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Objective:To establish reverse triiodothyronine (rT 3) biological reference interval suitable for laboratory by indirect method. Methods:From April to September 2019, 797 cases (332 males, 465 females, age: 12-95 years) underwent thyroid function, thyroid related antibody and rT 3 tests from hospitalized population in Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine were retrospectively analyzed. The reference individuals with normal thyroid hormone, antibody and without thyroid nodule or goiter were screened as inclusion criteria, and the factors such as acute and chronic diseases or drugs that might affect the values of rT 3 were excluded. Independent sample t test, one-way analysis of variance and least significant difference t test were used to analyze data. The rT 3 reference interval was established by non-parametric sequencing method, and 2.5% and 97.5% percentile values of data distribution were selected as the upper and the lower reference limits. In order to verify the rT 3 reference interval, 20 healthy individuals and 20 inpatients who met the inclusion and exclusion criteria were selected to test rT 3 with a simple random sampling method. Results:A total of 159 reference individuals (66 males, 93 females, age: 23-87 years) were enrolled. The rT 3 values of 23-29( n=4), 30-39( n=18), 40-49( n=29), 50-59( n=43), 60-69( n=40), 70-79( n=19) and over 80( n=6) years old groups were (0.62±0.16), (0.63±0.12), (0.64±0.11), (0.61±0.11), (0.65±0.14), (0.65±0.11) and (0.79±0.10) μg/L, respectively. There was a statistically significant difference in the rT 3 test results among different age groups ( F=2.17, P=0.049). There were statistically significant differences of rT 3 between the individuals over 80 years old and other age groups (all P<0.05), while there were no statistically significant differences among the other groups (all P>0.05). The rT 3 of males and females under 80 years old were (0.62±0.11) and (0.64±0.12) μg/L, respectively, with no significant difference between them ( t=-0.81, P=0.420). The newly established rT 3 reference interval suitable for people above 20 years old and below 80 years old was 0.47-0.92 μg/L, and the lower limit was significantly higher than that of the reference interval in the reagent specification (0.20-0.95 μg/L). The rT 3 range of 20 healthy individuals was 0.57-0.82 μg/L and that of 20 inpatients was 0.48-0.77 μg/L, which were all within the new reference interval. Conclusion:The rT 3 biological reference interval established here has clinical application value, but its applicable range of age still needs to be further improved.
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Objective:To establish a quantitative detection method for the main components of dust mite allergens Der p 1, Der p 2 specific immunoglobulin E (sIgE) by using the nano-magnetic particle chemiluminescence immunoassay.Methods:The performance indexes of the established method were evaluated after setting up and optimizing the chemiluminescence detection system and immune reaction conditions of sIgE for dust mite allergen. Serum sIgE levels of 50 suspected allergic patients with dust mite were determined by this chemiluminescence method. At the same time, this method was compared with the Phadia kit and the consistency was analyzed by Kappa test. Results:The optimal amount of magnetic beads was 25 μg, the optimal reaction buffer (pH=7.4) contained 0.1 mol/L Tris-HCl and 0.25%( W/ W) casein, the optimal coating solution contatined 20 mmol/L phosphate buffer (PB) and 1%( W/ W) bovine serum albumin (BSA), and the luminescence enhancement solution contained 0.05%( V/ V) Triton X-100. The two-step immunoreaction was adopted, and the detection could be completed with 20 μl sample at the optimal reaction temperature of 37℃. The limit of detection (LOD) of the established nano-magnetic particle chemiluminescence system in detecting Der p 1 and Der p 2 sIgE antibodies were both less than 0.01 kU/L, with the linear range of 0.2-100.0 kU/L, the precision of less than 7%, and the cross contamination rate of 0.19% and 0.21%. Compared with the Phadia system, the positive and negative coincidence rate of Der p 1 were 78.0%(32/41) and 9/9 with good consistency ( Kappa=0.65, P=0.008), and the positive and negative coincidence rate of Der P 2 were 93.3%(28/30) and 85.0%(17/20) with good consistency ( Kappa=0.79, P=0.003). Conclusion:The nano-magnetic particle chemiluminescence immunoassay is successfully established for detecting dust mite allergen sIgE, which has good detection performance and good consistency with Phadia system.
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Objective:To evaluate the measurement agreement of Roche 25(OH)D immunoassay(evaluation method) with LC-MS/MS (reference method).Methods:A total of 909 residual serum samples from routine health check participants were collected from May to June in 2019. 25(OH)D concentrations were measured by evaluation method and LC-MS/MS, respectively. Passing-bablok regression, intraclass correlation coefficient (ICC), Bland Altman plots and Kappa test were used to analyze the consistency and bias on the results derived from the two measurement methods.Results:The 25(OH)D concentration derived from evaluation method was significantly different from those from LC-MS/MS method ( P<0.001). Slope of regression for evaluation method and LC-MS/MS was 0.962(95% CI 0.919-1.007), while intercept was -0.185 (95% CI -1.191-0.745). The ICC was 0.765 (95% CI 0.735-0.792). Altman plot showed that the average deviation between evaluation method and LC-MS/MS was -0.902 ng/ml (0.300%). The coincidence rate of evaluation method′s judgment of vitamin D sufficiency, insufficiency and deficiency with LC-MS/MS was 83.39%, and the weighted Kappa values was 0.790. Conclusion:Roche automatic 25(OH)D immunoassay shows acceptable correlation and agreement with LC-MS/MS, however, it is to note that the deviation between immunoassay and LC-MS/MS may lead to wrong judgment of vitamin D nutritional status. It is recommended that each laboratory should establish own corresponding reference values for 25(OH)D concentrations derived from these two methods.
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Anti-Müllerian hormone (AMH),a dimer glycoprotein secreted by sustentacular cells of testis and ovarian granulosa cells,belonging to the transforming growth factor β super-family.AMH is able to regulate follicular development and participate in follicular growth process,and it is relatively constant throughout the menstrual cycle compared with other ovarian reserve indicators.At present,AMH is widely used to evaluate ovarian reserve and to diagnose and evaluate the development and prognosis of ovarian diseases.It has been increasingly applied in the field of female assisted reproduction in recent years.With the development of detection technology,the sensitivity and accuracy of methods for detecting AMH are gradually improved.This review summarizes the research background,mechanism of action,clinical applications and detecting methods of AMH.
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Objective To study the changes and characteristics of TPO -Ab and TG-Ab in type 2 diaetic patients and provide new ideas for the diagnosis of diabetes.Methods From January 2014 to January 2017,160 samples in General Hospital of Taiyuan Iron&Steel (Group) Co.Ltd were selected,80 healthy people and 80 patients with type 2 diabetes,fasting venous 5 mL blood was obtained in the morning ,then electrochemical luminescence method was used to test TPO-Ab and TG-Ab contents.The diabetic patients were divided into four groups :TPO-Ab normal group,TPO-Ab elevation group,TG-Ab normal group,TG-Ab elevation group.The blood glucose,age and gender of the four groups were compared.Results Compared with the control group ,the proportions of increased TPO -Ab and TG-Ab in diabetic patients were 11.25%and 2.5%respectively,the difference was statistically significant (χ2＝4.86,P＜0.05).In type 2 diabetic patients,the blood glucose value of the normal TPO -Ab group was (6.67 ± 1.53)mmol/L,which in the TPO -Ab elevation group was (7.87 ±1.24) mmol/L,the difference was statistically significant (t＝2.94,P＜0.05).The blood glucose of the normal TG -Ab group was (6.75 ±1.34)mmol/L,which in the TG-Ab elevation group was (7.04 ±1.25)mmol/L,the difference was statistically significant (t＝2.82,P＜0.05).TPO-Ab and TG-Ab elevation had no obvious relation with age ,gender.The age of the normal TPO -Ab group was (62.1 ±6.3)years,which in the TPO-Ab elevation group was (63.0 ±4.9)years,there was no statisti- cally significant difference (t＝1.37,P＞0.05).The age of the normal TG -Ab group was (62.8 ±7.1)years,which in the TG-Ab elevation group was (61.6 ±2.7)years,the difference was not statistically significant (t＝1.27,P＞0.05).In male patients,TPO-Ab normal accounted for 84.09%,TPO-Ab rise accounted for 15.91%.In female patients,TPO-Ab normal accounted for 86.11%,TPO-Ab rise accounted for 13.89%,there was no statistically significant difference (χ2＝1.20,P＞0.05).In male patients,TG-Ab normal accounted for 97.73%,TG-Ab rise accounted for 2.27%, in female patients, TG -Ab normal accounted for 97.22%, TG -Ab rise accounted for 2.78%,there was no statistically significant difference (χ2＝0.97,P＞0.05).Conclusion TPO-Ab and TG-Ab in type 2 diabetes patients are higher than healthy people.The increase of TPO -Ab and TG -Ab is positively correlated with blood glucose level.The increase of TPO-Ab and TG-Ab is not correlated with age and gender.
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Objective To validate the performance of 4 domestic chemiluminescence immunoassay (CLIA) systems on 8 tumor markers quantitative assay kits. Methods Four domestic CLIA systems were randomly marked as A, B, C, D and 8 tumor markers, including carbohydrate antigen (CA)125, CA15-3, CA19-9, ferritin (Fer), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), prostate-specific an-tigen (PSA) and free PSA (fPSA) were determined. According to the standard of Clinical and Laboratory Standards Institute (CLSI), the precision, methodological comparison and analytical measure range of 4 systems were validated. Clinical serum samples were obtained from patients in Suzhou Hospital. According to the CLSI EP9-A3 protocol, imported equipment was used as the reference system. The biases of medical de-cision points were assumed, and Pearson correlation analysis and Spearman correlation analysis were used to analyze the data. Results The precision verification of CA125 and PSA on A, CA125 and AFP on B, CA125, CEA, AFP and PSA on C, and all 8 tumor markers on D could meet the laboratory quality control requirements. The correlations of the test results between A-D and the imported equipment were significant (all P<0.05) with the correlation coefficients 0.79-0.99, 0.47-0.99, 0.90-0.98 and 0.78-1.00, respec-tively, and the number of acceptable tests at the level of medical decision was 5, 2, 5, 4. All tests were certified to meet the analytical measure range validation. Conclusions The detection performance of 4 do-mestic CLIA systems for all 8 tumor markers are different. The performance of domestic CLIA systems should be tested when choosing one that can meet laboratory quality control requirements.
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Objective To study the comparability of total prostate specific antigen ( tPSA) meas-urement by four domestic chemiluminescence immunoassays ( DCI) and electrochemiluminescence immuno-assay ( ECI) . Methods A total of 45 serum samples that requested tPSA tests were selected. Four DCIs ( Snibe MAGLUMI 4000, Mindray CL-2000i, Autobio A2000, HYBIOME AE180) and ECI ( Roche Cobas e601) were used to measure tPSA. The precisions of the methods were evaluated. The four DCIs were com-pared with Roche ECI respectively, and the comparability of the test results was analyzed. Wilcoxon signed rank test and Spearman correlation analysis were used to analyze the data. Results The precisions of five methods were good. The tPSA levels measured by Roche Cobas e601, Snibe MAGLUMI 4000, Mindray CL-2000i, Autobio A2000, and HYBIOME AE180 were 14.11(9.92, 36.09), 12.00(8.56, 27.23), 12.10 (8. 60, 29.87), 13.35(9.51, 32.85) and 14.50(9.88, 40.06) μg/L, respectively. The correlation coeffi-cients of Roche with Snibe MAGLUMI 4000, Mindray CL-2000i, and Autobio A2000 were 0.992, 0.989, 0. 957 and 0.983, respectively (all P<0.001). Assuming the tPSA medical decision point for regression equation was 4.0μg/L, the proportional biases of Snibe MAGLUMI 4000, Mindray CL-2000i, Autobio A2000, and HYBIOME AE180 compared with Roche were -10. 88％, -18. 07％, 0. 23％ and 22. 31％, respectively. Conclusion The comparability of tPSA test results is different between 4 DCIs and Roche ECI, which pro-vides some references for clinical application and standardization of the DCI test results.
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Objective To establish a fast and quantitative light initiated chemiluminescent assay (LICA) method for high mobility group box1 (HMGB1) determination.Methods Two strains of paired HMGB 1 monoclonal antibodies were used.One was used to coat receptor microspheres.The other was labeled with biotin first and then composed with chain mildew element of affinity donor microsphere to form LICA method for HMGB1.After optimizing the reaction system,the technical specifications of the method was evaluated.Serum HMGB1 levels of common pneumonia patients (CPP) and severe pneumonia patients (SPP) were measured and compared with that of health controls.Two-sample t test was used.Results The sensitivity of LICA was 0.1 μg/L,with linear measurement ranging from 0.1 to 1 000 μg/L.The precisions of intra-and inter-analysis were 1.74％-2.92％ and 1.93％-3.73％ respectively,both were lower than 5％.The recovery rate was 99.74％ (range:94.53％-106.37％).The correlation coefficient of LICA and enzyme-linked immunosorbent assay (ELISA) was 0.888 2.The LICA method had good specificity and no obvious cross reaction with HMGB2 and HMGB3.The serum HMGB1 level in CPP (n=35) and SPP (n=25) was significantly higher than that in health controls (n=35):(6.76±3.13),(19.69±+9.04) vs (1.49±+0.74) μg/L;t values:-5.447 and-5.186,both P＜0.01.The HMGB1 levels between CPP and SPP were also significantly different (t=-3.500,P＜0.01).Conclusions The established LICA method of HMGB1 has high sensitivity and specificity with reliable results.This method is also homogeneous,fast and cleaning-free,thus has a good prospect in clinical application.
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Objective To develop a time-resolved fluorescent microspheres immunochromatographic assay (TRFMIA) for detection of alphafetoprotein (AFP) and carcinoembryonic antigen (CEA) in human serum and to evaluate its performance.Methods The Eu-time-resolved fluorescent polystyrene particles conjugated with monoclonal antibody AC18# for AFP and AE03# for CEA were used as fluorescent labels.The monoclonal antibody AC17# for AFP,AE05# for CEA and goat anti-rabbit antibody were immobilized on the nitrocellulose membrane as the test lines and control line.Several performances indicators were measured,including linear range,detection limit,and specificity.AFP and CEA were measured by the new method and the results were compared with those obtained by time-resolved fluoroimmunoassay (TRFIA) and electro-chemiluminescence immunoassay (ECLIA) using linear correlation analysis.Results The measurement ranges of AFP were 0.07-1 000.00 kU/L with the intra-and inter-assay CV of 5.93％ and 11.07％,and those of CEA were 0.12-500.O0 μg/L with the intra-and inter-assay CV of 7.53％ and 12.13％ respectively.The average recovery rate of AFP and CEA was 92.77％ and 94.73％,respectively.Measurements obtained by TRFMIA had strong correlation coefficients ranging from 0.93 to 0.97 when compared with those obtained by TRFIA and ECLIA.Conclusion TRFMIA,which can simultaneously detect AFP and CEA,has been successfully established.
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Objective To evaluate the differences of serum TSH of suspicious subclinical hypothyroidism determined by four automatic biochemical analyzers and the impact on clinical diagnosis and treatment.Methods Taking results of Roche Cobas e601 laboratory test as a reference, 103 serum samples with TSH 2.50-10.00 mU/L(90 with TSH≥4.27 mU/L) and normal FT3, FT4 were selected.Four different automatic biochemical analyzers (Cobas e601, Immulite2000, Centaur XP, I2000) were used to measure TSH of the serum samples at the same time.Wilcoxon signed rank test, Spearman correlation analysis were used for data analysis.Results TSH (M(P25, P75)) measured by 4 methods were 5.20(4.73, 6.40), 2.95(2.59, 3.48), 3.30(2.94, 4.15) and 4.10(3.43, 4.75) mU/L, which varied significantly from one assay to another (z values:-8.78,-8.41,-7.64,-8.09,-8.50, all P<0.05).The correlations between methods were of great differences (rs ranged from 0.45 to 0.92).Significant differences existed in each other for subclinical hypothyroidism diagnosis based on TSH cutoff respectively.Conclusion Results from different automatic immunoassay analyzers in patients with TSH of 2.50-10.00 mU/L varied widely, hence, it is indeterminate to diagnose subclinical hypothyroidism only relies on a single serum TSH test.
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Objective To develop a chemiluminescence imaging DNA microarray method for simultaneous,fast and accurate detection of nine rash-and fever-causing pathogens,namely,Measles virus,Rubella virus,Enterovirus type 71, Varicella zoster virus,Dengue fever virus,Human small FDNA virus B19,Coxsackie virus type A16,A-βStreptococcus pyogenes (hemolytic streptococcus)and Salmonella typhi.Methods Primers and probes were designed based on the specific sequence in the conserved region of genomes of the nine pathogens.The nucleic acids of the nine pathogens were amplified and labelled by multiplex PCR method.The multiplex PCR amplification products were hybridized with specific probes of microarray that was scanned after washing and chemiluminescence coloration to identify the nine pathogens.After the optimization of the multiplex PCR system,hybridization and chemiluminescence imaging,the specificity,sensitivity and reproducibility of the chip were evaluated.The serial diluted nucleic acid of Enterovirus type 71 was detected using microarray and real-time PCR approach to compare the sensitivity of these two methods.Results Nine specific primers and eleven specific probes were selected.The microarray demonstrated high sensitivity and specificity.The minimum detection limit of plasmid DNA and in vitro transcribed RNAs was 3 ×103 copies per reaction.The detection sensitivity of this microarray was 10 percent of that by the real-time PCR method.The rate of sensitivity and specificity of clinical sample detection was 95% and 85.7% respectively,and the rate of accuracy was 93.2%.Conclusion A chemiluminescence imaging DNA microarray method for simultaneous,fast and accurate detection of nine pathogens that cause rash and fever illnesses is established successfully,which can serve as a new high throuthput screening method for clinical diagnosis and epidemiological investigation of rash and fever illnesses.
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Objective To compare the clinical value of two kinds of imunoassay method including CLIA and ELISA in the detection of related antibody of patients with HCV.Methods 60 patients with HCV were chosen as study subjects,and the related antibodies of patients were detected by CLIA and ELISA.The positive rates of anti HCV antibody,anti-3E antibody,anti-GP210 antibody,anti-SP100 antibody,anti-PML antibody and anti-AMA -M2 antibody detected by CLIA and ELISA were compared.Results The positive rates of anti-HCV antibody detected by CLIA and ELISA were 83.33％ (50/60),98.33％ (59/60),respectively.The anti-HCV antibody detected by CLIA was significantly higher than by ELISA (x2 =4.73,P =0.03).The positive rates of anti-3E antibody,anti-GP210 antibody,anti-SP100 antibody,anti-PML antibody and anti-AMA-M2 antibody detected by ELISA were 11.67 ％ (7/60),6.67 ％ (4/60),11.67 ％ (7/60),10.00 ％ (6/60),11.67 ％ (7/60),respectively,those detected by CLIA were 73.33 ％ (44/60),50.00％ (30/60),38.33 ％ (23/60),58.33 ％ (35/60),80.00％(48/60),respectively.The positive rates of anti-3E antibody,anti-GP210 antibody,anti-SP1O0 antibody,anti PML antibody and anti-AMA-M2 antibody detected by CLIA were significantly higher than by ELISA(x2 =18.81,11.51,13.23,15.08,13.75,all P＜0.05).Concelusion Compared with ELISA,CLIA in the detection of related antibodies of patients with HCV can efficiently higher the positive rate of anti-HCV and autoantibody.
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Objective To systematically evaluate the value of chemiluminescence assay for detecting human immunodeficien-cy virus(HIV) antibodies as a HIV preliminary screening method .Methods The Chinese and English studies on chemilumines-cence assay for detecting HIV antibodies published by the databases of WanFang ,VIP ,CNKI ,CBM ,Pubmed and Web of Science were collected by computer retrieval and manual retrieval .The retrieval time limit was from the database establish to November 2016 .Two reviewers independently screened the literature ,extracted the data and assessed the methodological quality of included studies .Then the meta-analysis was performed by using Meta-disc1 .4 and Stata12 .0 software .Results A total of 8 studies invol-ving 26168 patients were included .The meta-analysis results showed that the pooled sensitivity of chemiluminescence assay for de-tecting HIV antibodies was 100% (95% CI:1 .00-1 .00) ,the pooled specificity was 100% (95% CI:0 .99-1 .00) ,the pooled posi-tive likelihood ratio was 237 .79(95% CI:80 .50-702 .42) ,the pooled negative likelihood ratio was 0 .00(95% CI:0 .00-0 .02) ,the diagnostic odds ratio(OR) was 48911 .05(95% CI:8257 .50-289711 .20) ,and the area under the curve(AUC) was 0 .9994(SE=0 .0002) .Conclusion Chemiluminescence assay can serve as a HIV preliminary screening method for promotion and application in clinic .
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Objective Detect the infection rate of herpes simplex virus (HSV ) by jointly using chemiluminescence assay and PCR ,and provides reference for clinical diagnosis .Methods The serum samples were collected from the pregnant women who had routine examination records in the hospital .Chemiluminescence assay was used to detect HSV IgM and IgG in those samples .Cervi‐cal secretions were collected from pregnant women with positive results and qualitatively tested for HSV DNA .Results The posi‐tive rate of HSV1 DNA was 0 .5% (7/1 422) ,the positive rate of HSV2 DNA was 1 .1% (16/1 422) .For pregnant women whose HSV IgM and IgG were both positive ,positive rate of HSV1 DNA was 0 .4% (4/1 008) and that of HSV2 DNA was 0 .6%(6/1 008);for those who only had HSV IgM positive ,the positive rate of HSV1 DNA was 0 .8% (1/130) ,and that of HSV2 DNA was 3 .1% (4/130);for those who only had HSV IgG positive ,the positive rate of HSV1 DNA was 0 .7% (2/284) ,that of HSV2 DNA was 2 .1% (6/284) .Among those three HSV antibody positive cases ,the difference in HSV1 DNA positive rate was not sta‐tistically significant(P>0 .05) ,while the difference in HSV2 DNA positive rate was statistically significant(P<0 .05) .Conclusion The test of HSV antibodies during pregnancy can be used as a routine test ,and HSV DNA test can be used as further test for those with HSV antibody positive ,which could improve the accuracy of diagnosis .Early screening ,detection ,and treatment are im‐portant for pregnant women with HSV infection .
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Objective To verify enzyme activity inhibition of a novel histone deacetylase inhibitor ( HDACi ) JZ005 using an HDACi chemiluminescence detection kit and a cell-based screening model .Methods The plasmid with p21 gene promoter elements and luciferase reporter gene was transfected into human embryonic kidney cells 293 , and the stable transfectants were established by G418 screening.Enzyme activity inhibition of JZ005 on histone deacetylases (HDACs) was verified by the HDACi chemiluminescence detection kit and the cell-based screening model .A well-known HDACi , tri-chostatin A ( TSA) was used as the positive control .MTT assay was used to detect the protection of rat H 9c2 myocardial cells suffering from CoCl 2-induced hypoxia and treated with different concentrations of JZ 005 .The expression of acetylated histone H3 protein of normal and CoCl 2-induced hypoxia H9c2 cells before and after JZ005 treatment was assayed by West-ern blotting while the effect of drug administration on apoptosis was detected by flow cytometry ( FCM) .Results An HDA-Ci cell-based screening system targeting the p21 gene promoter was ranging established .The JZ005, a HDACi, markedly suppressed the activity of HDACs by more than 50%with the concentration ranging from 50 to 400 μmol/L.JZ005 signifi-cantly protected H9c2 cells from hypoxia injury .Cell viability was increased by 38.33%,56.00% and 35.20% compared with control,accompanied by an enhanced acetylation level of histone H 3.JZ005(25,50 and 100 μmol/L) treatment sig-nificantly decreased the number of apoptotic cells (6.63%,10.56% and 8.89%) compared to control group (12.89%). Conclusion An HDACi cell-based screening system is successfully established .JZ005 effectively protects myocardial cells against hypoxia injury while enhancing the acetylation level of histone H 3.Our results indicate that JZ005 might be developed as a potential drug for hypoxia treatment .
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Objective To develop a chemiluminescence ( CL ) imaging DNA microarray method for simultaneous detection of seven rickettsiae.Methods Primers and probes were designed based on the specific sequence of seven rickettsia genomes.The probes were immobilized on the aldehyde modified glass surface to prepare DNA microarray for rickettsiae.The nucleic acids of the selected rickettsiae were amplified and labelled by multiplex PCR method, and then hybridized with microarray that was scanned after washing and chemiluminescence coloration, before the results were analyzed.Facilitated by the optimization of the multiplex PCR system, hybridization, and chemiluminescence imagination, we evaluated the specificity,sensitivity and reproducibility of the chip.The serial diluted nucleic acid of Rickettsia mooseri was detected using microarray and real-time PCR approach to compare the sensitivity of these two methods.Double-blind simulated samples were prepared to further evaluate the accuracy of the detection methods.Results One universal primer, four specific primers, one universal probe, and nine specific probes were selected.This DNA microarray demonstrated high specificity and sensitivity.The detection sensitivity of plasmid DNA and double-blind simulated sample DNA was 1.5 ×102 -3 ×103 copies per reaction and 103 -104 copies/μl.The detection results of real-time PCR method was consistent with the microarray, and the microarray possessed 10 fold lower detection sensitivities than the real-time PCR method.The coincidence rate of double-blind simulated sample detection was 100%.Conclusion A chemiluminescence imaging DNA microarray method for simultaneous detection of seven rickettsiae is established successfully,which can serve as a new high throuthput detection method for clinical diagnosis and epidemiological investigation of rickettsiae.
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Objective To investigate the relationship between extent of hepatic fibrosis and serum thyroid hormone levels in pa-tients with liver cirrhosis .Methods Chemiluminescence immunoassay technology was adopted to detect serum hyaluronic acid (HA),laminin(LN),collagen type Ⅳ (CIV),procollagen type Ⅲ (PC Ⅲ ),thyroid stimulating hormone(TSH),triiodothyronine (T3) ,thyroxine(T4) ,free thyroxin 3(FT3) and FT4 of 240 patients with liver fibrosis (liver fibrosis group) and 80 healthy people (control group) .Results In the control group ,serum HA ,LN ,CIV ,PCⅢ levels of healthy people in ≥45-year group were signifi-cantly higher than those in <45-year group ,and those in male group were obviously higher than female(P<0 .05) .Serum TSH , T3 ,T4 ,FT3 ,FT4 levels of healthy people in male group were significantly lower than female (P<0 .05) .With the increase of grade in Child-Pugh classification ,serum levels of hepatic fibrosis indexes of patients with liver fibrosis increased markedly (P<0 .05) , while their thyroid hormone levels significantly decreased (P<0 .05) ,especially in T3 and FT3 .Serum HA ,LN ,CIV ,PCⅢ levels of patients with A ,B or C grade were markedly higher than those in control group(P<0 .05) ,and serum T3 ,T4 ,FT3 ,FT4 levels of patients with B or C grade were obviously lower than those in control group (P<0 .05) .Conclusion The extent of liver fibrosis is correlated to serum thyroid hormone levels .
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Objective To analyze the correlation of real-time fluorescent quantitation PCR(FQ-PCR)for detecting HCV-RNA loading and the chemiluminescence immunoassay(CLIA)for detecting anti-HCV antibody.Methods 587 samples of anti-HCV an-tibody positive detected by CLIA were furteher detected HCV-RNA by FQ-PCR.Results Among 587 samples of anti-HCV anti-body positive by the CLIA screening,225 samples were HCV-RNA negative and 362 samples were HCV-RNA positive detected by FQ-PCR,and the positive rate was 61 .67%,moreover,which was positively correlated with the S/CO ratio detected by CLIA.Con-clusion The positive rate of HCV-RNA is positively correlated with the S/CO ratio detected by CLIA.The result of HCV-RNA can be predicted according to the S/CO ratio.
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Objective To evaluate the performance of the hypersensitive C-reactive protein(hs-CRP) rapid quantitative chemilu-minescent detection kit .Methods According to National Committee for Clinical Laboratory Standards (NCCLS) EP10-A2 docu-ment ,hs-CRP rapid quantitative chemiluminescent detection kit was employed to measure the CRP at low ,medium and high con-centration levels of quality control serum .Bias ,total imprecision and their slope rates ,intercepts ,carryover ,non-linearity and drift were calculated ,and its clinical acceptability was evaluated .Results Bias and total imprecision of hs-CRP rapid quantitative chemi-luminescent detection kit were within the allowable ranges ,the average values of slope rates ,intercepts ,carryover ,non-linearity and drift were 1 .005 7 ,0 .537 8 ,0 .789 6% ,0 .019 2 ,0 .036 0 ,respectively ,the differences showed no statistically significance ( P>0 .05) .Conclusion hs-CRP rapid quantitative chemiluminescent detection kit has good accuracy and precision ,stable perform-ance ,and consistent with the clinical testing requirements .
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Objective To explore the signification and method of Cut-off verification and gray zone setting in chemiluminescent assay.Methods NCCLS EP-12 A2 document defines that C50 is the analyte concentration of cut off value for immunology qualitative test and C5-C95 interval is the range of analyte concentration that yields 5％ positive results to 95％ positive results for immunology qualitativc test.The C50 and C5-C95 interval of HBeAg in ARCHITECT i2000 were worked out according to the cut off value provided by HBeAg reagent calibrated in ARCHITECT i2000,which were verified to approve the character declaimed by manufactory or not.Gray zone was set and the procedure of cut off verification and gray zone set in chemiluminescent were built; A set of quality control was detected 20 times with two different lot HBeAg reagent kits,S/CO was caculated and compared with t test.Results C50 and C5-C95 interval of reagent (lot 06087L100,96378HN00) were 0.171 PEI U/ml,0.125 PEI U/ml; ＞0.154 PEI U/ml to 0.188 PEI U/ml,0.119 PEI U/ml to ＜0.150 PEI U/ml,respectively.S/CO of negative quality control and positive quality control were (0.550 ±0.038),(2.422 ±0.084) and(0.334 ±0.063),(3.587 ±0.321),respectively.They all approved the character (the sensitivity at cut off was less than 0.5 PEI U/ml)declaimed by manufactory,and the results of S/CO between two lot kits were obvious difference (t =9.944,15.499,P ＜0.01).Conclusion C50 and C5-95 interval can be used to verify cut off value and set gray zone in chemiluminescent assay;They may vary in different lot reagents and they must be verified to approve the character declaimed by manufactory.