Objective To discuss the improvement for the verification of inter-day precision of PS activity assay and the verification protocol for limits of quantitative of FⅧ:C and FⅨ:C assay,aiming at the problems in the performance verification of anticoagulant protein S(PS),coagulation factor Ⅷ activity(F Ⅷ:C)and coagulation factor Ⅸ activity(F Ⅸ:C)assay.Methods SYSMEX CN-6000 and supporting reagents were used,and low-and high-value quality control(QCs)were used as the study samples.Following the American Clinical and Laboratory Standards Institute(CLSI)document EP15-A3,an inter-day precision verification study of the PS activity assay was performed with three reagent preparation methods designed(specification-required method,instrument's manual method and improved method).The specification-required method was carried out completely according to the specification,and the instrument manual method involved allowing the required reagents to stand in the device for 30 minutes based on the specification-required method,and the improved method mixed and dispensed the reagents needed to be based on the specification-required method.To study the bottle variation of the same batch of reagents for the same sample,the acceptable range of external quality assessment(EQA)of the National Center for Clinical Laboratories(NCCL)was used as the evaluation standard.Following the CLSI EP25 document,the PS activity assay reagent onboard stability verification was performed with the specification-required method and the improved method.Following the CLSI EP17-A2 document,WS/T 514-2017"Establishment and Verification of Detection Capability for Clinical Laboratory Measurement Procedures"and the International Committee for Standardization in Hematology(ICSH)guidelines,the LoQ for FVIII:C and FIX:C assays was verified.The verification results were passed if the requirements of the specification were met.Results The verification result of inter-day precision(CVWL:12.9%～21.6%)of PS activity assay according to the specification method and the instrumental manual method exceeded the requirements of the specification(<10%CVWL at high levels,<20%CVWL at low levels).The results of inter-day precision verification with the improved method(CVWL:2.9%and 4.5%)were consistent with the requirements of the specification.The bottle variation exceeded the acceptable range of EQA.The improved method corrected the reagent on-board stability verification results of reagents(relative deviations of-4.24%～9.97%)by the specifications(high levels<10%,low levels<20%).The LoQ verification results for FⅧ:C and FⅨ:C assay were by the product specifications(FⅧ:C:0.75%～1.46%and 0.74%～1.40%;FⅨ:C:0.71%～1.27%and 0.70%～1.32%).Conclusion An improved method to improve the inter-day precision of PS activity detection is provided,and LoQ verification protocol for F Ⅷ:C and FⅨ:C assay is provided for clinical laboratory reference.

Objective To discuss the improvement for the verification of inter-day precision of PS activity assay and the verification protocol for limits of quantitative of FⅧ:C and FⅨ:C assay,aiming at the problems in the performance verification of anticoagulant protein S(PS),coagulation factor Ⅷ activity(F Ⅷ:C)and coagulation factor Ⅸ activity(F Ⅸ:C)assay.Methods SYSMEX CN-6000 and supporting reagents were used,and low-and high-value quality control(QCs)were used as the study samples.Following the American Clinical and Laboratory Standards Institute(CLSI)document EP15-A3,an inter-day precision verification study of the PS activity assay was performed with three reagent preparation methods designed(specification-required method,instrument's manual method and improved method).The specification-required method was carried out completely according to the specification,and the instrument manual method involved allowing the required reagents to stand in the device for 30 minutes based on the specification-required method,and the improved method mixed and dispensed the reagents needed to be based on the specification-required method.To study the bottle variation of the same batch of reagents for the same sample,the acceptable range of external quality assessment(EQA)of the National Center for Clinical Laboratories(NCCL)was used as the evaluation standard.Following the CLSI EP25 document,the PS activity assay reagent onboard stability verification was performed with the specification-required method and the improved method.Following the CLSI EP17-A2 document,WS/T 514-2017"Establishment and Verification of Detection Capability for Clinical Laboratory Measurement Procedures"and the International Committee for Standardization in Hematology(ICSH)guidelines,the LoQ for FVIII:C and FIX:C assays was verified.The verification results were passed if the requirements of the specification were met.Results The verification result of inter-day precision(CVWL:12.9%～21.6%)of PS activity assay according to the specification method and the instrumental manual method exceeded the requirements of the specification(<10%CVWL at high levels,<20%CVWL at low levels).The results of inter-day precision verification with the improved method(CVWL:2.9%and 4.5%)were consistent with the requirements of the specification.The bottle variation exceeded the acceptable range of EQA.The improved method corrected the reagent on-board stability verification results of reagents(relative deviations of-4.24%～9.97%)by the specifications(high levels<10%,low levels<20%).The LoQ verification results for FⅧ:C and FⅨ:C assay were by the product specifications(FⅧ:C:0.75%～1.46%and 0.74%～1.40%;FⅨ:C:0.71%～1.27%and 0.70%～1.32%).Conclusion An improved method to improve the inter-day precision of PS activity detection is provided,and LoQ verification protocol for F Ⅷ:C and FⅨ:C assay is provided for clinical laboratory reference.

Objective:To investigate the analytical performance verification protocols and performance specifications of CD34+cell enumeration by flow cytometry for clinical laboratories.Methods:According to international guidelines and National Health Standard of China, we designed the performance verification protocols of CD34 +cell enumeration (including percent count and absolute count) by flow cytometry. Four quality assessment materials, three leukapheresis products and three samples of peripheral blood were selected to verify the precision, linearity, carryover, trueness and accuracy of FACSCanto Ⅱ measurement system, and the assessment criterion was set according to the detection technologies of clinical laboratories. Results:The CVs of intra-run precision of percent count and absolute count were 2.5% to 8.9% and 3.0% to 9.0%; the CVs of inter-run precision were 2.8% to 10.5% and 3.8% to 9.9%, respectively. The slopes of linearity regression equation of low range (3.6/μl to 123.6/μl) and high range (113.2/μl to 1196.3/μl) were 0.993 2 and 0.965 2, and R2 were 0.999 6 and 0.993 9, and the biases were -8.67% to 0.22%. The carryover of percent and absolute count were 0.07% and 0.00%. When percent count≤0.2% or absolute count≤20/μl, the absolute biases of trueness were in the range of ±0.006% or ±0.5/μl, and the absolute biases of accuracy were in the range of ±0.02% or ±0.9/μl; when percent count>0.2% or absolute count>20/μl, the relative biases of trueness were in the range of ±5.65%, and the relative biases of accuracy were in the range of ±8.19%. The verification results met the assessment criterion set in this study. Conclusions:The performance verification protocols and assessment criterion formulated in this study not only conform to the recommendations of domestic and foreign guidelines, but also conform to state of the detection technologies of native clinical laboratories, which can be taken as a reference of performance verification for clinical laboratories.

Objective: To assess the association of single nucleotide polymorphisms (SNPs) in SLCO1B3 gene with prognosis of breast cancer (BC) patients treated with neoadjuvant chemotherapy of TA regimen (taxane and antharcycline drugs). Methods: 439 female BC patients were recruited and treated with neoadjuvant chemotherapy of TA regimen. A blood sample (2 ml) of peripheral blood was collected from each patient before chemotherapy. Tagging SNPs (tag-SNPs) were selected. We investigated the association of tag-SNPs with prognosis, by Sequenom Mass ARRAY system platform, characterizing tag-SNPs. The hazard ratio (HR) and 95% confidence interval (CI) for progression or death were calculated by multivariable-adjusted Cox regression model. Results: Seven tag-SNPs (rs11045689, rs200104106, rs3764006, rs3834935, rs4149117, rs7305323 and rs73241801) were selected for study. Compared with individuals carrying the rs11045689 GG genotype, individuals carrying rs11045689 AA genotype performed worse PFS and OS, with the HR (95% CI) for progression being 1.39 (1.11~1.75) and the HR (95% CI) for death being 1.38 (1.04~1.83). Compared with individuals carrying the rs73241801 CC genotype, individuals carrying rs73241801 TT genotype performed better OS (P=0.041), with the HR (95% CI) for death being 0.65 (0.44~0.94). The number of risk allele was significantly associated with PFS (P=0.012) and OS (P=0.017) of BC patients by accumulation analysis. Compared with individuals carrying one or less than one risk allele, individuals carrying four risk alleles performed worse PFS and OS, with the HR (95% CI) for progression being 1.37 (1.09~1.72) and the HR (95% CI) for death being 1.36 (1.02~1.81). Conclusion The variations of rs11045689 and rs73241801 in SLCO1B3 gene were significantly associated with prognosis of BC patients treated with neoadjuvant chemotherapy of TA regimen, which might serve as biomarkers for predicting prognosis of BC patients treated with neoadjuvant chemotherapy.

Objective To assess the association of single nucleotide polymorphisms ( SNPs) in SLCO1B3 gene with prognosis of breast cancer (BC) patients treated with neoadjuvant chemotherapy of TA regimen (taxane and antharcycline drugs). Methods 439 female BC patients were recruited and treated with neoadjuvant chemotherapy of TA regimen.A blood sample (2 ml) of peripheral blood was collected from each patient before chemotherapy. Tagging SNPs (tag?SNPs) were selected. We investigated the association of tag?SNPs with prognosis, by Sequenom Mass ARRAY system platform, characterizing tag?SNPs. The hazard ratio ( HR ) and 95% confidence interval ( CI ) for progression or death were calculated by multivariable?adjusted Cox regression model. Results Seven tag?SNPs ( rs11045689, rs200104106, rs3764006, rs3834935, rs4149117, rs7305323 and rs73241801) were selected for study. Compared with individuals carrying the rs11045689 GG genotype, individuals carrying rs11045689 AA genotype performed worse PFS and OS, with the HR (95% CI) for progression being 1.39 (1.11～1.75) and the HR (95% CI) for death being 1.38 ( 1.04～1.83). Compared with individuals carrying the rs73241801 CC genotype, individuals carrying rs73241801 TT genotype performed better OS (P＝0.041), with the HR (95% CI) for death being 0.65 (0.44～0.94). The number of risk allele was significantly associated with PFS (P＝0.012) and OS (P＝0.017) of BC patients by accumulation analysis. Compared with individuals carrying one or less than one risk allele, individuals carrying four risk alleles performed worse PFS and OS, with the HR (95%CI) for progression being 1.37 (1.09～1.72) and the HR ( 95% CI) for death being 1.36 (1.02～1.81). Conclusion The variations of rs11045689 and rs73241801 in SLCO1B3 gene were significantly associated with prognosis of BC patients treated with neoadjuvant chemotherapy of TA regimen, which might serve as biomarkers for predicting prognosis of BC patients treated with neoadjuvant chemotherapy.

Objective To assess the association of single nucleotide polymorphisms ( SNPs) in SLCO1B3 gene with prognosis of breast cancer (BC) patients treated with neoadjuvant chemotherapy of TA regimen (taxane and antharcycline drugs). Methods 439 female BC patients were recruited and treated with neoadjuvant chemotherapy of TA regimen.A blood sample (2 ml) of peripheral blood was collected from each patient before chemotherapy. Tagging SNPs (tag?SNPs) were selected. We investigated the association of tag?SNPs with prognosis, by Sequenom Mass ARRAY system platform, characterizing tag?SNPs. The hazard ratio ( HR ) and 95% confidence interval ( CI ) for progression or death were calculated by multivariable?adjusted Cox regression model. Results Seven tag?SNPs ( rs11045689, rs200104106, rs3764006, rs3834935, rs4149117, rs7305323 and rs73241801) were selected for study. Compared with individuals carrying the rs11045689 GG genotype, individuals carrying rs11045689 AA genotype performed worse PFS and OS, with the HR (95% CI) for progression being 1.39 (1.11～1.75) and the HR (95% CI) for death being 1.38 ( 1.04～1.83). Compared with individuals carrying the rs73241801 CC genotype, individuals carrying rs73241801 TT genotype performed better OS (P＝0.041), with the HR (95% CI) for death being 0.65 (0.44～0.94). The number of risk allele was significantly associated with PFS (P＝0.012) and OS (P＝0.017) of BC patients by accumulation analysis. Compared with individuals carrying one or less than one risk allele, individuals carrying four risk alleles performed worse PFS and OS, with the HR (95%CI) for progression being 1.37 (1.09～1.72) and the HR ( 95% CI) for death being 1.36 (1.02～1.81). Conclusion The variations of rs11045689 and rs73241801 in SLCO1B3 gene were significantly associated with prognosis of BC patients treated with neoadjuvant chemotherapy of TA regimen, which might serve as biomarkers for predicting prognosis of BC patients treated with neoadjuvant chemotherapy.

The pediatric reference intervals in clinical laboratory play an important role in diagnosis of illness,therapeutic monitoring,prediction of prognosis and health evaluation.Compared with establishing reference interval for adults,there are more challenges to establish pediatric reference intervals.Therefore,the procedure and key technologies of direct method and indirect method are stated based on the characteristics of children population and pediatric,by which to define,transfer and validate pediatric reference intervals.This study will provide systematically methodological ideas for clinical laboratories to establish pediatric reference intervals.

Objective To prepare and evaluate the reference materials for plasma von Willebrand Factor antigen testing with fresh frozen plasma.Methods The candidates were prepared by low temperature centrifugation in 5 different concentration levels.The homogeneity and stability of the preparation was evaluated according to the ISO Guide35 and CNAS-GL03.The comparability between STAGO and IL system was evaluated according to the WS/T 356-2011.Then the preparations were characterized by six laboratories with the Secondary Coagulation Standard established by NIBSC(SSCLOT4).Results Homogeneity evaluation of the preparation showed that there was no statistically significant difference between the groups (P >0.05),the F values of factor analysis of variance were 0.317~0.844,the uncertainty range was 1.01% ~2.06%.A linear regression based on stability evaluation indicated that the linear trend (within 24 weeks)was insignificant (P >0.05). The uncertainty range of long-term (within 24 weeks)stability was 0.79% ~ 1.20%.The results of the preparations on STAGO and IL system were comparable.The certificated values of the candidates were range from 12.2% to 138.9% with uncertainties were 0.06%~0.09%,respectively.The range of combined standard uncertainty was 0.03% ~ 0.16% while the expanded uncertainty was 2.2%~6.7%.Conclusion The reference materials for von Willebrand Factor antigen testing were stable and homogenous with comparability between STAGO and IL.The method of characterization was accurate and reliable.

Objective To investigate current status and problems of coagulation factor Ⅷ and Ⅸ assay in domestic laboratories so as to provide the reference for implementing the standardization and quality improvement.Methods A questionnaire survey was carried out in 76 laboratories,and quality control materials were distributed to 54 laboratories for activity assay.The questionnaire information was analyzed statistically.Test results of quality control materials were classified into three groups according to the reagents and the ranked grading analysis were used to evaluate the performance.Results This research was investigative study.The amount of sample was less than 30 per month in 72％ (52/72)of laboratories.The frequencies of calibration were different,and 33％ (24/72) of laboratories did not perform calibration in a different assay batch.39％ (28/72)of laboratories did not run internal quality control,and about 21％ (15/ 72) of laboratories just performed the normal level quality control.Individual laboratories showed a high cumulative CV (＞ 30％) of intemal quality control.For normal FⅧ and FⅨ control materials,the CV of results were 11.3％-18.2％ and 11.3％-17.9％ respectively as well as 15.3％-20.3％ and 19.5％-21％ for abnormal.Of the three groups,the proportions of laboratories which the FⅧ test results out with consensus were18％,24％ and 22％ as well as 20％,24％ and 28％ for FⅨ.Conclusions The key requirements for quality control of coagulation factors active assay remain to be addressed and implemented.The repeatability and comparability in some laboratories are not satisfactory to meet the clinical needs.With the purpose of promoting quality improvement,we need to develop guidelines,organize related training and establish a national external quality assessment scheme.

Objective To verify the reference interval for D-dimer assay and analyze the influence of age and gender on the ref-erence interval.Methods Inclusion criteria for reference individuals were established.60 healthy males and 63 females were enrolled and divided to three groups by age,including 20 to 39 years old group (20 males and 20 females),40 to 59 years old group (20 males and 23 females)and above 60 years old group (20 males and 20 females).Blood samples were drawn in cit-rate sodium anticoagulated tubes and D-dimer concentration was determined by three different coagulation analyzers using o-riginal reagents.According to CLSI guideline C-28-A3,the reference interval for each measurement system from reagent manufacturer was verified and the difference of D-dimer concentration between different age-group and sex-group was ana-lyzed using non-parameters tests.Results All reference intervals were verified for people under age 40,while one reference interval cannot be verified for people from 40 to 59 years old as same as one for people above 60 years old.D-dimer concen-tration increased with age and there was significantly different between 20~39 years old group and 40~59 years old group or above 60 years old group(P<0.05).There was only a significant difference between sex-group for people under age 60(P<0.05).Conclusion D-dimer concentration was associated with age and sex.For people under age 40,the reference inter-val from reagent manufacture can be verified and directly used in laboratory,while for people above age 60,the reference in-terval from reagent manufacture cannot be verified.The cause should be investigated and a new reference interval should be established separately when necessary.