[An immunofluorescence assay for the detection of SARS associated coronavirus antibody based on recombinant nucleocapsid antigen and its application].

OBJECTIVE: To establish a new technique for SARS-CoV antibody test to detect infection of severer acute respiratory syndrome (SARS). METHODS: Nucleocapsid gene was obtained by reverse transcription and polymerase chain reaction from a SARS patient and inserted into the vector pFastBacHTa expressing baculovirus. Insect Sf9 cells were transfected with the recombinant baculovirus expressing SARS nucleocapsid antigen and then cultured, fixed by acetone so as to make SARS-specific antigen. Immunofluorescence assay (IFA) technique and plaque reduction neutralization test (PRNT) were used to detect 7 samples of sera of 4 newly diagnosed SARS patients collected in different days, 48 samples of convalescent sera of SARS patients, 24 serum samples of healthy person undergoing physical examination, and 40 serum samples from non-SARS patients with fever by double blind test. RESULTS: The recombinant SARS-specific antigen reacted only with SARS positive sera but not with normal sera. Double blind test showed that 45 of the 46 PRNT positive sera were IFA positive with an accordance rate of 97.8%. 7 samples of sera from 4 SARS patients in acute progressive stage in Guangdong province were all IFA positive. SARS antibody could be detected since the sixth day after onset, and the titer increased from 1:40 to 1:600 on the ninth day. CONCLUSION: Immunofluorescence assay is highly specific and sensitive in detection of SARS. This reagent is safe and easy to prepare.

[Serological surveillance on vaccine against hemorrhagic fever with renal syndrome].

OBJECTIVE: To observe the serological and epidemiological efficacy of hemorrhagic fever renal syndrome (HFRS) vaccine in Zhejiang province. METHODS: Immunofluorescent antibody assay and Mcro-CPE method were used to test specific IgG antibody and the titer of neutralizing antibody. RESULTS: Two weeks after the injection of the third dose, the sero-conversion rates by both immunofluorescent antibody test (IgG) and neutralization test were 100.0% (67/67) (95% CI: 96.3 - 100.0) and 44.4% (8/18)(95% CI: 22.0 - 69.0) with geometric mean titers (GMTs) 72.1 and 4.6 respectively. The rates of seroconversion of immunofluorescent antibody by immunofluorescence antibody assay (IFA) were 28.6%, 83.3%, 75.0%, 53.1%, 22.6%, 10.0% and 55.0% before reinforcement, two weeks, one year, one year and a half years, two years, three years and five years after reinforcement. The rates of neutralizing antibody seroconversion by the Mcro-CPE method were found as 14.8%, 55.6%, 35.0%, 31.3%, 26.0%, 10.0% and 50.0% respectively. We found some antibody dependent immunization enhancement phenomenon among the inoculated population, but further observation was needed. CONCLUSION: HFRS vaccine was immunologically effective and the duration of serous antibody last long.