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Objective To compare and study the value of multiple antigens dot immunogold filtration assay (DIGFA ) and ima-ging diagnosis for rapid diagnosis of two kinds of echinococcosises .Methods 167 cases of hydatid patients diagnosied by pathologi-cal examination were divided into the DIGFA group for diagnosis of DIGFA and the control group for imaging diagnosis .Results The diagnosis rate of cystic echinococcosis (CE) in the DIGFA group was 74 .60% and control group was 90 .48% (P<0 .01);the diagnosis of alveolar echinococcosis(AE) in the DIGFA group was 92 .68% and the control group was 73 .17% (P<0 .05);when the cystica<5 cm ,the diagnosis rate of AE and CE in the DIGFA group was 91 .67% and 61 .11% (P<0 .05) ,when the cystica 5- <10 cm ,the detection rate of AE and CE in the DIGFA group was 94 .12% and 71 .43% (P<0 .05) .When the cystica≥10 cm ,<5 cm or between 5 - < 10 cm ,the detection rate of CE in DIGFA group was 94 .12% ,61 .11% ,71 .43 ,respectively (P<0 .05);The totle detection rates of the AE and CE in DIGFA group were 92 .68% and 74 .60% (P<0 .05) .Conclusion Imaging di-agnosis for the CE was higher and the DIGFA diagnosis for the AE was higher and the DIGFA also had clinical significance espe-cially applicated to the early diagnosis of AE .With the help of the imaging diagnosis ,the DIGFA could diagnose two kinds of echi-nococcosises correctly and it provided the benefits of specificity and sensitivity and performed easily .
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Objective To develop a rapid kit applied to the field for detection of antibody to schistosome in human sera. Methods A new kit for rapid detection of antibody to schistosome was developed through improving the dot immunogold filtration assay (DIGFA). A total of 100 cases of sera from chronic schistosomiasis patients and 140 from healthy people, HBV patients and the people infected with other parasites were detected by the kit. The sensitivity, specificity, Youden's index and Kappa value were utilized as the evaluation standard. Results The sensitivity of detecting antibody to schistosome, specificity, Youden's index and Kappa value were 92% , 95.08% , 0.87 and 0.87, respectively. The cross reaction to patients with clonorchiasis was 5%. Conclusion DICFA kit is practical for antibody to schistosome detection in the field because of its advantages such as smaller serum needed and faster in reaction.
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Objective To establish a new,rapid,simple and reliable assay for detecting autoantibody SSB.Methods A new dot immunogold filtration assay(DIGFA) was developed,in which the recombinant SSB protein expressed in Pichia pastoris was bound to nitrocellulose(NC) membrane and colloidal gold-labeled staphylococus protein A(SPA) was used as an indicator.Results The sensitivity and specificity of DIGFA were 100% and 98.75%,respectively.The agreement between DIGFA and ENA dot assay was 99.01%.Conclusion DIGFA for detecting autoantibody SSB is a good,rapid,simple and accurate assay for clinical diagnosis.
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Objective To study a new and rapid method for detection of anti-histoplasma antibody by dot immunogold filtration assay (DIGFA). Methods DIGFA was developed by coating purified protein derivative of histoplasmin (P-HTPM) on nitrocellulose membrane as membrane antigen and labeling SPA with colloidal gold. Anti-histoplasma antibodies in sera from mice immunized with Histoplasma were detected by DIGFA. Results All immunizedsera with Histoplasma were positive by DIGFA as well as the results detected by ELISA. The sera immunized respectively with Monilia and Penicillium marneffei were negtive by DIGFA while 1 of 8 immunizedsera with Blastomyces dermatitidis and 2 of 9 immunizedsera with Paracoccidioides brasiliensis were found to be positive. Conclusion DIGFA was a valuable and rapid method to detect histoplasmaantibody in serum.
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Objective To obtain recombinant human Smith D1 (Sm D1) antigen and establish detecting assay.Methods Human Smith D1 antigen was synthesized by PCR using human Leukemic cDNA. The prokaryotic expression vector pGEX-ST-Sm D1 was constructed and transformed into E.coli.BL21 cell.Protein expressed under the induction of IPTG.We established DIGFA for detecting anti-Sm D1 antibodies with purified Sm D1 antigens.Results Sequence and restriction analysis revealed Sm D1 gene was cloned in frame into pGEX-5T,SDS-PAGE profile showed a clear protein band with a relative molecular weight of 39 000 and western blotting indicated that the expressed product specifically reacted to polyclonal anti-human Sm D1 genes.There was no significant difference between DIGFA and IB.The agreement between DIGFA and IB was 91.7% as calculated by Kappa statistical method.The sensitivity and specificity of DIGFA were 100% and 83.3% repectively.Conclusions Human Sm D1 gene is successfully cloned、 expressed and purification.The DIGFA,using purified Sm D1 antigens,is as good as IB,rather simpler, more rapid and reliable assay.
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0^05). The negative rate of DIGFA in healthy people was 100%(40/40). The cross reaction rate in 20 cysticercosis cases and 25 schistosomiasis cases were 5%(1/20) and 4%(1/25), respectively. Both coincidence rates comparing DIGFA with dot\|ELISA were 90^9%(50/55). Conclusion DIGFA is as sensitive and specific as the dot\|ELISA,and has the advantages of simplicity and without specific equipment.