1.Research on electrochemical luminescence analyzer test anemia indicators of performance verification methodology
Wei YAO ; Aisheng LIU ; Duzhi FANG
China Medical Equipment 2016;13(1):74-77
Objective:To detect anaemia parameter methodology performance for validation of Roche Cobas E601 automatic electrochemical luminescence immunity analyzer.Methods:Recommended by the American association of clinical laboratory standardization (CLSI) method was developed for the determination of folic acid, iron, protein, and this precision, accuracy, linear range, sensitivity, biological reference range and carry pollution index, and validated.Results: Cohas E601 determination of folic acid, iron, protein and precision, the daytime in this batch variation coefficient were 3.03%~4.27% and 3.51%~4.68%. Relative bias must lean on(%) between -3.54%~4.46%. The scope of determination of linear range and the manufacturer to provide similar. Folic acid, iron, protein and numerical value with the determination of this instrument manufacturers provide reference interval coincidence rate were 90.0%, 85.0% and 90.0% respectivel. Instrument to detect carry pollution rate is 0.04%~0.16%. CohasE601 detection sensitivity were 0.23 ng/ml, 0.21 ng/ml and 0.19 pg/ml.Conclusion: Cobas E601 detect folic acid, iron, protein and good performance of this methodology, but manufacturers provide biological reference range is not suitable for the local crowd, should establish the corresponding normal reference range.
2.Discussion of Effect on Nitrite Test Results of Reagent Strips Placed Time on COMBI-SCAN XL Automatic Urine Analyzer
Duzhi FANG ; Aisheng LIU ; Wei YAO
Journal of Modern Laboratory Medicine 2015;(2):126-127,131
Objective To explore the division XL automatic urine analyzer reagent strips placed time impact on nitrite (NIT) test results,to ensure that provide accurate and reliable experimental data for clinical,and improve clinical diagnostic rate. Methods Used a new load reagent storehouse article in the new reagents respectively in <5,30 min;1,2,3,3.5,4,4.5,5, 5.5,6,6.5,7 and 8 h,in different period,16 0.9 g/dl saline NIT in the false positive rate,and carried on the comparison to different times NIT false-positive rate analysis.Results <3.5 h NIT false positive rate was 0% (0/16),4~4.5 h false pos-itive rate was 18.8% (3/16),5~5.5 h false positive rate was 43.8% (7/16),6~6.5 h was 62.5% (10/16),7~8 h was 87.5% (14/16).More than 4 h each time comparison between NIT false positive rate,differences were statistically signifi-cant (χ2 =11.7~59.2,all P <0.01).Conclusion Division XL automatic urine analyzer reagent strips placed after 4 h,NIT false-positive rate started to rise,NIT false-positive rate was significantly increased after 5 h.Therefore,strengthen the divi-sion XL automatic urine analyzer reagent strips placed reasonable time management,should be in the < 3.5 h advisable,it helps reduce the NIT false positive rate.
3.Present Situation Investigation of Red Blood Cells Glucose-6-Phosphate Dehydrogenase Dificiency of Pregnancy Couple in Baoan District of Shenzhen
Aisheng LIU ; Duzhi FANG ; Wei YAO
Journal of Modern Laboratory Medicine 2015;(3):157-158,161
Objective To understand red blood cells glucose-6-phosphate dehydrogenase (G6PD)dificiency of pregnancy cou-ple in baoan district of Shenzhen,in order to draw the attention of society.Methods Collected 6 574 cases of couples of child-bearing age (3 192 males and 3 382 females),in Baoan District People’s Hospital,from February to November 2013. Venous blood was collected using EDTA anticoagulant blending respectively,the improvement of red bloodcells and glucose-6-phosphate dehydrogenase quantitative ratio method to determine anticoagulant G6PD/6 PGD ratio in the whole blood,with ratio <1.0 convicted of G6PD deficiency.Results 3 192 cases of male subjects,G6PD deficient 176 cases,3 382 cases of pregnant women client detection of 135 cases in the detection rate were 5.51% and 3.99%,respectively,the total detection rate was 4.73% (311/6 574),G6PD lack of rate was higher than that of pregnant women,men G6PD lack of male∶female was 1.30∶1 (176∶135),the difference was statistically significant between (χ2 =7.16,P <0.05).Conclusion The baoan district couple G6PD lack had a higher incidence of pregnancy,men were significantly higher than women,should arouse at-tention.Couples pregnancy test during pregnancy should be advocated the G6PD activity screening,the eugenics and mater-nal and child health care has important significance.
4.Cloning Recombinant Expression of Treponema Pallidum Four Kinds of Membrane Protein and Application of Establishing ELISA Method
Duzhi FANG ; Jian YANG ; Rongzhi YANG ; Cheng TANG
Journal of Modern Laboratory Medicine 2015;(1):75-77,81
Objective To clone Tp0821,Tp0319,Tp0971 and Tp0663 gene,construct prokaryotic expression plasmid,the ex-pression of recombinant proteins,purification and used to detect syphilis patients serum antibodies.To explore Tp0821, Tp0971,Tp0319 and Tp0663 recombinant protein in the diagnosis of Tp infection,rich library of candidate antigens syphilis serology diagnosis.Methods From Tp library screening Tp0821,Tp0319,Tp0971 and Tp0663 four kinds of membrane pro-tein,through the analysis of the bioinformatics software selection advantage antigen epitope,connected with pQE32 carrier respectively built pQE32/Tp0821,pQE32/Tp0319,pQE32/Tp0971 and pQE32/Tp0663 prokaryotic expression recombi-nant;In vitro cloning,expression and purification and on single Tp0821,Tp0319,Tp0971,Tp0663 and four kinds of recombi-nant protein chimeric antigen (Tp0821-Tp0319-Tp0971-Tp0663)in order to establish the indirect Tp-envelope antigen ELISA method,based on TRUST and TPPA method comparison,detection collected from January 2013 to June 2014 in Shenzhen Baoan District People’s Hospital outpatient and hospitalization Tp negative patients,160 cases of positive speci-mens of 83 cases of clinical and Tp,and evaluated its application in the syphilis serology diagnosis.Results Successful con-structed of prokaryotic expression vector pQE32/Tp0821,pQE32/Tp0319,pQE32/Tp0971 and pQE32/Tp0663.Efficient expressed and purified their relative molecular mass of recombinant proteins.Seted up Tp-indirect ELISA method to detect 160 cases of Tp negativepositive clinical specimens and 83 cases of Tp,and compared with TPPA,the sensitivity were 85.5% (71/83),84.3% (70/83),89.2% (74/83),81.9% (68/83)and 95.2% (79/83)respectively,specificity was 100%(160/160),and the coincidence rate was 82.6%~95.7%.Single Tp0821,Tp0319,Tp0971 and Tp0663 recombinant protein positive rate of TP-ELISA was established (85.5%,84.3%,89.2% and 81.9%)compared with TPPA method of positive detection rate (100%)had significant difference statistically significant (χ2= 24.5~31.8,P<0.01),and four kinds of re-combinant protein chimeric antigen positive rate of TP-ELISA was established with the TPPA method the positive detection rate of comparative difference was statistically significant (χ2=7.95,P<0.05).Conclusion Preparation Tp0821,Tp0319, Tp0971 and Tp0663 recombinant protein had good immune activity,for the further study of its application in the diagnosis of syphilis serology lay a certain foundation,but in four kinds of recombinant protein chimeric antigen based Tp Tp-indirect ELISA method for serological diagnosis,can improve the detection sensitivity.