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Objective:Chemiluminescence immunoassay was used to detect the levels of anticardiolipin antibody (aCL) -IgA/IgG/IgM and anti-β2-glycoprotein Ⅰ antibody (aβ2GPⅠ) -IgA/IgG/IgM in healthy non-pregnant and pregnant women to explore the changes of antiphospholipid antibody in different pregnancy periods.Methods:This prospective study was conducted in Shandong Provincial Maternal and Child Health Care Hospital Affiliated to Qingdao University, involving normal pregnant women who underwent prenatal examination and healthy non-pregnant women with no history of adverse pregnancy who underwent progestational eugenic health examination from April 2020 to August 2021. The levels of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM were detected using BIO-FLASH chemiluminescence immunoassay analyzer and P95 as well as P99 were calculated, respectively. The difference in the six data between non-pregnant and pregnant women was compared using Mann-Whitney U test. Kruskal-Wallis H test was used to compare the change of each antibody in different pregnancy periods and Spearman correlation was used to analyze the correlation between different trimester and the levels of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM. Results:A total of 454 cases met the inclusion criteria, and 435 cases were included in the analysis after excluding 19 cases, among them 110 were non-pregnant women and 325 were pregnant women, including 110 cases in the first trimester (≤13 +6 weeks), 110 cases in the second trimester(14 +0-27 +6 weeks), and 105 cases in the third trimester (≥28 weeks). P99 value of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM in the non-pregnant women were 7.31, 14.70, 7.92, 3.58, 13.60, and 4.95 CU, which in the pregnant women were 5.90, 12.78, 5.70, 1.60, 10.65, and 3.90 CU, and were all lower than the cut-off value of 20 CU that given by the analyzer manufacturer. The levels of aCL-IgA/IgG/IgM, and aβ2GPⅠ-IgG/IgM in the pregnant women were significantly decreased comparing with the non-pregnant women [aCL-IgA: 1.90 CU (1.40-2.70 CU) vs 2.90 CU (2.20-3.83 CU), Z=－7.14; aCL-IgG: 3.00 CU (2.20-4.50 CU) vs 6.10 CU (4.20-7.83 CU), Z=－10.26; aCL-IgM: 1.40 CU (1.10-2.30 CU) vs 2.65 CU (2.08-3.73 CU), Z=－8.87; aβ2GPⅠ-IgG: 3.50 CU (2.60-4.90 CU) vs 4.75 CU (3.60-5.93 CU), Z=－5.45; aβ2GPⅠ-IgM: 0.70 CU (0.50-1.20 CU) vs 1.00 CU (0.60-1.53 CU) , Z=－3.73; all P<0.001]. The aCL-IgA level in the third trimester was higher than those in the first and second trimester (both P<0.05). The levels of aCL-IgG/IgM in the second trimester and aβ2GPⅠ- IgG in the second and third trimesters were significantly decreased than those in the first trimester (all P<0.05). Spearman analysis showed that aCL-IgG/IgM, aβ2GPⅠ-IgA/IgM had no significant correlation with the pregnancy period (the first, second and the third trimester) (all P>0.05). However, a weak correlation between the aCL-IgA, aβ2GPⅠ- IgG and the pregnancy period was observed ( r=0.28 and－0.49, both P<0.001) Conclusions:P99 value of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM levels in normal pregnant women and non-pregnant women are lower than the cut-off value of 20 CU given by the analyzer manufacturer. The levels of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgG/IgM during pregnancy are lower than those before pregnancy and fluctuate with the pregnancy period, but have no significant correlation with the pregnancy period. The clinical diagnosis of antiphospholipid syndrome should be made according to the cut-off values of aCL-IgA/IgG/IgM and aβ2GPⅠ-IgA/IgG/IgM determined by each laboratory.
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Objective Investigate the effect of neonatal hyperbilirubinemia on the detection of HBsAg by chemiluminescence and its elimination methods.Methods Case control study.The HBsAg in human serum was detected in 200 cases of hyperbilirubinemia neonates who were hospitalized in Beijing Children's Hospital,Capital Medical University from July 2015 to May 2016 and whose serum total bilirubin level exceeded 200 μmol/L.The positive serum was further detected by 16 200×g high-spoed centrifugation or blue light irradiation for 8 hours,and the results of re-assay of HBsAg were recorded.The retest positive serum wastested for HBV DNA load and checked the results of their mother's examination in HBV.136 adult serum samples with total bilirubin levels exceeding 200 μmol/L in the Peking University First Hospital,were taken as reference to compare the influence of hyperbilirubinemia between adults and newborns on the determination of HBsAg.Results The median level of serum total bilirubin in neonates was 259.0 μ mol/L (226.5,312.5);median level of indirect bilirubin 244.1 μmol / L(212.5,295.8).Median level of serum total bilirubin in adults 356.4 μmol/L(295.9,435.1);median level ofindirect bilirubin 137.1 μmol/L (107.8,172.7).The HBsAg test was negative in adults,11 cases (5.5％) were positive in newborns,their" HBV DNA load was less than＜100 IU/ml.Among them,9 have inoculated hepatitis B vaccine and 2 were unknown.10 of 11 mothers of infants were healthy and 1 was positive for HBsAg,HBeAb,HBcAb.2 of the 11 positive specimens turned negative of HBsAg after high-speed centrifugation.In addition to high speed centrifugation,4 cases turned negative after blue light irradiation.5 cases remained positive after high speed centrifugation and blue light irradiation.Conclusions Neonatal hyperbilirubinemia,which is different from that of adults,is mainly caused by indirectly bilirubin increased,which is one of the main reasons for false positive detection of HBsAg by chemiluminescence in neonates.High-speed centrifugation and blue light irradiation can eliminate the influence of serum indirect bilirubin on the detection of HBsAg to the greatest extent.
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Objective To investigate the effect of pseudodeficiency alleles on the newborn screening of glycogen storage disease typeⅡ(GSDⅡ) by using afluorometric enzymatic assay to determine acidα-glucosidase (GAA) activity in dried blood spot (DBS). Methods A total of 30507 newborns' DBSs, obtained from Newborn Screening Center of Xinhua Hospital Shanghai Jiao Tong University School of Medicine from May to December 2017, were screened for GSDⅡby fluorometric enzymatic assay of GAA activity. The suspected positive DBSs after the first and second screening were directly analyzed by Sanger sequencing of GAA to confirm the diagnosis. Retrospective analysis of 3172 controls without GSDⅡand 36 GSDⅡpatients were conducted to investigate the carrier status of pseudodeficiency alleles. Statistical analysis of frequency of pseudodeficiency alleles were carried out by Chi-square test or Fisher exact probability test. Results GAA activity of 30507 newborns showed a positively skewed distribution. Twenty-nine cases of newborns, suspected to be GSDⅡwere confirmed to be normal with genetic analysis of the original DBSs. Among the 29 suspected positive cases, 24 cases were homozygous for pseudodeficiency alleles c. [1726A/A; 2065A/A], and the other 5 cases were c. [1726G/A; 2065G/A] heterozygote. The frequency of c. 1726G>Ahomozygote in 3172 non-GSDⅡcontrols was 2.08％(66/3172), and c. 1726G>A homozygote occurred in allelic conjunction with c. 2065G>Ahomozygote. Frequency of c. [1726A; 2065A] haplotype in 3172 controls was 3.2％(206/6344). Frequency of c. [1726A/A;2065A/A] homozygote in 36 GSDⅡpatients (16.67％, 6/36) was significantly higher than that in non-GSDⅡcontrols(2.08％, 66/3172) (χ2=34.517, P<0.001). Conclusions Pseudodeficiency alleles show a high frequency in Chinese, which leads to a high false positive rate in the newborns screening of GSDⅡ.The afterword genetic analysis of the original DBS after the GAA activity screening could reduce the effect of pseudodeficiency alleles on the newborns screening of GSDⅡ.
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Objective@#To investigate the effect of pseudodeficiency alleles on the newborn screening of glycogen storage disease type Ⅱ(GSDⅡ) by using afluorometric enzymatic assay to determine acid α-glucosidase (GAA) activity in dried blood spot (DBS).@*Methods@#A total of 30 507 newborns′ DBSs, obtained from Newborn Screening Center of Xinhua Hospital Shanghai Jiao Tong University School of Medicine from May to December 2017, were screened for GSD Ⅱ by fluorometric enzymatic assay of GAA activity. The suspected positive DBSs after the first and second screening were directly analyzed by Sanger sequencing of GAA to confirm the diagnosis. Retrospective analysis of 3 172 controls without GSDⅡand 36 GSD Ⅱ patients were conducted to investigate the carrier status of pseudodeficiency alleles. Statistical analysis of frequency of pseudodeficiency alleles were carried out by Chi-square test or Fisher exact probability test.@*Results@#GAA activity of 30 507 newborns showed a positively skewed distribution.Twenty-nine cases of newborns, suspected to be GSDⅡwere confirmed to be normal with genetic analysis of the original DBSs. Among the 29 suspected positive cases, 24 cases were homozygous for pseudodeficiency alleles c.[1726A/A; 2065A/A], and the other 5 cases were c.[1726G/A; 2065G/A] heterozygote. The frequency of c.1726G>Ahomozygote in 3 172 non-GSD Ⅱcontrols was 2.08% (66/3 172), and c.1726G>A homozygote occurred in allelic conjunction with c.2065G>Ahomozygote. Frequency of c.[1726A; 2065A] haplotype in 3 172 controls was 3.2%(206/6 344). Frequency of c.[1726A/A; 2065A/A] homozygote in 36 GSDⅡpatients (16.67%, 6/36) was significantly higher than that in non-GSD Ⅱcontrols(2.08%, 66/3 172) (χ2=34.517, P<0.001).@*Conclusions@#Pseudodeficiency alleles show a high frequency in Chinese, which leads to a high false positive rate in the newborns screening of GSDⅡ.The afterword genetic analysis of the original DBS after the GAA activity screening could reduce the effect of pseudodeficiency alleles on the newborns screening of GSDⅡ.
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Objective@#To establish and evaluate a domestic Chemiluminescence immunoassay(CLIA) for detecting serum human epididymis protein4(HE4).@*Methods@#To establish a double antibody sandwich CLIA for the determination of serum HE4. Fluorescein isothiocyanate (FITC) and alkaline phosphatase (ALP) were used to label two different monoclonal antibodies of HE4. The separation could be realized by the magnetic particles coated with anti-FITC antibodies. Measure the relative light unit (RLU) after adding substrate solution, the RLU is proportional to the concentration of HE4. After evaluating the analytical performance including sensitivity, precision, accuracy, linearity, specificity under the optimized condition, this study compared the method with the commerical HE4 kits as well.@*Results@#The precision of with-in lot , and with-out lot are less than 4.0% and 5.0% respectively; Recovery is within 90.0%-110.0%; LoB is 5.0 pmol/L; functional sensitivity is 15.0 pmol/L; measure range is 15.0-1 500.0 pmol/L; the Correlation coefficient is bigger than 0.99 comparing with commercial kits. The AUC(area under curve) of three methods is 0.895, 0.900 and 0.896 respectively.@*Conclusions@#This study established a sensitive CLIA method with high repeatability and wide measure range; it has good correlation with commercial HE4 test kits.(Chin J Lab Med, 2018, 41: 615-620)
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Objective To explore the distribution characteristics of plasma renin concentration in patients with hypertension and the possibly methodological problems.Methods The subjects including 361 patients with hypertension[male: 184 cases, average age: (45.16±13.74)years old;female: 177 cases, average age: (51.04±12.68) years old]and 187 apparent healthy individuals[male: 92 cases, average age: (46.74±13.17)years old;female: 95 cases, average age: (47.33±13.18) years old]were recruited from Departments of Healthy Check-up and outpatients for hypertension in Tangshan Gongren Hospital. The plasma renin concentration was detected by chemiluminescence-Immunoassay.Results The plasma renin concentration shows log-transformed normal distribution both in healthy group and hypertension group. The range of plasma renin concentration in hypertension group is from 0.05 to 574.07 pg/ml, while that in apparent healthy group is from 3.24 to 120.40 pg/ml. The plasma renin concentration in both groups is higher in male than female (Hypertension t=2.19,P=0.029;Healthy people t=2.85,P=0.005). The average concentration of plasma renin in hypertension group is slightly higher, and the width of density distribution is larger in comparison with healthy group although there is no significant difference between them. However, the percentage of plasma renin abnormality was 26.59% (96/361) in hypertension group with 13.85%(50/361)of low renin subtype and 12.74%(46/361)of high renin subtype ConclusionsThe plasma renin concentration measured by Chemiluminescence-Immunoassay can be used as an effective tool for hypertension screening.
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Objective To study the changes of six sex hormones corresponding to the follicle growth during the normal menstrual cycle of Chinese women.Methods Thirty Chinese women with regular menstrual period and average age of (28.8±3.2) years were selected for the study by Peking Union Medical College Hospital in September,2010.Growth of follicles was monitored by using transvaginal sonography.Six sex hormones,including follicle-stimulating hormone (FSH),luteinizing hormone (LH),estradiol (E2),progesterone (P),testosterone (T),and prolactin (PRL) were measured by chemoluminescence immunoassay every day during a menstrual cycle.Nonparametric statistical analysis was used.ResultsMenstrual cycle of all the patients was in the range of 25 to 39 d,with mean of (29.5 ± 3.1) d.Length of follicular phase and luteal phase was 15.3 and 14.4 d,respectively.Number of days from antral follicle to emergence of dominant follicle,and from the latter to ovulation,was 6.2 and 8.9 d,respectively.Average diameter of preovulatory follicle was 19.33 mm.Both FSH and LH reached peak on the day before ovulation.P started to increase before ovulation and remained at a high plateau from 6th to 9th day after ovulation.Both PRL and T reached peak after ovulation,near the end of a menstrual cycle.Conclusions A small rise of LH and P emerges just 1 to 2 d before ovulation,indicating the maturing of follicle.PRL and T shows cyclic changes as follicle grows.Therefore,PRL and T levels should be measured in the early follicle phases in the clinical practice so that leading the impact of menstrual cycle minimal.
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Objective To compose and evaluate the measurement uncertainty of two kinds of chemiluminescence detection system using different methods.Methods The measurement uncertainty was composed by 4 different methods:(1) U 1％ was composed of within-run CV(CVw ％),between-run CV(CVB ％)and bias (CVBias ％);(2) U2％ was composed of CVB ％ and uncertainty of calibration (CVcal ％);(3) U3％ was composed of CVW％,CVs％ and CVcal％;(4) U4％ was composed of CVW％,CVB％,CVBias％ and CVcal％.The measurement uncertainty of Architect i2000SR system (Abbott,USA) and DXI800 system (Beckman,USA) was assessed.Pearson correlation analysis,Spearman correlation analysis,Paried t test and Mann-Whitney u test were performed to analyze the data.Results For Architect i2000SR system,U1％,U2％,U3％ and U4％ were significantly correlated (r=0.727-0.988,all P＜0.05),U3％ and U2％ were significantly different (t =6.88,P＜0.05),U4％ and U1％ were significantly different (t =6.21,P＜0.05).For DXI800 system,U1％,U2％,U3％ and U4％ were also significantly correlated (r =0.608-0.975,all P＜0.05),no significant difference was found between U3％ and U2％ (z=-1.33,P＞0.05),or between U4％ and U 1％ (z =-1.04,P＞ 0.05);the expanded measurement uncertainty was correlated with CVW％,CVB％,CVBias％(rs=0.653-0.912,all P＜0.05),but not with CVcal％(rs=0.548,P＞0.05).Conclusions For Architect i2000SR system,the fourth method is more proper to compose the measurement uncertainty (U4％).For DXI800 system,the first method is more appropriate (U1％).According to the contribution of different components to the measurement uncertainty,the measurement quality could be improved by reducing the imprecision and bias.
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Objective To investigate relationships between signal/cutoff (S/CO) ratios of antiHCV recombinant immunoblot assay (RIBA) and their positivity with different chemiluminescence immunoassay(CLIA) reagents.Methods A case-control study was performed.From March 2014 to March 2015,anti-HCV antibody was detected in 2 616 serum of outpatients and inpatients coming from Department of Clinical Laboratory,Qilu Hospital of Shandong University by three kinds of homemade CLIA reagents and one imported CLIA reagents.The positive samples were further tested by RIBA.The correlation between the positivity and the S/CO ratios was analyzed.The difference between different reagents were compared by x2 method.Results The predicted positivities of Shandong Laibo were 97.8％ and 33.3％ with S/CO ratio ≥ 26.8 and 1 to 26.8,respectively;The predicted positivities of Beijing Yuande were 96.7％ and 20％ with S/CO ratio ≥ 16.6 and 1 to 16.6,respectively;The predicted positivities of Beijing Kemei were 97.0％ and 9.8％ with S/CO ratio ≥ 16.7 and 1 to16.7,respectively;The predicted positivities of Abbott were 96.9％ and 12.8％ with S/CO ratio≥5 and 1 to 5,respectively.Conclusions Anti-HCV CLIA S/CO ratio and RIBA confirmatory test results have some relevance.Domestic reagents also can refer to import reagents determine the relationship between the positivity and the S/CO ratio.Different domestic reagent of positivity has different S/CO ratio.Although each reagent S/CO ratio to the same positivity has large difference,suggesting each manufacturer should set their products corresponding values according to the situation,providing reference for the clinical use of unit in result determination of the clinical trials.
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Objective To detect the changes of the NJ001 specific antigen expression before and after surgery, and evaluate whether the NJ001 specific antigen could be used as a serum biomarker for the diagnosis of pancreatic cancer.Methods With the method of sandwich ELISA, the serum samples from 85 pancreatic cancer patients, 22 pancreatic benign tumor and 40 healthy controls were detected respectively. The results of the NJ001 specific antigen in the serum samples from 85 pancreatic cancer patients were compared with CA19-9 detected by ECLIA.Results The positive rate of NJ001 for the pancreatic cancer group was obviously higher than that for the benign pancreatic tumor and health control groups[50.6%(43/85) vs 18.2%(4/22), χ2 =7.451, P0.05].The positive rate in the group of pancreatic cancer before surgery was higher than that after surgery[50.6%(43/85) vs 23.5%(20/85),χ2=13.341, P<0.05].In addition, the results from 85 pancreatic cancer patients showed the specificity of NJ001 specific antigen was up to 87.1%.Although the positive rate of NJ001 specific antigen for pancreatic cancer was lower than that of CA19-9[50.6%(43/85) vs 75.3%(64/85), χ2 =11.121, P<0.05], it was higher when they combined [ 85.9%( 73/85 ) ] .Conclusions It shows high positive rate of NJ001 specific antigen in the patients of pancreatic cancer in this study, which suggests that NJ001 specific antigen might be a potential valuable biomarker for the diagnosis of pancreatic cancer.
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Objective To establish a chemiluminecentdetection method ( CLIA ) of HCV IgG antibody for the detection of HCV infection and therefore lay a foundation for the research and development of testing kit.Methods Based upon the indirect ELISA method, the microwell plate was coated with HCV-NS3 and HCV-Core antigen expressed through gene engineering, and the anti-human IgG antibody was labeled with horse radish peroxidase.In this way, the chemiluminesent detection method of HCV IgG antibody was established.Meanwhile, the serum specimen of randomly selected 198 patients infected with HCV from No.302 Hospital of PLA and 222 blood donors, and the results were compared.Results The HCV-IgG antibody, a positive consistent rate of 99.0%( 196/198 ) , a negative consistent rate of 98.2%(218/222), and a total consistent rate of 98.6%(414/420) were found through testing 420 serum specimen with self-made agent and contrast agent.One HCV positive serum was repetitively tested with the self-made agent for 10 times, and a coefficient of variation ( CV) of less than 10% was found.Conclusion The chemiluminescent detection method of HCV IgG antibody is initially established, and the method, with an outstanding specificity and sensitivity, is applicable for screening blood donors, clinically detecting HCV infection as well as epidemiological survey.
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Objective To evaluate the detectability of HIV antigen-antibody in the window period of acute infection by three HIV antigen-antibody assays.Methods Twenty-two samples of HIV seroconversion serum panels and thirty-seven HIV acute infected plasm samples from our laboratory collected from cohort study of men who have sex with men between 2009 and 2011,were assayed by ECLIA,CLIA and ELISA methods.All assays were evaluated for the ability to detect HIV in the window period,and the sensitivity of each assay for acute samples was analyzed.Chi square test was used for statistical analysis.Results The ability of detecting HIV in the window period of each assay was different.For HIV seroconversion serum panels,the results of ECLIA and CLIA assays were consistent,and the window period was shortened at least 1 to 5 days compared with ELISA assay.For HIV acute samples,all were HIV positive by ECLIA or CLIA assay,but for ELISA assay,94.6％ was positive.For samples before seroconversion,ECLIA and CLIA assay had the same sensitivity (93.5％),which is superior to ELISA assay (71.0％) (x2 =5.14,P ＜0.05).Conclusion The ability of detecting HIV in the window period was different for each assay.The results of ECLIA and CLIA assay are consistent,superior to ELISA assay.