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Electrophoresis technique, established in the mid-20th century, has derived many technical branches over many years of development, which plays an important role in clinical laboratory and scientific researches. The automation, intelligence, and miniaturization will promote continuous progress of electrophoresis technology in assisting the diagnosis, treatment and mechanism study of clinical diseases.
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Objective:To compare the distribution differences of serum protein electrophoresis (SPE) among different gender and age individuals, and to explore the clinical application of SPE screening monoclonal gammopathy.Methods:A retrospective analysis was conducted based on the SPE results obtained from 533 989 cases enrolled from January 2018 to December 2019 at Zhongshan Hospital Affiliated to Fudan University. Among these patients, 435 479 inpatients were from departments of hematology, nephrology, spinal surgery, endocrinology, and rheumatology and immunology; and 98 510 were apparently healthy individuals. The distributions of albumin, α1 globulin, α2 globulin, β1 globulin, β2 globulin and γ globulin in different gender and age groups (≤20, 21-30, 31-40, 41-50, 51-60, 61-70, 71-80, 81-90, ≥91 years old) were compared. A total of 10 014 cases were selected by immunofixation electrophoresis (IFE). The positive detection rates of different SPE bands and IFE bands were analyzed. The sensitivity and specificity of SPE methods were determined according to IFE results as the gold standard.Results:No significant difference was examined in the proportion of SPE bands between different genders ( P>0.05). There were statistically significant differences in the proportion of albumin bands between apparently healthy individuals and hospitalized patients at different ages (apparently healthy individuals: F=5.12, P<0.05, inpatients: F=4.18, P<0.05), and all of them decreased with the increase of age. The proportion of γ globulin bands increased with age (apparently healthy individuals: F=1.34, P<0.05; inpatients: F=1.24, P<0.05). The sensitivity of SPE was 69% (2 098/3 051), and the specificity was 97% (6 721/6 963). Compared with IFE method, the positive detection rate of monoclonal gammopathy was significantly different (χ2=5 049.94, P<0.05). The positive rate of monoclonal gammopathy in γ globulin region (21.11%, 2 114/10 014) was higher than that in β globulin region (3.28%, 328/10 014) (χ2=90.74, P<0.05) and β-γ globulin region (1.63%, 163/10 014) (χ2=44.34, P<0.05). IgG and IgM bands are common in γ globulin region. Among them, IgG-κ type accounted for 94.1% (995/1 058), IgG-λ type accounted for 94.8% (690/728), IgM-κ type accounted for 89.2% (222/249), IgM-λ accounted for 83.8% (62/74). IgA bands are common in β region, of which IgA-κ accounted for 49.8% (103/207) and IgA-λ accounted for 51.6% (149/289). The positive rate of monoclonal gammopathy of IgG-κ type was the highest (10.57%, 1 058/10 014), and the positive rate of monoclonal gammopathy of IgM-λ type was the lowest (0.74%, 74/10 014). Conclusions:With increasing age, the proportion of albumin band in SPE decreased and the proportion of γ globulin band increased. IgG and IgM type monoclonal gammopathy is mostly found in the gamma region, with a higher detection rate in IgG type. IgA type monoclonal gammopathy is mostly found in the β region, with a lower detection rate.
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Objective:To investigate the characteristics and distribution of monoclonal gammopathy in clinical patients.Methods:A total of 936 171 patients (508 449 males and 427 722 females) who received capillary zone electrophoresis in Zhongshan Hospital Affiliated to Fudan University from January 2012 to December 2021 were selected, from which 14 945 patients with abnormal bands were screened as the study subjects, including 10 173 males and 4 772 females and the age 21-102 (65±13) years old. According to the age, patients were divided into 8 groups: 21-30 years old (168 cases), 31-40 years old (405 cases), 41-50 years old (1 326 cases), 51-60 years old (3 068 cases), 61-70 years old (4 985 cases), 71-80 years old (3 288 cases), 81-90 years old (1 519 cases), and≥91 years old (186 cases). The diagnostic results of the 14 945 patients with abnormal bands were collected and were divided into tumor group (5 196 cases) and non-tumor disease group (9 749 cases) according to the presence of tumor. The distribution of abnormal bands in different gender, age, and disease groups were retrospectively analyzed. Among the 14 945 patients, 4 988 cases underwent immunofixation electrophoresis, excluding 336 negative cases and 412 cases of double clonal bands reaction, and 4 240 patients with monoclonal immunoglobulin (M protein) reaction were selected as the study subjects, including 2 794 males and 1 446 females aged 21-102 (67±12) years old. They were divided into 8 groups according to the age: 21-30 years old (18 cases), 31-40 years old (91 cases), 41-50 years old (364 cases), 51-60 years old (862 cases), 61-70 years old (1 455 cases), 71-80 years old (904 cases), 81-90 years old (486 cases), and≥91 years old (60 cases). The diagnostic results and immunoglobin subtypes (IgA-κ, IgA-λ, IgG-κ, IgG-λ, IgM-κ, IgM-λ, κ, λ) of patients were collected, and the distribution of monoclonal gammopathy in different gender, age and disease groups were retrospectively analyzed.Results:Among 936 171 patients, 14 945 cases showed abnormal bands in electropherograms with a detection rate of 1.60%; the detection rates of abnormal bands in males and females were 2.00% (10 173/508 449) and 1.12% (4 772/427 722), respectively, with a statistically significant difference ( P<0.01). There was a significant difference in the detection rate of abnormal bands among different age groups ( P<0.01); among them, the highest detection rate of abnormal band in group of ≥91 years old was 5.98%, and the ratio of male to female was 1.67∶1. Among the 14 945 cases of abnormal bands, patients aged 51-60, 61-70 and 71-80 accounted for 20.53% (3 068 cases), 33.36% (4 985 cases) and 22.00% (3 288 cases), respectively, and the differences among the age groups were statistically significant (χ 2=115.82, P<0.01). In the tumor group, the top 3 tumors with abnormal bands were plasmacytoma with 1 123 cases, lymphoma with 289 cases, and leukemia with 49 cases. The detection rate of abnormal bands in electropherograms of plasmacytoma was 89.92% (1 123/1 249), which was higher than that in lymphoma and leukemia [6.73% (289/4 296) and 6.40% (49/766), respectively, P<0.01]. Among 4 240 patients with positive M protein, the proportion of 51-60, 61-70 and 71-80 years old patients were 20.33% (862/4 240), 34.32% (1 455/4 240) and 21.32% (904/4 240), respectively, and the differences among age groups were statistically significant ( P<0.01). The results of M protein types showed that the proportion of IgG-κ type was the highest in both genders, with 32.28% (902/2 794) in males and 34.30% (496/1 446) in females. In the 21-30, 31-40, and 41-50 age groups, the proportion of IgG-λ was the highest, which were 38.89% (7/18), 36.26% (33/91) and 34.07% (124/364) in these groups respectively. However, the proportions of IgG-κ were the highest in either of the 51-60, 61-70, 71-80, 81-90 and ≥91 years old groups, which were 33.87% (292/862), 34.16% (497/1 455), 31.53% (285/904), 34.57% (168/486), 28.33% (17/60), respectively, and the differences among all age groups and gender groups had statistical significance ( P<0.01). Among patients with positive M protein in the tumor group, plasmacytoma accounted for 14.22% (603/4 240), followed by lymphoma 6.30% (267/4 240); among non-tumor diseases, M proteinemia accounted for the highest proportion (7.24%, 307/4 240), followed by pulmonary infection (5.47%, 232/4 240). Conclusions:The detection rate of abnormal bands in capillary zone electrophoresis may increase with age, and is higher in males than in females in the same age group; different malignant tumor diseases can also show abnormal bands in capillary zone electrophoresis, but they are still mainly hematological tumors. Among the positive results of M protein, 61-70 years old group accounts for the highest proportion; the most common type of monoclonal gammopathy is IgG type; in the age group of 21-50 years, the proportion of IgG-λ type is the highest; in the group of >50 years old, the proportion of IgG-κ type is the highest; in the diagnosis of positive monoclonal gammopathy, the top 3 diseases are all hematological diseases, including plasmacytoma, monoclonal gammopathies and lymphoma.
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Clinical application of mass spectrometry technology has attracted the attention of clinical laboratory experts due to its high sensitivity, high specificity, and capacities of simultaneous detection of multiple compounds. In recent years, mass spectrometry technology has made significant achievements in the fields of identification of pathogenic microorganism, detection of trace elements and heavy metals, small molecule hormones, vitamins, amino acids, peptides and proteins, as well as therapeutic drug monitoring (TDM) and poisoning drugs screening. In order to further clarify the opportunities and challenges brought by this complex mass spectrometry technology in the field of clinical laboratory, the Chinese Journal of Laboratory Medicine invited experts and scholars of laboratory medicine to share their experience and opinions on related items focusing on the positioning of mass spectrometry technology in the clinical laboratory, the development and improvement of the clinical laboratory by mass spectrometry technology, the challenges of interpreting mass spectrometry test results, the challenges of operating and managing clinical mass spectrometry laboratories, and ways of improving the application of clinical mass spectrometry laboratories with this technology. Agreement was achieved in that the introduction of mass spectrometry technology into the clinical laboratory could bring new directions and opportunities for clinical testing and research, and also is associated with a series of challenges such as the difficulty of sample pretreatment, the high cost and complexity of mass spectrometry technology, the complexity of data processing and interpretation, the lack of standards and norms, and the issue of determining the price of mass spectrometry examinations.
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Objective:To investigate the correlation between gender, age, blood collection time, season and changes in cTnT concentration.Methods:In this study, 3548 patients (non-cardiovascular diseases) in Zhongshan Hospital of Fudan University were selected from 1 January to 31 December 2019. The basic data of the patients were collected, including gender, age, time of blood collection, medical history, clinical diagnosis, and results of cTnT testing. 1 840 males and 1 708 females were finally enrolled, with an age distribution of 65 (53, 75) years. The distribution of the data was assessed using the Kolmogorov-Smirnov (K-S) test, where non-normally distributed data were expressed as M( Q1, Q3). The Mann-Whitney U-test was used to compare cTnT concentrations between men and women, and to analyse the influence of gender on cTnT results. The Kruskal-Wallis test was used to compare cTnT levels between gender groups, to analyse the correlation between different times of blood collection, seasons, and other factors and cTnT concentrations. Result:cTnT concentrations increased with age in both males and females over the age of 60 years. cTnT levels were highest in individuals over the age of 90 years (0.028 ng/ml in males and 0.018 ng/ml in females). cTnT levels were higher in males (0.012 ng/ml) than in females (0.009 ng/ml) in all age groups ( H=6.340, P<0.01). The concentrations of cTnT varied at different time points of blood collection. In both males and females, cTnT concentrations reached a maximum at 8:00 and 13:00 (0.013 ng/ml and 0.012 ng/ml, respectively). Analysis of the physiological effect of season on cTnT secretion showed that cTnT levels were generally higher in spring and winter(0.012 ng/ml) than in summer and autumn(0.010 ng/ml). Conclusions:cTnT concentration is influenced by gender, age, time of blood collection and season. When analysing cTnT results in clinical practice, the gender and age of the individual should be taken into account, as well as the time point of blood collection and seasonal factors.
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The rapid development of laboratory medicine contributes greatly to the constant improvement on the quality of research papers published in the Chinese Journal of Laboratory Medicine. Compared with the status of international laboratory medicine and the level of international journals of laboratory medicine, there are still plant of rooms for improvement for the Chinese Journal of Laboratory Medicine, and arduous efforts are needed to further improve the quality of research papers published in the Chinese Journal of Laboratory Medicine.
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Objective:To explore the risk stratification and prognostic significance of loss of chromosome Y (LOY) in patients with multiple myeloma (MM).Methods:The clinical data of 193 male patients with newly diagnosed MM admitted to Zhongshan Hospital of Fudan University from January 2018 to January 2020 were analyzed retrospectively and divided into a normal karyotype group(178) and a LOY karyotype group (15) according to the results of their primary conventional cytogenetics. Rank sum test, 2×2 chi-square test and independent sample t-test were used to compare laboratory findings, such as liver and kidney function, immunohistochemistry and cytogenetics, treatment efficacy and survival prognosis, between the two groups. The clinical prognostic significance of LOY was summarized through survival analysis and Cox regression. Results:Among the newly diagnosed male MM patients, 8%(15/178) were confirmed with LOY cases. The proportion of patients with Revised International Staging System(R-ISS) stage Ⅲ was significantly higher in the LOY group (8/15) than that in the normal karyotype group (40/178)(χ 2=7.052, P<0.01). A higher proportion of 1q21 amplification also occurred in the LOY group (10/13 vs 77/162)(χ 2=4.159, P<0.05). The proportion of complete response(CR)/stringent complete response(sCR) in the normal karyotype group after the fourth chemotherapy (63/171) was significantly higher than that in the LOY group (1/15)(χ 2=5.564, P<0.05). The proportion of progressive disease (PD) was lower in the normal karyotype group (16/171 vs 4/15) (χ 2=4.306, P<0.05). The 2-year progression-free survival (PFS) of MM patients for the LOY group was significantly shorter compared to that for the normal karyotype group ( Z=?3.201, P<0.01). Univariate survival analysis showed that PFS was significantly shorter in newly diagnosed MM patients with Creatinine(Cr)≥93 μmol/L, β 2-microglobulin (β 2-MG)≥4.0 mg/L, serum free light chain(sFLC)<0.06, bone marrow plasma cells (BMPC)≥30%, R-ISS stage Ⅲ, failure to achieve CR/sCR after the fourth chemotherapy, with LOY, 1q21 amplification, P53 deletion and t(4;14) ( P<0.05). Cox regression analysis showed that Cr≥93 μmol/L( HR=4.460, 95% CI 1.615-12.314, P=0.004), sFLC<0.06( HR=2.873, 95% CI 1.206-6.849, P=0.017), failure to achieve CR/sCR after the fourth chemotherapy( HR=3.522, 95% CI 1.437-8.634, P=0.006)and with LOY( HR=3.485, 95% CI 1.473-8.249, P=0.006)were independent risk factors for PFS in newly diagnosed MM patients. Conclusions:LOY is an independent risk factor for poor prognosis. It is important for the clinical outcome and prognosis of patients with newly diagnosed MM, and may become a novel clinical assessment indicator.
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Cardiometabolic disease mainly refers to the cardiovascular disease caused by the increased metabolic burden, including hypertension, diabetes, dyslipidemia, coronary heart disease, stroke, and so on. In China, the incidence of this kind of disease is continuous rising, and cardiometabolic disease is more and more diagnosed in patients in younger age. Currently, continuous exploration of new biomarkers and detection methods, standardization of testing techniques, strengthening of clinical and laboratory communication and cooperation, and cultivation of interdisciplinary talents are the major focuses in term of promoting the development of laboratory medicine in the field of cardiovascular metabolism.
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We report a case of a 44-year-old female patient with acute liver failure and hepatic encephalopathy. The patient received artificial liver treatment and underwent allogeneic orthotopic liver transplantation at 14 days after admission. Laboratory examination reported a small number of green cytoplasmic inclusions in neutrophils on the peripheral blood smear at 68 days after admission. The patient eventually died of liver failure at 71 days after admission. Green inclusions are bright green or blue-green inclusions presented in the cytoplasm of neutrophils in Wright-Giemsa stained peripheral blood smears, and is associated with liver failure and high short-term mortality.
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Clinical mass spectrometry plays an increasingly important role in the analysis of hormones, drugs, vitamins, amino acids,etc. The analysis of large molecular peptides and proteins by mass spectrometry has also attracted wide attention. The development of clinical mass spectrometry technology still faces many challenges. With continuous efforts, clinical mass spectrometry technology will have a better future.
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Objective:This work aims to evaluate the clinical values of comprehensive genomic profiling examination based on circulating tumor DNA (ctDNA) in advanced lung cancer patients.Methods:This is a single-center, retrospective study that collected peripheral blood samples from patients with advanced lung cancer and performed gene mutation analysis using the TruSight Oncology 500 ctDNA assay kits. Between February 2022 and March 2023, a total of 82 patients were enrolled in Zhongshan Hospital, Fudan University, and 76 patients were included in the final analysis.According to the AMP/ASCO/CAP guidelines, mutations of targeted genes were divided into four levels (Tier I-IV), and the effectiveness of targeted therapy guided by ctDNA was evaluated. Descriptive statistics were used for basic characteristics, and the analysis of factors related to tumor mutational burden (TMB) was performed using the rank-sum test.Results:The ctDNA detection success rate was 92.7%(76/82).The median turnaround time for ctDNA testing was 10.5 days (9,13 days). At least one actionable mutation (Tier I or Ⅱ) was detected in 82.9%(63/76) of patients, and 28.6% (18/63) of patients received matched therapy, achieving a disease control rate of 18/18 and an objective response rate of 12/18.Conclusion:Comprehensive genomic profiling based on ctDNA can effectively identify actionable alterations in patients with advanced lung cancer and provide valuable information for matched therapy.
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Objective:To monitor nucleic acid contamination and evaluate the biosecurity risk in a temporary 2019-nCoV nucleic acid testing laboratory situated within designated infectious disease care facilities.Methods:Quantitative real-time PCR technology was used to detect nucleic acid contamination in samples collected from high-risk experimental activity areas and the surface of the laboratory staff′s personal protective equipment. Sampling was conducted every Monday from March 14 to May 16, 2022, both during and after disinfection procedures.Results:A total of 760 samples were collected from 40 sampling sites. A total of 27 out of 100 samples (27%) collected from 8 sampling sites in the sample processing area were positive. Among them, the contaminated area of biological safety cabinet, the outer surface of the sample transport box, and the sample rack were found to have the highest positive detection rates, with rates of 5/10, 4/10, and 6/10, respectively. Ten out of 140 samples (7.1%) obtained from 7 sampling sites in the nucleic acid detection area showed positive results. The inner wall of sample transfer window and the door handle of the nucleic acid detection area had the highest positive detection rates, both at 4/20. The Ct values for the target genes from the positive samples in the sample processing area were significantly higher than those from the nucleic acid detection area. The detection rate for nucleic acid contamination on the surface of the personal protective equipment of the laboratory staff was 20% (16/80), and the positive detection rate of the outer gloves from operator during the experiment reached 9/10. After disinfection, the nucleic acid residues on the surfaces of the various areas of the 2019-nCoV nucleic acid laboratory and the surfaces of the personal protective equipment of the laboratory staff were observed to be effectively removed.Conclusions:During experimental operation, the positive detection rate and nucleic acid contamination intensity of 2019-nCoV are higher in the sample processing area compared to those in the nucleic acid detection area. The laboratory staff are exposed to high biosecurity risk during the experiment. Implementing a scientific disinfection process can significantly reduce the risk of 2019-nCoV residues from the laboratory environment and the surface of the personal protective equipment of the laboratory staff, ensuring the accuracy of inspections and the safety of the laboratory staff.
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Objective:To assess the role of Delta-RBC parameters in the automated hematocrit analyzer RET channel for the recognition of cold agglutinins (CA) and to explore the value of RET channel optical method parameters in correcting interference with CA.Methods:This is a retrospective study. The Cas group included 68 samples with Cas interference and the control group included 45 samples without CA interference. All specimens were collected from Zhongshan Hospital Fudan University Outpatient Department from January 2022 to January 2023. Specimens in both the CA and Control group were examined using the CBC+RET channel at room temperature. Recorded and calculated the Impedance method test parameters RBC-I, HGB, HCT-I, MCH-I, MCV-I, MCHC-I and the Optical method test parameters RBC-O, HCT-O, MCH-O, R-MFV, MCHC-O, Delta-RBC. Examined the specimens in the CA group using the CBC channel after prewarmed at 37 ℃ for 2 h, and Impedance method parameters RBC-I 37 ℃ 2 h, HGB 37 ℃ 2 h, HCT-I 37 ℃ 2 h, MCH-I 37 ℃ 2 h, MCV-I 37 ℃ 2 h, MCHC-I 37 ℃ 2 h were recorded. The ROC curves were used to analyze the discrimination efficacy of Delta-RBC in identifying CA interference, and the Bland-Altman method was used to analyze the consistency between the results of the optical method RBC parameters at room temperature and the results of the impedance method after prewarming. The correlation analysis was performed using Pearson and Spearman correlation analysis to analyze the results of the RBC parameters before and after prewarming in the CA group. Result:If Delta-RBC was used as diagnostic indicators for the identification of CA interference, the best cut-off value was 1.065, with AUCs of 0.998. In the CA group, the correlation coefficients between RBC-O, HCT-O, R-MFV, MCH-O, MCHC-O, and RBC-I 37 ℃ 2 h, HCT-I 37 ℃ 2 h, MCV-I 37 ℃ 2 h, MCH-I 37 ℃ 2 h, MCHC-I 37 ℃ 2 h were 0.985, 0.981, 0.729, 0.870, and 0.649, respectively. The percentages within the limits of agreement of the percentage differences between the results of the two methods were 95.6%, 92.6%, 95.6%, 94.1%, and 95.6%, respectively. Conclusions:The RBC parameter Delta-RBC from RET channel optical method can be used as an indicator to effectively assist in the clinical determination of the presence of CA. Reporting results using the optical method RBC parameters of the RET channel can correct the interference of CA without specimen pre-treatment and obtain more correct results of complete blood counts.
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Objective:To compare the detection performance of serum free light chain (sFLC) in two platforms and evaluate the comparability of serum free light chain results in patients with multiple myeloma (MM).Methods:To evaluate the detection performance (repeatability, accuracy, linear range, reference range, interfering substances, etc.) of sFLC kit based on polyclonal antibodies. Spearman correlation analysis and Bland-Altman were used to analyze 214 sFLC results obtained on two detection platforms at the same time to evaluate the correlation between the results of the two methods and analyze the causes of methodological bias. 119 cases with aMM and 23 cases of disease control group (AL, WM, POEMS syndrome, MGUS, diffuse large B-cell lymphoma) initially diagnosed in the hematology department of Zhongshan Hospital of Fudan University from March 2020 to March 2021 were all included. A retrospective analysis was conducted to calculate the area under the curve of receiver operating characteristic (AUC-ROC) and obtain the optimal sensitivity and specificity cut-off points for the diagnosis of MM patients on monoclonal antibody platform.Results:Repeatability, accuracy, linear range, reference interval and anti-interfering capacity of the detection platform based on polyclonal antibodies were verified to meet clinical needs. The overall consistency of FLC/κ, FLC/λ and κ/λ ratios in two methods was 89.3%, 84.1% and 77.1% respectively; but the correlation results were highly heterogeneous. The correlation coefficient of FLC/κ R 2 was 0.922( P<0.001), while the correlation coefficients R 2of the FLC/λ and κ/λ ratios were only 0.349 and 0.441( P<0.001). After segment analysis, it was found that the correlation of FLC/λ was improved within the linear range and R 2 could rise to 0.78( P<0.001). Compared with monoclonal antibody platform, the vast majority points of FLC/κ fell within the 95% limit by Bland Altman analysis. While the results of FLC/λ on polyclonal antibody platform showed significant positive bias. The AUC of MM diagnosis on monoclonal antibody platform was 0.751 ( P=0.001), and the optimal cutoff value was 24.67. Conclusion:The overall consistency between the two platforms was good, but there were significant differences between the results, so they were not comparable and could not be interchanged. For monitoring the prognosis of patients with multiple myeloma, the same platform should be selected for testing.
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The delta checks are one of the patient-based quality control options to identify the errors and the significant changes in patients′ condition. Compared with the traditional internal quality control method, the delta checks have the characteristics of real-time monitoring, with no additional detecting cost, thus the delta checks are widely used in clinical laboratories. In addition, the delta checks are also useful in the auto-verification system to screen out the abnormal results for manual verification. This article reviewed the delta checks′ development history, parameters selection, application values in quality control and auto-verification.
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Dyslipidemias played the critical role in cardiovascular disease risk assessment. Evaluate the benefits and limitations of advanced clinical lipids research and measurements would be important for assessing and managing cardiovascular risk.
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Point-of-care testing (POCT) systems has been widely used in clinical practice. It is necessary to carefully review and summarize issues related to the testing speed, cost, and quality of POCT. Guidelines and standardization documents for clinical application of POCT need to be formulated as soon as possible. These works would contribute to the accuracy and reliability of POCT results and help the accurate application of POCT in clinical practice.
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Objective:To verify the performance of the next-generation sequencing (NGS) platform and evaluate the application of NGS, droplet digital PCR (ddPCR) and super amplification refractory mutation system (super-ARMS) in the detection of circulating free DNA (cfDNA) mutations in patients with non-small-cell lung cancer (NSCLC).Methods:A total of 75 patients with NSCLC in the respiratory department of Zhongshan Hospital Affiliated to Fudan University were enrolled. The standards, cfDNA from 25 patients with newly diagnosed and untreated NSCLC, and self-made mixed samples mixed with hemoglobin (1 000 mg /dl), bilirubin (500 mg/l), fat emulsion (2%), enterococcus gDNA and Escherichia coli gDNA were used to verify the blank limit, analytical sensitivity, precision, accuracy and specificity of NGS platform. The cfDNA mutations of 75 NSCLC patients were detected by ddPCR and NGS, and the mutation positive rates of the two platforms were compared. The linear relationship between the two platforms was compared by Pearson correlation test. 12 patients were selected by simple random sampling for the detection of plasma super-ARMS platform. The performance of three platforms in the detection of plasma cfDNA mutation in patients with NSCLC was compared.Results:The blank limit of NGS platform was set to 0.00%, the analytical sensitivity was 0.2%, the intra-assay precision and inter-assay precision were 100%. The test results were not affected by endogenous hemoglobin, bilirubin or fat emulsion in plasma or exogenous DNA interference, and the analysis specificity was good. The mutation positive rates of plasma cfDNA in 75 NSCLC patients detected by ddPCR and NGS were 61.33% and 60.00%, respectively. The complete coincidence rate was 89.33%, which suggests there was a positive correlation between the mutation abundance of NGS and ddPCR ( r=0.984, P=0.001). Among the plasma of 12 NSCLC patients, the results of NGS, ddPCR and super-ARMS were completely consistent in 7 cases, including 2 wild-types and 5 mutants. Conclusion:The NGS platform was verified to be useful for cfDNA mutation detection in patients with NSCLC. The ddPCR, NGS and super-ARMS have their own advantages in detecting cfDNA mutations in patients with NSCLC.
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Taking advantage of current advancements of the laboratory information system, patient-based real-time quality control (PBRTQC) has become a hot research topic with the potential of significantly improving the quality of clinical laboratories. PBRTQC has several advantages over conventional internal quality control, including the merit of continuous error detection, low maintenance cost, and no fear of loss of quality control due to commutability issue. This review summarized the recent research progress in PBRTQC. The review described the fundamental theories of PBRTQC and explained how to implement PBRTQC in clinical laboratories in detail. The review also provided prospective view on how to solve the current problems and what are the future research directions of PBRTQC.
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Objective:To investigate the expression of four cancer stem cell (CSC) markers (EpCAM, CD133, CD90 and CD24) in hepatocellular carcinoma tissues and peripheral blood circulating tumor cells (CTC),their value in the prognosis of patients with hepatocellular carcinoma.Methods:A total of 50 hepatocellular carcinoma tissues and 29 peripheral blood sample from 50 patients with hepatocellular cancer treated in Zhongshan Hospital Fudan University from October 2013 to September 2014 were collected and analyzed by flow cytometry or qRT-PCR to examine the expression of EpCAM, CD133, CD90 and CD24. The clinical data of patients were collected, including tumor size, tumor number, satellite lesions, vascular invasion, Edmondson stage, BCLC stage and liver cirrhosis, etc. The correlation between the expression of four markers in hepatocellular carcinoma tissues and CTC with the clinical data and survival time of patients were compared.Results:The positive expression rates of EpCAM, CD133, CD90 and CD24 in hepatocellular carcinoma tissues were 66% (33/50), 18% (9/50), 60% (30/50) and 56% (28/50); the positive expression rates in CTC were 55% (16/29), 38% (11/29), 31% (9/29) and 59% (17/29). CD90 expression in hepatocellular carcinoma tissue was positively correlated with the occurrence HCC liver cirrhosis ( P<0.05), while CD133 expression was negatively correlated with the 5-year survival rate of patients ( P<0.05). The expression of EpCAM and CD24 in peripheral blood CTC were closely related to the patient′s Edmondson stage ( P<0.05). The survival time of patients with CD133 positive expression in hepatocellular carcinoma tissue was lower than those without CD133 expression ( P<0.05); the survival rate of patients with EpCAM expressed in either tissue or peripheral blood CTC was lower than that of patients with EpCAM double negative expression ( P<0.05). The survival rate of patients with CD90 negative in HCC tissue and positive in peripheral blood was lower than that in patients with double negative/double positive in tissue and peripheral blood or patients positive in hepatocellular carcinoma tissue and negative in peripheral blood ( P<0.01). Conclusion:Different expression characteristics of four markers in cancer tissues and peripheral blood CTC might provide useful information about predicting prognosis of hepatocellular carcinoma. The expression of CD133 in tissues can be used as an important survival predictor of hepatocellular carcinoma patients. The differential expression of cancer markers in tissue samples and blood samples can provide more clinical prognostic information.