Objective To investigate the rodents species diversity along the border area of China and Kazakhstan, and provide supporting data for the surveillance and control of rodent borne diseases. Methods Rodents were collected with trapping method at seven counties/districts and area in Yili, Boertala, Tacheng and Karamay. Rodents species were then identified with DNA barcodes. Results Totally 174 samples of small mammal were collected from three kinds of habitats, including 16 species of rodents belonging to Muridae, Cricetidae, Dipodidae, Gliridae and Sciuridae, and one shrew species of Soricidae. With the DNA barcoding method, samples belonging to two vole species(Microtus arvalis and M. gregalis) with similar morphologic characters were identified correctly. Gerbil samples recognized morphologically as Meriones meridianus were different genetically from M. meridianus from Eastern China. Conclusion High diversity of rodent species were found in alpine forests meadows and desert steppe habitats. The genetic distances between M. meridianus from northern Xinjiang and other areas of China are so profound that suggests there are cryptic species.

Objective To provide a preliminary evaluation of the self?made horseradish peroxidase (HRP) marked Hantavirus (HV) recombinant N protein (rNP) rNP?IgM direct capture ELISA diagnostic kit. Methods Assessment of the specificity and stability of the kit and comparison of clinical results with similar products were performed to evaluate the sensitivity of the kit in the detection of serum anti?HV?IgM. Results (1) The kit was only responsive to anti?HV?IgM positive serum, and irresponsive to anti?varicella?zoster virus?IgM (anti?VZV?IgM), anti?Japanese encephalitis virus?IgM (anti?JEV?IgM), anti?dengue virus?IgM (anti?DV?IgM), anti?hand, foot and mouth EV71 virus?IgM (anti?EV71?IgM) positive sera. No obvious reduction in serum anti?HV?IgM detecting capability was noticed after placement at 37 ℃ for 3 d. (2) In 144 sera samples in 120 patients with suspected hemorrhagic fever with renal syndrome, inconsistency was observed only in the anti?HV?IgM test results in 12 sera of 12 patients between the two kinds of kits, in which 8 primary sera samples were detected positive by the kit and negative by commercial ones (the secondary sera samples were positive for both kits); 3 primary samples (the secondary samples unavailable) were at the critical value for the kit and negative for commercial kits. Meanwhile, one other primary serum sample was positive for the kit and negative for the commercial ones (the secondary and tertiary ones positive for both). Conclusion The laboratory developed capture ELISA anti?HV?IgM diagnostic kits had favorable specificity, stability and sensitivity, suitable for the clinical diagnosis and epidemiological surveillance of HV infections.

【Abstract】 Objective To clarify the natural infection situation of rodents by Hantavirus (HV) and HV strains in Zhejiang province in 2007，and provide science evidence for the prevention and control of hemorrhagic fever with renal syndrome (HFRS)．Methods Lung tissue and serum from rodent were sampled, and IgG antibody from serum was tested by indirect immunofluorscence assay and direct immunofluorscence assay was adopted to test HV antigens from lung tissues. HV was isolated by Vero?E6 cells, and HV strains were identified by Anti?McAb．Results A total of 1129 rodents were captured in 2007．The dominant specie was Apodemus agrarius, and the positive rate of HV antigen in rodent lungs was 3.0%. The IgG antibody of 57 blood samples was positive, and the positive rate was 8.0%.  Six  strains  of  hantaan (HTN)  virus  were  isolated,  five  strains  from A.agrarius  and  one  from  Rattus norvegicus.  Conclusion There  are  natural  foci  of  HFRS  which  main  infection  sources  are A.agrarius in Zhejiang province, and HTN strain could be the main prevalence strains of HV.

Objective Vero cell-derived HFRS purified vaccine researching and producing. Methods The vaccinal strains Z ₁₀(type Ⅰ)and Z ₃₇(type Ⅱ)of HFRS bivalent inactivated vaccine were adapted in vero cells and continuously porpagated. The Vero-cell derived HFRS purified vaccine were developed using the different passages of these 2 viral strains.Results The strains Z ₁₀and Z ₃₇were adapted to Vero cells and grew on cells with high titers only after one time passages. The viral titers reached and maintained stably between 6.25-6.75and6.50-7.00logTCID ₅₀/ml respectively after serial passages in Vero cells. The amounts of virus antigen and viral titers of Vero cell-derived HFRS inactivated vaccine made from different passages of these viral strains were much higher than the former.The RPHI titers of strainsZ ₁₀increased from negative for Cp ₁to 1∶64for Cp ₁₀,and that of strains Z ₃₇from1∶2for Cp ₁to1∶64for Cp ₁₀. The viral titers (logTCID ₅₀/ml) of strains Z ₁₀raised from 4.25 for Cp ₁to 6.50 for Cp ₁₀and that of strains Z ₃₇from 6.25 for Cp ₁to 7.50 for Cp ₁₀. The viral titers and the amounts of virus antigen were up to the standand of biological products in China. Conclusion In the study on the HFRS purified vaccine in Vero cells,the most important thing is the passage adaptability of vaccinal strain in Vero cells.

Objective To select the vaccine strain of hantanvirus and the cell for inculation,the immunogenicity of hantanvirus and virus titer are tested.To establish the technics of vaccine development,the vaccine,the inactived adjuvant that do not weaken the hemoglutinin of hantanvirus is selected.To develop vaccine which has low aside effect and good immunogencity preventing clinical infection of hemmorrhagic fever with renal syndrome(HFRS).Methods By immunogenicity and pretentive effect test of animal,the vaccine strain is selected from isolates of hantanvirus;In comparison with formalin,the inactive effect is observed and used as guideline;To select the cell for vaccine development,the virus titer in cell,the time that cell welt and the times of harvest are observed.Results The Z10 strain(hantaan virus) and Z37 strain(Seoul virus) of hantanvirus is selected as vaccine strain from 45 strains.Both of them have good immunogenicity and high virus titer;βpropiolactone as inactived adjuvant does not weaken the hemagglutination antigen of vaccine and have low aside effect;The primary cell of merones unguiculatus kidney tissue as virus inculation cell have high virus titer and welt well.It can also havest virus many times.Conclusion Momovalent inactive vaccine(type Ⅰ,Ⅱ)and duvalent inactive vaccine against HFRS which has been developed by Zhejiang province center for disease control and preventive have low aside effect and good immunity.All of them can can protect clinical infection of HFRS and is worthwhile to used.

Objective Several vaccine candidate strains of hemorrhagic fever with renal syndrome(HFRS) were isolated from HFRS epidemic area,the sero type were identified and the antigenicity were studied.Methods The virus strains were continuously propagated in brain of sucking Mongolian gerbil.The characteristics of virus multiplication,viral titers and the amounts of virus antigen after serial passages were studied.Results 9 strains virus were isolated successfully from 32 positive mouse lung,of which 6 strains were determined as hantaan virus by Anti HFRS McAb and 3 strains as seoul virus.Virus titers and antigen titers of ZJ7(Hantaan virus,HTN) can reach 6.50 lgCCID ₅₀/ml、 1∶5 120,ZJ5(Seoul virus,SEO) and 6.00 lgCCID ₅₀/ml、 1∶5 120,respectively.Conclusion These two virus strain possessed high titers and good antigen and can be vaccine candidate strain of HFRS.

Objective To observe the adaptability of Hantavirus and the rule of these strains replication in Vero cells.Methods Virus are extracted every 3 days from Vero cells culture Hantavirus infected.Cytopathogenic effect(CPE) are observed until these cells degrade.Viral titers and the amounts of virus antigen after serial passages are observed by IFAT and RPHA.Results There was no cytopathogenic effect(CPE) in Vero cells after 32 days.Viral titers and the amounts of virus antigen reach to the top at 8-11th day after Hantavirus infect Vero cells.Viral titers and the amounts of virus antigen still keep up at the level of 5.00 Lg TCID ₅₀/ml and 1∶20 at 32th day after infaction.Conclusion The results suggest that these 2 candidate strains had adapted to Vero cells possessed high titers and good antigenicity and be feasible to prepare the HFRS vaccine in Vero cells.

Objective To study the epidemic characteristics and make the preventive strategy of HFRS.Methods FAT was used to detected HV antigen in lunges of mouse,and IFAT for antibody detection in HFRS patients.Results All 1 499 cases of whole province were mainly distributed in Shaoxing,Quzhou,Ningbo,Lishui,Taizhou of Zhejiang province.Summer and autum,winter were the two peak periods of HFRS in Zhejiang.Most of the cases were young strong peasants.Conclusion We should select bivalent vaccine to decreasing the incidence of HFRS in the mixed type areas where Apodemus agrarius is the main mouse.

Objective:To control morbidity of HFRS in high epidemic area.Methods:The integrated control measures were carried.The main way is to vaccinate people.The second way is to eliminate rat and health education.Result:The vaccine inculatory rate in the crowd from sixteen to sixty was 51.86%.Immune cucceed rate was 97.6%.From 1997 to 1999,the morbidity of HFRS in five villages decrease from 490.23 per 100000 to zero.Conclusion:The measures of integrated coutrol HFRS high epidemic area is efficient to reduce HFRS'S morbidity.

Objective:To observe the neutralizing antibody reaction after secondary immunization with Bivalent inactivated vaccine for epidemic hemorrhagic fever.Methods:Some volunteers were injected with bivalent inactivated vaccines for epidemic hemorrhagic fever(HFRS).3 doses were for primary immunization and one dose for secondary immunization after one and half year's interval.Neutralizing antibodies(NA)against HTNV and SEOV in sera before and after secondary immunization were detected by a plaque reduction neutralization test(PRNT).Result:TheGMT of NA against HTNV and NA against SEOV after secondary immunization were 1:28.29 and 1:32.49,respectively.The GMT of NA increased(3-10 fold)compared with that before secondary immunization.

A rapid fluorescent focus inhibition test of hemorrhagic fever with renal synidrom virus (HFRSV) has been established.All four HFRSV strains tested could form fluorescent focus on Vero-E cell culture.A linear re1ationshlp between the number of nuorescent focus and virus concentration was observed.By comparative testing of 20 samples of sera, it was proved that sensitivity of detecting neutralizing antibodies to HFRSV by RFFIT was sirnilar to that by PRNT.However, this method is easier, faster and more specific than the others.

This paper describes that the inactivated HFRS vaccine (type I） cultured in M.unguiculatus kidney cell was used on human, the induced neutralization antibody were observed Persistently.The results showed that about 90% seroconversion rate of neutralization antibody occurred after primary injection three times.But the persistent time was shorter.It reduced 30% in a year after primary immunization.When the secondary immunizatiom (one injection) was done, the persistent time was longer.About 50% seroconversion rate of neutralization antibody occurred at six years after secondary immunization.There fore, this suggestes that the secondary immunization be necessary.