toxoid vaccines, component vaccines), and vaccines that contain living micro-organisms (live attenuated vaccines and vector vaccines). The research and development of human vaccine includes clinical trial, process development and test method research, and the production process of human vaccine is very important for the preparation of high-quality and reliable vaccine. In this paper, the development of human vaccines, the production process and the characteristics of various human vaccines are reviewed.

Rabies is a zoonotic infectious disease caused by lyssavirus and characterized by central nervous system symptoms. The fatality rate of rabies is almost 100%. About 59 000 cases die of rabies worldwide every year, mainly in Asia and Africa. China is an epidemic country of rabies. Grade II and III exposures are the main types of rabies exposures in China. Standardized post?exposure prophylaxis (PEP) can prevent rabies almost 100%. Human Rabies Vaccine Technical Working Group, National Immunization Advisory Committee and invited experts reached an expert consensus on PEP by referring to the World Health Organization′s position paper on rabies vaccine in 2018 and related research progress in recent.

Human vaccine is the most effective tool to prevent infectious diseases. At present, there are dozens of vaccines that can prevent dozens of infectious diseases. Vaccines for human use include vaccines that do not contain living micro-organisms (inactivated vaccines, toxoid vaccines, component vaccines), and vaccines that contain living micro-organisms (live attenuated vaccines and vector vaccines). The research and development of human vaccine includes clinical trial, process development and test method research, and the production process of human vaccine is very important for the preparation of high-quality and reliable vaccine. In this paper, the development of human vaccines, the production process and the characteristics of various human vaccines are reviewed.