1.Preparation of mouse anti-recombinant SAG1 antigen monoclonal antibody of Toxoplasma gondii by intrasplenic immunization
Nan WU ; Danqiu ZHOU ; Wei RUAN ; Ligui WU ; Huizhang ZHANG
Fudan University Journal of Medical Sciences 2010;37(2):184-188
Objective To prepare monoclonal antibody in mice so as to develop an ELISA method for diagnosis of Toxoplasma gondii infection during the initial stage. Methods The mice were immunized by combining routine and intrasplenic immunization with recombinant SGA1 antigen. B lymphocyte hybridization technique was applied to prepare the anti-SAG1 McAbs. Positive clones were screened using ELISA and subcloned to establish cell lines. Ascites was induced to produce the McAbs. Then the McAbs were purified by protein G chromatograph column. The specificity of McAbs was identified by Western blot and sandwich-ELISA. Sensitivity of the McAbs was determined using sandwich-ELISA. Comparasion was carried out between PCR and sandwich-ELISA method. Results Two positive clones were obtained and named as 3B6, 10C4, both could identify the native and recombinant SAG1 antigens. The sensitivity of 3B6, 10C4 was 31.3 ng and 62.5 ng, respectively. There was no cross reaction between the McAbs and positive sera from patients with schistosomiasis, ancylostomiasis or malaria. By using PCR and ELISA, the positive infection rate of T. gondii was 63.2% and 47.4%, respectively. Conclusions Therefore, mouse anti-rSAG1 antigen McAbs have been prepared successfully and primarily applied to early stage diagnosis of T. gondii infection.
2.Study on the relationship between HLA-B * 5801 and severe cutaneous adverse reactions caused by allopurinol
Xinju ZHANG ; Jiong ZHANG ; Weizhe MA ; Danqiu ZHOU ; Ming GUAN ; Hejian ZOU
Chinese Journal of Rheumatology 2014;18(4):259-262
Objective To investigate the relationship between HLA-B * 5801 allele and severe cutaneous adverse reactions caused by allopurinol or other drugs.The clinical value of HLA-B * 5801 as the marker of allopurinol-SCAR was evaluated.Methods Forty-three patients with allopurinol-SCAR,133 patients without SCAR after taking allopurinol for 3 months were included.Ninety-six patients with SCAR caused by other drugs and 148 healthy individuals were enrolled into the present study.HLA-B * 5801 allele was detected by PCR-SSP method.Data were analyzed by chi-square test.Results HLA-B * 5801 was present in 40 of 43 (93.0%) patients with allopurinol-SCAR,which was significantly higher than 19 of 148 (12.8%) in healthy subjects (x2=100.353,P<0.01,OR=90.5,95%CI 25.5-321.8).But there were no significant differences between allopurinol-tolerant patients and healthy controls(10 of 133,7.5,x2=2.141,P>0.05,OR=0.6,95%CI 0.2-1.2).And there were only 14 of 96 (7.5%) patients with SCAR caused by other drugs had HLA-B * 5801 (x2=0.152,P>0.05,OR=1.2,95%CI 0.6-2.4).Conclusion The study indicates that people with HLA-B * 5801 have a high risk of allopurinol-SCAR.HLA-B * 5801 is a specific and predictive marker for guiding the selection of uric acid lowing drug allopurinol.