Application of signaling protein 14-3-3 and 26 kDa glutathione-S-transferase to serological diagnosis of Schistosomiasis japonica.
Schistosomiasis japonica is currently one of the most serious parasitic diseases and over 670000 people are infected in China by the end of 2006. In order to establish an effective diagnostic method, the gene coding for Sj14-3-3 and Sj26kDa GST were cloned and expressed separately in Escherichia coli as fusion protein with His-tag. The rSj14-3-3 and 26kDa rSjGST were combinedly used as antigens for enzyme-linked immunosorbent assays (ELISA) to diagnose acute and chronic S. japonica. Our results showed that the sensitivity in diagnoses of both acute and chronic schistosomiasis was 94.4% (67/71) and 80.7% (96/119), respectively. The specificity was 94.7% applying 132 sera from people living in S. japonicum-free areas. The data also showed that the recombinant proteins cross-react with Clonorchis sinensis and hookworms at a rate of 11.8% and 5.3% respectively. Parallel tests were conducted among ELISA, indirect hemagglutination assay (IHA) and circular ovum precipitin test (COPT) to determine anti-S. japonicum antibodies in sera of patients with schistosomiasis, healthy control, and those infected with other parasites and the results showed no significant difference in sensitivity for acute schistosomiasis between ELISA and IHA assays (chi(2)=1.33, P>0.05), but significant between ELISA and COPT assays (chi(2)=6.72, P<0.01). Our results also revealed significant difference in positive rate between ELISA and IHA (chi(2)=24.74, P<0.005), as well as between ELISA and COPT (chi(2)=58.14, P<0.005). These results suggest that the rSj14-3-3 and r26kDa SjGST would be effective diagnostic antigens for detection of antibodies to S. japonicum in human. Due to the easy production, high sensitivity and specificity, the recombinant proteins tested in this study can be considered as candidate reagent for immunological diagnosis of human schistosomiasis.
Luo
Qing-Li
QL
Key Laboratory of Gene Resource Utilization for Severe Diseases, Anhui Medical University, Ministry of Education, Hefei 230032, PR China.
Qiao
Zeng-Pei
ZP
Zhou
Yin-Di
YD
Li
Xiao-Yue
XY
Zhong
Zheng-Rong
ZR
Yu
Yi-Jing
YJ
Zhang
Shi-Hai
SH
Liu
Miao
M
Zheng
Mei-Juan
MJ
Bian
Mao-Hong
MH
Shen
Ji-Long
JL
D048948:14-3-3 Proteins / Q000235:genetics; D000818:Animals; D002681:China; D002846:Chromatography, Affinity / Q000379:methods; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012554:Schistosomiasis japonica / Q000175:diagnosis* / Q000453:epidemiology; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies
D048948:14-3-3 Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase