目的探讨并验证EURO Blot Master Ⅱ(简称EBM II)全自动免疫印迹仪与AUTOBLOT SYSTEM 20(简称AS20)蛋白印迹仪同时进行HIV初筛阳性标本确证实验结果的一致性与相关性,评估EBM Ⅱ全自动免疫印迹仪可以满足HIV抗体确证实验的要求,可以在...目的探讨并验证EURO Blot Master Ⅱ(简称EBM II)全自动免疫印迹仪与AUTOBLOT SYSTEM 20(简称AS20)蛋白印迹仪同时进行HIV初筛阳性标本确证实验结果的一致性与相关性,评估EBM Ⅱ全自动免疫印迹仪可以满足HIV抗体确证实验的要求,可以在中国各级疾病预防控制中心及通过认证的HIV确证实验室中使用。方法采用MP公司AS20蛋白印迹仪作为参比仪器,欧蒙EBM Ⅱ全自动免疫印迹仪为实验仪器,两台仪器同时对71例HIV初筛阳性血清标本进行检测。计算其阳、阴性符合率,验证两种仪器对同一样本检测结果的一致性与相关性。结果两台仪器的阳、阴性符合率均达到100%,阳、阴性预测值也都达到了100%。结论 EBM II与AS20的检测结果具有很好的一致性及相关性,完全符合各级疾病预防控制中心及HIV确证实验室的用于HIV抗体确证实验检测的使用要求。展开更多
HIV-indeterminate Western blotting(WB)results are typically obtained in WB confirmatory assays,and the number of indeterminate samples may increase with the detection of HIV infections,which will present considerable ...HIV-indeterminate Western blotting(WB)results are typically obtained in WB confirmatory assays,and the number of indeterminate samples may increase with the detection of HIV infections,which will present considerable challenges for the management of HIV/AIDS.Nucleic acid detection has been used as a laboratory test for screening suspected or indeterminate samples.However,the effectiveness of these assays for the differential diagnosis of HIV-indeterminate WB samples remained undetermined.In this study,210 subjects with HIV-indeterminate WB results were detected from 6360 positive HIV screening samples between 2015 and 2016 in southeastern China,in which HIV-indeterminate WB results accounted for 3.30%.The highest proportion of indeterminate results was observed in pregnant and lying-in women receiving physical examinations(16.67%),followed by that in voluntary blood donors(8.82%).The most common WB band patterns were p24,gpl60 and p24,and gpl60.The follow-up study revealed that the highest negative and positive conversion rates of HIV antibodies were in samples with a single p24 band(80.28%),and with gpl60 and p24 bands(86.21%),respectively.Among the Env,Gag,and Pol antibodies,samples with a Gag band showed the highest negative conversion rate(81.25%),whereas the highest positive conversion rate was observed in samples with an Env band(56.76%).In addition,quantitative and qualitative HIV nucleic acid testing exhibited the highest sensitivity(96.3%)and specificity(97.85%),respectively.Our results indicate a lower proportion of HIV indeterminate WB results in southeastern China compared to previous reports,and the follow-up re-examination of patients with HIV indeterminate results should be performed.Nucleic acid testing facilitates the identification of HIV infections.展开更多
Objective Western blotting （WB;immunoblotting） is a widely used tool for the serodiagnosis of Lyme borreliosis （LB）,but so far,no generally accepted criteria for its performance and interpretation have been establ...Objective Western blotting （WB;immunoblotting） is a widely used tool for the serodiagnosis of Lyme borreliosis （LB）,but so far,no generally accepted criteria for its performance and interpretation have been established in China.The present study was designed to determine the criteria for standardized Western blot for the predominant species of Borrelia burgdorferi sensu lato in China,in which WB was produced with strain PD91 as the representative strain attributed to predominant genospecies Borrelia garinii of Borrelia burgdorferi sensu lato.Methods Approximately 13 bands between 14 and 100 kD were differentiated for strain PD91 by using Gel-Pro analysis software.In a study with 631 serum samples （taken from 127 patients with Lyme borreliosis and 504 controls）,all observed bands were documented.To establish criteria for a positive WB result for strain PD91,receiver operating characteristic （ROC） curves were used.Results The following interpretation criteria were recommended：for IgG,at least one band of P83/100,P58,P39,P30,OspC,P17,P66,and OspA;for IgM,at least one band of P83/100,P58,OspA,P30,OspC,P17 or P41.In addition,syphilis,leptospirosis and other related diseases should be excluded when the positive band is P41 in IgM.For IgG criteria,the sensitivity is 73.2%,the specificity is 99.4% and Youden index is 0.726;for IgM criteria,the sensitivity is 50.6%,the specificity is 93.1% and Youden index is 0.437.Conclusion Standardization of WB assays is necessary for comparison of results from different laboratories.Moreover,the criteria of other genospecies of Borrelia burgdorferi sensu lato should be determined in the future to complete the criteria of WB for the diagnosis of the Lyme disease in China.展开更多
Objective To study the technique of Western blot for the diagnosis of Lyme disease caused by Borrelia afzelii in China and to establish the standard criteria by operational procedure. Methods FP1, which is the represe...Objective To study the technique of Western blot for the diagnosis of Lyme disease caused by Borrelia afzelii in China and to establish the standard criteria by operational procedure. Methods FP1, which is the representative strain of B. afzelii in China, was analyzed by SDS-PAGE, electro transfer and irnmunoblotting assays. The molecular weights of the protein bands of FP1 were analyzed by Gel-Pro analysis software. In a study using 451 serum samples （159 patients with Lyme disease and 292 controls）, all observed bands were recorded. The accuracy of the WB as a diagnostic test was established by using the ROC curve and Youden index. Results Criteria for a positive diagnosis of Lyme disease were established as at least one band of P83/100, P58, P39, OspB, OspA, P30, P28, OspC, P17, and P14 in the IgG test and at least one band of P83/100, P58, P39, OspA, P30, P28, OspC, P17, and P41 in the IgM test. For IgG criteria, the sensitivity, specificity and Youden index were 69.8%, 98.3%, and 0.681, respectively; for IgM criteria, the sensitivity, specificity and Youden index were 47%, 94.2%, and 0.412, respectively.展开更多
Objective To establish and optimize the proteomic analysis of protoscoleces-specific antigens from Echinococcus granulosus. To provide a foundation for identifying specific antigens in the soluble proteins of E. granu...Objective To establish and optimize the proteomic analysis of protoscoleces-specific antigens from Echinococcus granulosus. To provide a foundation for identifying specific antigens in the soluble proteins of E. granulosus protoscoleces for further research. Methods Brood capsules were collected aseptically from fertile E. granulosus cysts from the livers of an infected patient. The fertile E. granulosus cysts were fractured, and protoscoleces were collected by centrifugation. The soluble proteins of protoscoleces were acquired using the 2D Quant kit according to the manufacturer＇s instructions. We employed two-dimensional electrophoresis （2-DE） combined with immunoblot assay （Western blot） to analyze the soluble components of E. granulosus protoscoleces antigens. The 2-DE and immunoblot maps obtained were analyzed with PDQuest 8.0 image analysis software. Results About 233 soluble protein spots were identified with Coomassie-stained gels. Most of the proteins had a molecular weight of 16 000 Da to 117 000 Da, and an isoelectric point value of 3.0 to 10.0. 2-DE immunoblot was conducted and 57 specific antigen spots were observed, among which 23 spots were identified. Conclusion 2-DE combined with Western blot is the key to successful proteomic analysis and presents a new possibility for searching the specific E. granulosus protoscoleces antigens.展开更多
基金supported by grants from the Cultivation of Young Talents Project Fund from the Fujian Provincial Health and Family Planning Commission (Grant No. 2015-ZQN-ZD11)the Pilot Project of Fujian Provincial Department of Science and Technology (Grant No. 2016Y0010)+2 种基金the Jiangsu Provincial Project of Invigorating Health Care through Science, Technology and EducationJiangsu Provincial Medical Youth Talentthe Project of Invigorating Health Care through Science, Technology and Education (Grant No. QNRC2016621)
文摘HIV-indeterminate Western blotting(WB)results are typically obtained in WB confirmatory assays,and the number of indeterminate samples may increase with the detection of HIV infections,which will present considerable challenges for the management of HIV/AIDS.Nucleic acid detection has been used as a laboratory test for screening suspected or indeterminate samples.However,the effectiveness of these assays for the differential diagnosis of HIV-indeterminate WB samples remained undetermined.In this study,210 subjects with HIV-indeterminate WB results were detected from 6360 positive HIV screening samples between 2015 and 2016 in southeastern China,in which HIV-indeterminate WB results accounted for 3.30%.The highest proportion of indeterminate results was observed in pregnant and lying-in women receiving physical examinations(16.67%),followed by that in voluntary blood donors(8.82%).The most common WB band patterns were p24,gpl60 and p24,and gpl60.The follow-up study revealed that the highest negative and positive conversion rates of HIV antibodies were in samples with a single p24 band(80.28%),and with gpl60 and p24 bands(86.21%),respectively.Among the Env,Gag,and Pol antibodies,samples with a Gag band showed the highest negative conversion rate(81.25%),whereas the highest positive conversion rate was observed in samples with an Env band(56.76%).In addition,quantitative and qualitative HIV nucleic acid testing exhibited the highest sensitivity(96.3%)and specificity(97.85%),respectively.Our results indicate a lower proportion of HIV indeterminate WB results in southeastern China compared to previous reports,and the follow-up re-examination of patients with HIV indeterminate results should be performed.Nucleic acid testing facilitates the identification of HIV infections.
文摘目的对Western Blot和流式细胞术(Flowcytometry,FCM)这两种磷酸化STAT3的检测方法进行比较。方法用不同浓度的白细胞介素5(interleukin 5,IL-5)刺激人早幼粒细胞白血病细胞株HL-60,活化细胞内的STAT3,然后分别用Western Blot和流式细胞术进行检测。结果 Western Blot与流式细胞术的检测结果一致。结论 Western Blot与流式细胞术用于磷酸化STAT3的检测各有优势,可以互为补充。
基金supported by the "Tenth Five-Year Plan" project of research on the specific diagnosis method for the Lyme disease. 2001BA705B07
文摘Objective Western blotting （WB;immunoblotting） is a widely used tool for the serodiagnosis of Lyme borreliosis （LB）,but so far,no generally accepted criteria for its performance and interpretation have been established in China.The present study was designed to determine the criteria for standardized Western blot for the predominant species of Borrelia burgdorferi sensu lato in China,in which WB was produced with strain PD91 as the representative strain attributed to predominant genospecies Borrelia garinii of Borrelia burgdorferi sensu lato.Methods Approximately 13 bands between 14 and 100 kD were differentiated for strain PD91 by using Gel-Pro analysis software.In a study with 631 serum samples （taken from 127 patients with Lyme borreliosis and 504 controls）,all observed bands were documented.To establish criteria for a positive WB result for strain PD91,receiver operating characteristic （ROC） curves were used.Results The following interpretation criteria were recommended：for IgG,at least one band of P83/100,P58,P39,P30,OspC,P17,P66,and OspA;for IgM,at least one band of P83/100,P58,OspA,P30,OspC,P17 or P41.In addition,syphilis,leptospirosis and other related diseases should be excluded when the positive band is P41 in IgM.For IgG criteria,the sensitivity is 73.2%,the specificity is 99.4% and Youden index is 0.726;for IgM criteria,the sensitivity is 50.6%,the specificity is 93.1% and Youden index is 0.437.Conclusion Standardization of WB assays is necessary for comparison of results from different laboratories.Moreover,the criteria of other genospecies of Borrelia burgdorferi sensu lato should be determined in the future to complete the criteria of WB for the diagnosis of the Lyme disease in China.
基金supported by the 12th Five-Year Major National Science and Technology Projects of China (No.2011ZX10004-001)Natural Science Foundation of China (31100105)
文摘Objective To study the technique of Western blot for the diagnosis of Lyme disease caused by Borrelia afzelii in China and to establish the standard criteria by operational procedure. Methods FP1, which is the representative strain of B. afzelii in China, was analyzed by SDS-PAGE, electro transfer and irnmunoblotting assays. The molecular weights of the protein bands of FP1 were analyzed by Gel-Pro analysis software. In a study using 451 serum samples （159 patients with Lyme disease and 292 controls）, all observed bands were recorded. The accuracy of the WB as a diagnostic test was established by using the ROC curve and Youden index. Results Criteria for a positive diagnosis of Lyme disease were established as at least one band of P83/100, P58, P39, OspB, OspA, P30, P28, OspC, P17, and P14 in the IgG test and at least one band of P83/100, P58, P39, OspA, P30, P28, OspC, P17, and P41 in the IgM test. For IgG criteria, the sensitivity, specificity and Youden index were 69.8%, 98.3%, and 0.681, respectively; for IgM criteria, the sensitivity, specificity and Youden index were 47%, 94.2%, and 0.412, respectively.
基金supported by the National Natural Science Foundation of China (Nos. 30260105 and 30660176)the Natural Science Foundation of Ningxia Hui Autonomous Region (NZ10192)the Doctor’s Subject Science Foundation of Ningxia Medical University (KF2010-33)
文摘Objective To establish and optimize the proteomic analysis of protoscoleces-specific antigens from Echinococcus granulosus. To provide a foundation for identifying specific antigens in the soluble proteins of E. granulosus protoscoleces for further research. Methods Brood capsules were collected aseptically from fertile E. granulosus cysts from the livers of an infected patient. The fertile E. granulosus cysts were fractured, and protoscoleces were collected by centrifugation. The soluble proteins of protoscoleces were acquired using the 2D Quant kit according to the manufacturer＇s instructions. We employed two-dimensional electrophoresis （2-DE） combined with immunoblot assay （Western blot） to analyze the soluble components of E. granulosus protoscoleces antigens. The 2-DE and immunoblot maps obtained were analyzed with PDQuest 8.0 image analysis software. Results About 233 soluble protein spots were identified with Coomassie-stained gels. Most of the proteins had a molecular weight of 16 000 Da to 117 000 Da, and an isoelectric point value of 3.0 to 10.0. 2-DE immunoblot was conducted and 57 specific antigen spots were observed, among which 23 spots were identified. Conclusion 2-DE combined with Western blot is the key to successful proteomic analysis and presents a new possibility for searching the specific E. granulosus protoscoleces antigens.
