Objective It describes procedures for designing an analyzing a method comparison experiment for serum albumin(ALB) assay and determining the relative bias between two methods using split patient samples.Method According to the procedure described by the NCCL approved guideline: method comparison and bias estimation using patient samples; 40 patient samples were analyzed in 5 operating days, analyze each patient sample in duplicate using both BCG and BCP method. The duplicates were assessed for each method within the same run. The coefficient of correlation was calculated as well as the bia between the methods.Results In the patient′s serum sample, the bias between BCG and BCP methods were 1.8% at 50g/L or 5.5% at 40 g/L or 11.6% at 30 g/L for albumin. In the commercial quality control serum products, the bias between the two methods varied up to 36%.Conclusion In determination of the patient′s ALB, the bias between BCG and BCP goes up when the concentration of ALB is decreased. But the bias between the two methods varies up to 36% when the quality control serum is determined.

ObjectiveTo evaluate the performance of antinuclear antibody (ANA) detection in clinical laboratories.Methods There were 2 external quality assessments (EQA) scheme for nuclear antibody detection.The panel consisting of 5 samples was distributed.Each participant laborotory of the EQA program was required to report the ANA qualitative results,patterns,titers and anti-double strain DNA (dsDNA) antibody,anti-extractable nuclear antigen(ENA) antibody,the percent agreements of which were calculated respectively.ResultsThe number of laboratories performing ANA test with IIF increased from 77.6％ ( 149/192 )in 2006 to 82.2％ (342/416) in 2011,while the number of laboratories performing ANA test with ELISA was in the range of 14.5％ ( 53/365 ) and 16.0％ ( 52/326 ).The positive percent agreements of IIF was over 98％.The positive percent agreement of ELISA were all over 90％.IIF showed more satisfying positive percent agreements than ELISA every year.Over 90％ of the laboratories reported correct results for samples with granular ANA pattern except 0613 and 0624.Over 95％ of the laboratories reported correct results for samples with homogeneous ANA pattern.Two samples with centromere pattern were correctly detected by 88.5％ ( 161/182 ),79.0％ ( 147/186 ) of the laboratories in 2007,while the sample with centromere pattern was correctly detected by 98.4％ (299/304), which indicated an improvement in the detection of centromere pattern.In ANA positive results,the lowest percentage of the laboratories reporting the median result was 36％ (94/261),while the highest percentage was only 85.5％(224/262).The satisfied results of anti-ENA antibody were over 90％.And those of anti-dsDNA antibody was over 85％.ConclusionsIIF is the most common method for ANA screening in clinical laboratories.ELISA is also used in some laboratories.The two methods reported satisfying results in ANA test.The detection of anticentromere antibodies is improved.But the results of ANA titer reported are unsatisfactory. ANA detection in routine practice needs to be improved by standardization.

Objective Anti-RNase virus-like particles containing HCV RNA 5'-UTR were used as positive samples in national external quality assessment ( EQA ) to evaluate the competency of clinical Laboratories for the quantification of HCV RNA and analyze the possible problems of domestic kits. Methods The quality control samples with target values in EQA panels were distributed nationally twice by National Center of Clinical Laboratory (NCCL) to participating laboratories for the quantification of HCV RNA in 2008 and 2009. Each panel consisted of 5 samples. All participants were required to carry out the detection and to return results in expected time. Positive samples were virus-like particles which had been calibrated against the WHO HCV International Standard (NIBSC96/798)and the results of positive samples from participants should be in the range of target value of logarithm ± 0. 5. The 2nd panel in 2008 contained the common HCV genotypes and the 2nd panel in 2009 contained serial diluted samples of genotype 1b. The results of positive samples detected with 3 different lots reagent (21001,21078 and 21097) from the 2nd EQA in 2009 were statistically analyzed using the analysis of variance, then Dunnett'S T3 and Tamhane'S T2 were used if heterogeneity of variance was found. Results There was 390 participating laboratories in 2008 and 428/426 in 2009. The percentages of laboratories within the range of target value of logarithm ± 0. 5 for varied genotypes were different. The percentages of laboratories for 1b were more than 91%, for 2a were 93.7% and 74. 2% ,for6 were 83.3% and 80. 3%. The CVfor the low-level sample was higher than that for the high-level sample in the same year. The numbers of laboratories reporting false-negative samples in 2008and the 2nd in 2009 were 5, 1 and 10 respectively. Statistical differences were found among the results of four quality control serum samples using 3 different reagents( F = 288.23, 324. 79, 291.98 and 261.16,P ＜0. 01 ). Conclusion The competency for detecting low concentration samples and samples with genotype 2a or 6 needs to be improved.

Objective To explore the effects of general anesthesia combined thoracic paraverte-bral block on postoperative pain and fast track single-port video-assisted thoracoscopic surgery (VATS).Methods Thirty patients,including male 20 and female 10,received single-port VATS were randomly and equally divided into two groups:group C received general anesthesia only,and group T received ultrasound-guided thoracic paravertebral nerve block combined with general anesthe-sia.Both groups did not use the patient-controlled analgesia,if insufficient analgesia happened (rest-ing VAS scores>4),than used dezocine intravenously as additional analgesia (a single-dose 5-20 mg, no more than 120 mg per day).The Ramsay scores at 1,4,8,12 h after the surgery and the mechani-cal withdrawal threshold on the day before the surgery,at 4,8,12,24 h after the surgery were recor-ded.The first time of post-operation pain feedback,the consumption of dezocine in the first 24 h after surgery,the incidence rates of side effects,the first time off-bed and the hospital stays were also re-corded.Results Compared with group C,the Ramsay scores at 8,12 h postoperatively in group T significantly decreased (P <0.05),and the mechanical withdrawal threshold at 4,8 h postoperatively significantly increased (P <0.05).The first time of post-operation pain feedback in group T was sig-nificantly longer than group C (P <0.05).The consumption of dezocine in the first 24 h after surgery significantly decreased in group T (P <0.05).The first time off-bed and the hospital stays in group T were shorter than group C (P <0.05).Also,the incidence rates of nausea,vomiting in the first 24 h postoperatively were lower in group T (P < 0.05 ).Conclusion General anesthesia combined with single-injected thoracic paravertebral nerve block can effectively relieve the postoperative pain in pa-tients undergoing single-port VATS,reduce the consumption of opioids in the first 24 h postopera-tively,cutting down the occurring rates of adverse reactions,which was beneficial to early ambulate and shortened the hospital stays.

Objective To investigate the potential role of human cytomegalovirus lower matrix phosphoprotein 65 (HCMV pp65) in murine systemic lupus erythematosus (SLE).Methods The prokaryotic vector pET-28b and eukaryotic vector pcDNA 3.0 were constructed to express the HCMV pp65 protein.All the C57BL/6 mice were inoculated with pp65 eukaryotic vector intramuscularly five times at 2-week intervals and then were bled via the retro-orbital vein.Subsequently,indirect ELISA was used to evaluate the concentration of anti-pp65 IgG,anti-dsDNA and ANA.At the same time,IL-1 b,IL-6,and TNF-α were determined by competitive ELISA.Results The early onset of autoantibodies and an overexpression of IL-6 were observed in immunized male C57BL/6 mice.Conclusion HCMV pp65 triggers the deregulation of humoral immunity in C57BL/6 mice,which indicates that the immune responses induced by HCMV pp65 may be involved in the development of SLE.

Objective To evaluate the performance of HCV RNA detection in the first EQA program in 2012 and analyze possible problems in clinical laboratories.Methods The panel consisting of 5 samples was distributed to 927 laboratories.Each panel contains one negative sample and 4 positive samples,which were virus-like particles calibrated by international standard.The pere ent agreements of all the laboratories for qualitative and quantitative results were calculated.Genomic means (GM) and standard deviations (s) of all laboratories and each reagent were calculated.The overall GM and the GM of each reagent were compared with expected results and correlation curves were calculated.Results The percent agreements of sample 1211,1212,1213,1214 for qualitative results wcrc 99.5％ (403/405),98.5％ (400/406),100.0％ (405/405),100.0％ (406/406),respectively.The percent agreement of the negative sample was 99％ (401/405).The percent agreements of sample 1211,1212 and 1213 for quantitive results were similar,which were 93.8％ (549/585),92.3 ％ (541/586) and 94.5％ (554/586).However,the agreement of sample 1214 was only 87.7％ (514/586)and the agreement of sample 1214 for reagent A was 67.2％ (92/137).The overall GM agreed with expected results,while GMs of reagent C,E and G deviated from expected results.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent C were 4.22,3.56,5.16 and 5.90,respectively.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent E were 4.52,3.78,5.55 and 6.29,respectively.GMs of sample 1211,1212,1213 and 1214 reported by labs using reagent G were 4.83,4.36,5.72 and 6.56,respectively.Conclusions The overall results of HCV RNA qualitative and quantitative detection are satisfactory.However,some problems still exist,such as deviation of GM of some reagents,the interassay variability,systematic deviation and accidental deviation,which show that the quality of reagents should be improved.

Objective To evaluate the agreement of antinuclear antibody (ANA) titer reported in clinical laboratories and analyze possible problems in clinical laboratories.Methods Experiment survey.The panel consisting of 5 samples was distributed to 533 laboratories.Each panel contains one negative sample and 4 positive samples,which were from individuals of autoimmune disease.ANA titer system was divided into traditional titer system with two-fold dilution and titer system with 3.2 times dilution.Clinical laboratories were required to report the ANA titers and initial screening dilution according to the standard used in routine work.Results were expressed as median titers and range for acceptable performance.As to titer system with two-fold dilution,acceptable performance on proficiency testing was defined by a result equal to median titer ± two two-fold dilutions.While as to titer system with 3.2 times dilution,acceptable performance on proficiency testing was defined by a result equal to median titer ± 3.2 times dilutions.The laboratories percentages with acceptable performance were calculated to evaluate their agreements.Results 412 laboratories reported ANA titer results,of which 11.9％ (49/412)reported results with two-fold dilution titer system,88.1％ (363/412)reported results with 3.2 times dilution titer system.The median titers of sample 1211,1212,1213 and 1215 reported with two-fold titer system were 1 ∶ 640,1∶ 320,＞ 1 ∶ 1280 and1∶ 160,respectively.The agreement within the median ANA titer reported with two-fold dilution titer system ranged from 24.5％ (12/49) to 57.1％ (28/49) and the lowest percentage of the results within the acceptable limits was 87.8％ (43/49).The median titers of sample 1211,1212,1213 and 1215 reported with 3.2 times dilution titer system were 1 ∶ 1000,1∶ 1000,＞ 1 ∶ 3200 and 1 ∶ 320,respectively.The agreement within the median ANA titer reported with 3.2 times dilution titer system ranged from 63.3％ (31/49)to 83.7％ (41/49).The lowest percentage of the results within the acceptable limits was 98.0％ (48/49).Conclusions The results of ANA titer reported are unsatisfactory.Standardization for reagent,microscopy,procedure and result interpretation is necessary to improve the agreement of ANA titer report in different laboratories.

BACKGROUND: The participation of laboratories in external quality assessment (EQA) programs is required for the quality assurance of nucleic acid amplification of Chlamydia trachomatis. This study aimed to construct a new quality control (QC) material applicated in EQA of C. trachomatis PCR. METHODS: A QC material-HTB-SiHa cells transfected with a recombinant plasmid containing the cryptic plasmid sequence-was constructed for C. trachomatis PCR detection, and four different panels, each consisting of 4 positive samples with serial dilution of the constructed QC material and 1 negative sample, were distributed by the National Center for Clinical Laboratories among four groups of 275, 268, 317, and 304 participants across China from 2011 through 2012. A total of eight commercial kits were used for C. trachomatis PCR detection in participants. RESULTS: Nine laboratories reported false-positive results (0.9%). As the series dilution increased, the correct reporting of the data sets decreased; the lowest correct rate was 96.3% in the weakest positive samples (104 copies/mL). Eight laboratories reported false-positive results, and 42 laboratories reported false-negative results in the EQA detection of C. trachomatis. No significant differences were observed in the detection of the constructed C. trachomatis positive samples (97.9%, 98.5%, 100%, 98.5%; P=0.36) and negative samples (100%, 99.0%, 100%, 99.0%; P=0.764) using four commercial kits commonly used in China. CONCLUSIONS: The results of the EQA study indicated that the constructed material provides a noninfectious, stable control material with sufficient volume for PCR detection of C. trachomatis.
Cell Line ; Chlamydia Infections/diagnosis ; Chlamydia trachomatis/*genetics ; DNA, Bacterial/*analysis ; False Negative Reactions ; Humans ; Laboratories/*standards ; Plasmids/genetics/*metabolism ; Polymerase Chain Reaction/*standards ; Quality Control ; Reagent Kits, Diagnostic

Objective To develop a candidate reference method for the measurement of serum glucose based on isotope dilution liquid chromatography tandem mass spectrometry(ID-LC/MS/MS)Methods An internal standard [~(13)C_6]glucose was added to serum samples and equilibrated with endogenous glucose.Serum proteins were removed by a precipitation with anhydrous ethanol.Serum glucose and the internal standard were then reacted with 1-phenyl-3-methyl-5-pyrazolone and the formed derivatives were analyzed by liquid chromatography tandem mass spectrometry with multiple reaction monitoring(MRM).The method was calibrated with bracketing calibrators and serum glucose concentrations were calculated by comparing the peak area ratios of samples with that of the calibrators.Results The within-run,between-run and total coefficients of variation averaged 0.36%,0.47%and 0.61%,respectively.The analytical recoveries ranged from 99.0% to 100.9%.Results of analyzing the certified reference material SRM 965a showed an average biases of-0.20%.Conclusions An ID-LC/MS/MS method for measuring serum glucose has been developed.The method is highly precise and accurate and may be used as a candidate reference method.

Objective To exlore the influence of internal quality control and external quality control assessment(EQA) resulting from applicability of control samples in measurement of whole blood viscosity (WBV) through the analysis and comparison of applicability of 1 non-Newtonian fluid internal quality control sample in 3 viscometers. Methods Viscometer B, C and D were used to measure WBV of 30 blood samples in parallel under the shear rate(SR) of 1 s-1,30 s~(-1) and 200 s~(-1), then the blood SR-WBV curves of 3 viscometers were drawn according to the results. At the same time, viscometers B, C and D were used respectively to determine the WBV of control A 10 times in one day, then the control A SR-WBV curves were mapped. Three viscometers were used to measure the manufactory control samples and control A 5 times in one day for 4 days. Four groups of daily values of manufactory control samples and control A of each instrument were used to carry out F test to calculate whether 4 daily values are difference. Finally, the control A was dispensed in 49 laboratories nationwide chosen for measurement. On the basis of viscometer used, 20 laboratories were classified as group B, 20 laboratories were classified as group C and 9 laboratories were classified as group D. Then the data under SR of 1 s~(-1) were analyzed to calculate the coefficient of variation (CV) in the group. Results There was significant difference among the WBV of blood samples measured by the viscometers B, C and D. The results under SR of 1 s~(-1) declined in turn, and they were highest under SR of 30 s~(-1) followed by the values of viscometer D and B and they were (8.14±0.75), highest under SR of 30 s-1 followed by the values of viscometer B and D, and they were (7.35±0.07), daily values of manufactory control and control A of each instruments in four groups were compared. Under SR of 1 s~(-1), there was no difference between daily values of manufactory control and control A in viscometer B (F = 2.63, 1.37, P > 0.05), and there was no difference of daily values of manufactory control among viscometer C and D (F = 0.33,3. 14, P > 0.05), but significant daily difference existed when control A was tested by viscometer C and D (F = 5.76, 8.00, P < 0.05). Under SR of 30 s~(-1), there was no difference of daily values of manufactory control among 3 viscometers(F =0.31, 0.18, 2.26, P >0.05), and there was no difference of daily values of control A among 3 viscometers' (F = 1.03, 1.83, 2.40, P > 0.05); Under SR of 200 s~(-1), there was no difference of daily values of manufactory control among 3 viscometers (F =2.59, 0.68, 2.96, P > 0.05), and there was no difference of daily values of control A among 3 viscometers (F=2.31, 3.01, 2.28, P>0.05). When control A was tested under SR of 1 s~(-1) in 49 laboratories nationwide, the WBV values in groups of viscometer B, C and D were (18.47±1.30), (11.17±2.38), viscometer D and C were 63.75% and 21.3%. Conclusions Control A could fully mimic the properties of whole blood steadily on viscometer B, but partially mimic viscometer C and D, so the control A is most appropriate for viscometer B. Because current non-Newtonian fluid internal quality control could mimic rheological properties of whole blood under specifically conditions, laboratories should evaluate the consistent degree between control and whole blood, only the candidates which can mimic the properties of whole blood approximately could be chosen as quality control of WBV. When third-party control is chosen to be samples of EQA, its applicability should be in consideration. Pretest should be performed adequately to define applicability of third-party control, so as to reduce the difference among laboratories due to applicability of control and reflect detection quality of laboratories exactly.