Diagnostic Value Of RSJ26-SJ32 For Acute Schistosomiasis Japonicum

ObjectiveTo study the diagnostic value of rSj26-Sj32 for the acute schistosomiasis japonica. And to investigate the value of amplification Sj26, Sj32 and Sj14-3-3 encoding gene by PCR or RT-PCR for diagnosis the patients with acute schistosomiasis japonica.MethodsThe rSj26-Sj32 were purified by Ni-NTA kits from BL21(pET28α-Sj26-Sj32), and were identified by SDS-PAGE and western-blot. To detect the IgG or the IgM in sera of acute schistosomiasis japonicum by ELISA, Dot-ELISA and Dipstick, which were built by the purified rSj26-Sj32 protein and Schistosoma japonicum adult worm antigen(SjAWA) respectively, compared the patients with clonorchiasis sinensis, paragonimiasis westermani , alveolar echinococcosis, cystic echinococcosis,hepatitis B, lung tuberculosis and the health donors as the control groups.The antigen encoding gene of Sj26,Sj32 and Sj14-3-3 were amplified by PCR or RT-PCR from the DNA or the total RNA and identified by 1.2% AgaroseⅡ. At the same time, the DNA or the total RNA were obtained from the sera of patients with paragonimiasis westermani, clonorciasis sinensis and the normal subjects as the control groups.ResultIt was successful to obtain the rSj26-Sj32. The sensitivity and specialty were 98.00% and 97.67% in the acute schistosomiasis japonicum by rSj26-Sj32-IgM-ELISA, and 96.00% and 97.67% by SjAWA. There were various cross-reactivity with paragonimiasis and clonorchiasis by SjAWA-IgM-ELISA; but there were no cross-reactivity by rSj26-Sj32-IgM- ELISA.The sensitivity and specialty were 90.00%and 97.67% in acute schistosomiasis japonica with rSj26-Sj32-IgG-ELISA, but 92.00%and 97.67% with SjAWA. The cross reaction was 20.00% in patients with alveolar echinococcosis with SjAWA-IgG-ELISA, but no cross reaction with rSj26-Sj32.It showed that the sensitivity and specialty were 100.00%and 95.35% in acute schistosomiasis japonica with rSj26-Sj32-HRP-IgG-Dot-ELISA, but 100.00%and 93.02% with SjAWA. Moreover, cross reaction were emerged among sera from patients with clonorchiasis, paragonimiasis westermani, and alveolar echinococcosis between two methods.The sensitivity and specialty were 96.00%and 97.67% in acute schistosomiasis japonica with rSj26-Sj32-Immunogold-IgG-Dipstick, but 100.00%and 95.35% with SjAWA. Some cross reaction were emerged in paragonimiasis sinensis, clonorchiasis westermani and alveolar echinococcosis with SjAWA, but not with rSj26-Sj32.A 400bp Sj14-3-3 encoding gene was obtained by PCR or RT-PCR from the 50 schistosomiasis japonica patients, but the encoding gene of Sj26 and Sj32 weren't obtained, the control groups were negative. The sensitivity and specialty were both 100% by PCR or RT-PCR with Sj14-3-3. There was no cross reaction with patients with clonorchiasis sinensis or paragonimiasis westermani.Conclusion1. The rSj26-Sj32 might be used for the immunodiagnosis of acute schistosomiasis japonica.2. Amplification Sj14-3-3 coding gene by PCR or RT-PCR may be used for the gene diagnosis with the acute schistosomiasis japonicum.