Table of Content

30 January 2015, Volume 30 Issue 1

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Introduction of the ICSH guidelines for the evaluation of blood cell analyzers, 2014 edition (Ⅱ)

HU Xiaobo, SONG Ying, WANG Qing, JIN Daming, XIONG Lifan

2015, 30(1):  1-6.  DOI: 10.3969/j.issn.1673-8640.2015.01.001

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Introduces the International Council for Standardization in Haematology (ICSH) guideline for the evaluation of blood cell analyzers, 2014 edition. In this guideline, the performance evaluation, including sampling mode, within-run precision, between-run precision, carry-over, linearity, sample stability, reference interval, accuracy and comparability of haematology analyzers, should be conducted through validation and verification by manufacturers. The assessment of flow cytometry-based or digital imaging-based leukocyte differential count, nucleated red blood cell count and immature granulocyte count is recommended by flow cytometry immune-phenotypic counting as a reference method. A part of validation or transference of instrument should be performed by each clinical laboratory before reporting patient results.

Quality management of clinical laboratories in Taiwan

JU Yi, ZHONG Jianming, ZHU Yuqing, XIAO Yanqun, WANG Qing.

2015, 30(1):  7-9.  DOI: 10.3969/j.issn.1673-8640.2015.01.002

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Through visiting 4 clinical laboratories and 1 proficiency testing organization in Taiwan, we have a better understanding of Taiwan quality management methods, which can help as with improving our quality management and promoting laboratory accreditation. Taiwan quality management can be classified into 3 models. The first is seamless management model based on information system. The quality management requirements are fitted into each working procedure by information integration function. This has the advantage of practice and substantially improves efficiency. The second is to manage the duties of important positions. To achieve the participation of quality management, the passive work is changed into positive work by competing position provision and by stepwise control. The third is performance appraisal based on government management and control and orderly market. This model makes use of human potential encouraging mechanism, and effectively promotes the development of laboratories. Clinical laboratories in Taiwan improves their quality management by scientific methods and human-centered views. Laboratory staffs are encouraged to actively get involved in the establishment of quality management system, which improves the test quality and meets the clinical requirements.

Analysis on the antibiotic resistance of Klebsiella pneumoniae isolated from neurosurgical intensive care unit

WANG Bei, AI Fuqi, ZOU Xue, JIANG Xiaofei.

2015, 30(1):  10-12.  DOI: 10.3969/j.issn.1673-8640.2015.01.003

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Objective To analyze the antibiotic resistance and characteristic of Klebsiella pneumoniae isolated from neurosurgical intensive care unit in Huashan Hospital. Methods A total of 233 Klebsiella pneumoniae isolates were collected from neurosurgical intensive care unit. Antibiotic susceptibility of the isolates was determined. Results A total of 77.3% of 233 isolates of Klebsiella pneumoniae were collected from respiratory tract samples. The total resistance rates to antibiotics increased year after year. The resistance rates to piperacillin-tazobactam, cefoperazone-sulbactam, imipenem, meropenem and ertapenem for respiratory tract samples were obviously lower than those for non-respiratory tract samples (P<0.05). Conclusions There is a contrast between the antibiotic susceptibility of the respitatory specimens and the non respiratory specimens. The monitoring of nosocomial infection should be strengthened to avoid misusing antibiotics.

Drug resistance analysis on the drug resistance of Streptococcus agalactiae isolated from female urogenital tract

GAO Jing, LIU Xiaoyan.

2015, 30(1):  13-16.  DOI: 10.3969/j.issn.1673-8640.2015.01.004

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Objective To analyze retrospectively the drug resistance of Streptococcus agalactiae isolated from female urogenital tract, to understand the drug resistance tendency change of commonly-used antibiotics, and to guide the rational use of antibiotics in clinical application. Methods The routine culture was performed for female urogenital tract specimens collected from January 2009 to December 2012, and the identification and drug sensitivity test were performed for Streptococcus agalactiae by VITEK32 automatic bacterium identification analyzer. The D-inhibition zone trial was tested for isolates which were drug-resistant to erythromycin and sensitive or intermediary to clindamycin. Results There was a statistical significance in age distribution among 569 specimens from Department of Gynecology and 192 specimens from Department of Obstetrics (P<0.01). Drug resistance rate was 0 to vancomycin and <3% to penicillin and linezolid. The drug resistance rate of nitrofurantoin was <0.5% from 2009 to 2011 and suddenly increased to 7.2% in 2012. The drug resistance rate of tetracycline exceeded 90%. Erythromycin, clindamycin and levofloxacin reflected high drug resistance rates and such an increasing tendency year by year. The positive rate of D-inhibition zone trial was 21.7%. Conclusions It is required to rationally use antibiotics in clinical application according to the drug sensitivity test results. Meanwhile, the drug resistance monitoring in clinical application should be strengthened.

The clinical application of PCT in the diagnosis of systemic and local bacterial infections

MU Yuejing, WANG Weijia, YAN Haizhong, XU Quanzhong, KAN Lijuan, OUYANG Nengliang, HUANG Yanhua, ZHANG Xiuming.

2015, 30(1):  17-20.  DOI: 10.3969/j.issn.1673-8640.2015.01.005

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Objective To evaluate the clinical application significance of serum procalcitonin(PCT)quantitation determination in the diagnosis of systemic and local bacterial infections. Methods The Clinical and Laboratory Standards Institute (CLSI) EP15-A2 document was employed to verify the precision and accuracy of PCT determination by Roche Cobas E601 chemiluminescence analyzer (Cobas E601) and ensure instrument conditions being complied with the requirement of experiment. A total of 697 patients with systemic and local bacterial infections were enrolled. The results of serum PCT determination, blood culture, sputum culture, middle-urine culture and cerebrospinal culture were used to compare the change trends of serum PCT for systemic and local bacterial infections and for different-part infections. The difference of PCT in Gram-positive cocci and Gram-negative bacilli infections was analyzed. Results Cobas E601 had good repeatability, and the total coefficient of variation (CV) was <3.59%. The fixed calibration bias was <5.0% and met the clinical determination requirements. For serum PCT, there was significance for systemic and local bacterial infections with having increasing serum PCT level(P<0.05). For local bacterial infection, there was no statistical significance for respiratory system, urinary system and nervous system infections (P>0.05). For systemic bacterial infection, there was higher serum PCT level for patients with Gram-negative bacilli infection than with Gram-positive cocci infection (P<0.05). For local bacterial infection, there was no statistical significance for patients with Gram-negative bacilli infection and Gram-positive cocci infection (P>0.05). Conclusions In patients with systemic and local bacterial infections, serum PCT level has high expression, and serum PCT level can not identify local bacterial infection Gram-negative bacilli and Gram-positive cocci infections.

Analysis on the missing detection of CD5 in chronic lymphocytic leukemia and its countermeasures

TANG Gusheng, LIU Min, HU Xiaoxia, GAO Lei, YANG Jianmin, WANG Jianmin.

2015, 30(1):  21-25.  DOI: 10.3969/j.issn.1673-8640.2015.01.006

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Objective To analyze the variation of CD5 caused by different fluorescein-labeled antibodies on B cells in patients with chronic lymphocytic leukemia(CLL) and the influence on clinical diagnosis. Methods Flow cytometry and SYSMEX XE2100 automated hematology analyzer were used for the detection of immunophenotype and blood cell counts of 3 CLL patients with different expression intensities of CD5, respectively. 1 patient with B-cell acute lymphocytic leukemia(B-ALL) was selected as negative control. Result In 4 patients with abnormal B lymphocytes, using fluorescein isothiocyanate(FITC) fluorescein-labeled antibody, no independent groups expressing CD5 were identified, and the proportions of CD5 positive cells were 47.1%, 17.7%, 6.7% and 7.9%. For ≥20% as standard, there was only 1 positive case. Re-tested with PE-Cy7 fluorescein-labeled antibody, however, there were independent groups of CD5 positive cells in case 1 and 2, and there was still a continuous expression pattern of CD5 in case 3. The proportions of CD5 positive cells increased to 91.1%, 70.3% and 38.2% in these 3 specimens from CLL patients. In case 4, the specimen from B-ALL patient, the proportions of CD5 positive cells were negative in the 2 conditions. Case 1, 2 and 3 were typical CLL phenotyping. Conclusions CD5 is often abnormally expressed on B cells in CLL patients, but its expression intensity is significantly lower than that on T cells. Using weak fluorescein-labeled antibodies might miss CD5 expression on B cells, and thus it will affect the clinical diagnosis because of leak detection. It should select appropriate fluorescein-labeled antibodies for weakly expressed antigens and conduct validation and comparison to determine its accuracy, which is an important safeguard for the provision of correct disease phenotype.

Significance of comprehensive analysis on the diagnosis of myeloproliferative disease

ZHOU Ping, TANG Jibin, JIAO Ruibao, ZHANG Wen, CAO Chunluan, CHONG Huifeng, ZHONG Weijun

2015, 30(1):  26-30.  DOI: 10.3969/j.issn.1673-8640.2015.01.007

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Objective To investigate the significance of V617F mutation in JAK2 gene combining cytometry and bone marrow cell morphology on the diagnosis of myeloproliferative disease (MPD). Methods Of the 102 cases whose peripheral blood number increased with various degrees, the real-time fluorescence quantitation polymerase chain reaction (PCR) was applied to detect the V617F mutation in JAK2 gene, and the cell morphology of bone marrow was observed. Results In the 54 cases of MPD, including polycythemia vera (PV) and essential thrombocythemia (ET) and primary myelofibrosis (PMF), the positive rates of JAK2 V617F mutation were 91.7%, 50.0% and 66.7% respectively, while JAK2 V617F mutation was not detected in the 48 non-MPD cases with statistical significance (P<0.05). Between MPD and non-MPD groups, the value of part blood cell count had statistical significance (P<0.05). There existed a certain characteristic on bone marrow cell morphology in MPD group. Conclusions Compared with patients with secondary blood disease, MPD patients display higher frequency of V617F mutation in JAK2 gene, and there is vital significance for the diagnosis with combination of cytometry and bone marrow cell morphology.

Levels of CD4⁺CD25^highFoxP3⁺ regulatory T lymphocytes in peripheral blood and plasma IFN-γ, IL-10 of patients with lymph tuberculosis and their clinical significance

ZHANG Guoying, NIU Xiaohong, XU Weiping, XIA Xia.

2015, 30(1):  31-35.  DOI: 10.3969/j.issn.1673-8640.2015.01.008

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Objective To study the changes of peripheral blood CD4⁺CD25^highFoxP3⁺ regulatory T lymphocytes (Treg) and interferon gamma (IFN-γ), interleukin 10 (IL-10) levels of patients with lymph tuberculosis and their clinical significance. Methods The levels of peripheral blood CD4⁺CD25^highFoxP3⁺ Treg, IFN- γ and IL-10 of 100 lymph tuberculosis patients and 30 healthy subjects were determined by flow cytometry. Results Compared with the healthy controls, the levels of peripheral blood CD4⁺CD25^highFoxP3⁺ Treg and IL-10 levels in lymph tuberculosis group increased with statistical significance (P<0.05), while IFN-γ level decreased (P<0.05). Among the caseous, proliferation and mixed types of lymph tuberculosis, there was no significant difference (P>0.05). Conclusions Cellular immune function is significantly abnormal in patients with lymph tuberculosis. CD4⁺CD25^highFoxP3⁺ Treg, IFN- γ and IL-10 play important roles in the onset process of lymph tuberculosis.

Observation and analysis on lymphocyte subsets of peripheral blood in oral mucosa diseases

WU Jingjing, LU Yibin, HUANG Qijie, YAO Yuting, GU Wenli.

2015, 30(1):  36-39.  DOI: 10.3969/j.issn.1673-8640.2015.01.009

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Objective To observe the changes of lymphocyte subsets of peripheral blood in 2 common oral mucosa diseases, recurrent aphthous ulcer (RAU) and oral lichen planus (OLP), and to investigate their immunological pathogenesis. Methods By flow cytometry, the lymphocyte subsets (total T cells, CD4⁺ T cells, CD8⁺T cells, CD4⁺/CD8, NK cells and B cells) in the peripheral blood of patients with RAU (105 cases), OLP (70 cases) and healthy controls (65 cases). The difference was analyzed comparatively between the groups. Results Compared with healthy controls, total T cells and CD8⁺T cells in RAU group increased (P<0.05, P<0.01), but CD4⁺T cells, CD4⁺/CD8⁺ and B cells decreased in RAU group (P<0.01, P<0.001, P<0.05). In addition, CD8⁺T cells in OLP group were lower than those in control group (P<0.05). The total T cells and CD8⁺T cells in RAU group were higher than those in OLP group (P<0.01, P<0.001), while CD4⁺T cells, CD4⁺/CD8⁺ and NK cells in RAU group were significantly lower (P<0.01, P<0.001, P<0.05). Conclusions Owing to the imbalanced lymphocyte subset percentages, the cellular immune function of patients with RAU or OLP is often found to be abnormal, but their immune states are not completely same. So, the application of selective immunologic intervention may be the promising therapeutic measure to those oral mucosa diseases.

Research on the relationships of Lp-PLA2 and Hcy levels with atherosclerotic cardio-cerebrovascular disease

CHENG Qing

2015, 30(1):  40-43.  DOI: 10.3969/j.issn.1673-8640.2015.01.010

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Objective To investigate the relationships of plasma lipoprotein-associated phospholipase A2 (Lp-PLA2) and homocysteine (Hcy) with atherosclerotic cardio-cerebrovascular disease. Methods A total of 198 patients with atherosclerotic cardio-cerebrovascular disease were classified into the group with traditional risk factors and the group without traditional risk factors. A total of 58 healthy subjects were enrolled as healthy control group. The plasma concentration of Lp-PLA2 was detected by double-antibody sandwich enzyme-linked immunosorbent assay(ELISA), and Hcy level was detected by enzymatic cycling method. Results Compared with healthy control group, Lp-PLA2 and Hcy levels increased significantly in both groups with and without traditional risk factors (q=44.236, P=0.000; q=11.403, P=0.000; q=50.025, P=0.000; q=15.777, P=0.000). No statistical significance was found between groups with and without traditional risk factors for Lp-PLA2 and Hcy levels (q=2.299, P=0.083; q=2.586, P=0.069). Lp-PLA2 and Hcy levels in groups with and without traditional risk factors had no correlation (r=0.069, P=0.375;r=0.053, P=0.566). Balanced other risk factors, Lp-PLA2 and Hcy levels were independently correlated with coronary heart disease (OR=1.917, P=0.005; OR=1.883, P=0.013). Higher positive rate can be got by Lp-PLA2 and Hcy combined detection than the individual detections in both groups with and without traditional risk factors (χ²=8.124, P=0.004; χ²=3.933, P=0.045). Conclusions Plasma Lp-PLA2 and Hcy levels can be used in risk assessment of atherosclerotic disease,and the combined detection has some predictive significance for atherosclerotic cardio-cerebrovascular disease.

Verification on the result accuracy of ALP by two routine measurement systems according to IFCC reference method

WU Lishan, CHEN Shufen, HU Zhibao, SHEN Min, ZHOU Jun.

2015, 30(1):  44-48.  DOI: 10.3969/j.issn.1673-8640.2015.01.011

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Objective To evaluate the accuracy of serum alkaline phosphatase (ALP) determination by routine measurement systems. Methods The fresh serum samples were determined simultaneously by the International Federation and Clinical Chemistry(IFCC) reference method and 2 routine measurement systems (A method and B method). The accuracies were evaluated according to the guideline of EP9-A2, the Clinical and Laboratory Standards Institute(CLSI). Meanwhile, the consistency of results between routine measurement systems and IFCC reference method was evaluated by improved Bland-Altman graphical analysis. The accuracies were evaluated comprehensively. The matrix effects of A and B methods were evaluated according to CLSI EP14-A2. Results The regression equation of A method with IFCC reference method was Y_A = 0.982 9X_IFCC+0.010 8, and the average bias was -1.1%. The regression equation of B method with IFCC reference method was Y_B=0.938 3X_IFCC + 0.012 9, and the average bias was -5.5%. The correlation equation between A method and B method was Y_B=0.955 2X_A+0.001 36, R²=0.997 6. The calibrator matrix effects of both A method and B method were found, and the calibrator matrix effect of A method was more significant. Conclusions The accuracy of A method is consistent with that of IFCC reference method, and the accuracy of B method is inconsistent with that of IFCC reference method.

Application evaluation of latex-enhanced immunonephelometry method to determine serum amyloid A

JIANG Jianwei, YANG Yu, YING Chunmei

2015, 30(1):  49-52.  DOI: 10.3969/j.issn.1673-8640.2015.01.012

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Objective To establish the analysis performance of serum amyloid A (SAA) detected by latex-enhanced immunonephelometry method with point-of-care test (POCT), and to evaluate precision, linearity, accuracy and so on. Methods According to relevant documents of the Clinical and Laboratory Standards Institute (CLSI), the methodological evaluation was performed. Results The sensitivity of this method was 3.52 mg/L. The within-run and inter-day coefficients of variation (CV) of this method were <8% and <10%, respectively, with strong anti-interference ability. When the level of hemoglobin was ≤4.0 g/L, the level of bilirubin was ≤400 μmol/L, the level of rheumatoid factor was ≤1 621 U/L, and the level of triglyceride was ≤10 mmol/L, there was no influence on the results. There was a good correlation with that of imported N Latex SAA assay (Y=1.052 1X+0.001 5, r=0.998 3). The detection linear range of this method was 5-200 mg/L. Conclusions The latex-enhanced immunonephelometry method for SAA is convenient, and it can be finished within 3 min. Its performance meets the requirements for in vitro diagnostic reagents, and it is suitable for the detection of serum samples in clinical use.

Epidemiology and resistance mechanism study of carbapenem-resistant Acinetobacter baumannii

ZHANG Guanghui, CAO Xueping, YU Jing, LIU Ying, GE Yan

2015, 30(1):  53-57.  DOI: 10.3969/j.issn.1673-8640.2015.01.013

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Objective To understand the drug resistance mechanisms and epidemiology of carbapenem-resistant Acinetobacter baumannii in order to guide the rational use of antibiotics and to provide the reference for the control of nosocomial infections. Methods A total of 116 Acinetobacter baumannii (110 carbapenem-resistant isolates and 6 carbapenem-sensitive isolates) isolated from clinical specimens were classified and analyzed according to endemic areas and specimen types. The isolates were analyzed by repetitive-sequence polymerase chain reaction (REP-PCR) for genotyping. Beta-lactamase genes (OXA, IMP, VIM and soon) and efflux pump-encoding genes (adeA, adeB, adeC, adeJ, adeG) were amplified by PCR. Real-time fluorescence quantitation PCR was performed to compare the expression level of the efflux pump-encoding genes in resistant and sensitive isolates. Results The isolation rate of resistant isolates in intensive care unit (ICU) was the highest-level, and the isolates were mainly isolated from sputum. The genotype types of carbapenem-resistant and -sensitive Acinetobacter baumannii were classified into 7 categories by REP-PCR. All the carbapenem-resistant isolates were type A. OXA-23 and OXA-51 were positive in 110 carbapenem-resistant isolates. However, OXA-24, OXA-58, IMP-1, IMP-2, VIM-1 and VIM-2 could not be detected in all the isolates. The positive rates of the efflux pump-encoding genes (adeA, adeB, adeC, adeJ and adeG) were 100%. The expression level of efflux pump-encoding genes (adeA, adeB, adeC) in resistant isolates was significantly higher than that in sensitive isolates. Conclusions The main popular type of carbapenem-resistant Acinetobacter baumannii is type A. Clinical staff should pay attention to this phenomenon. The expression of OXA-23 and OXA-51 overexpression of adeA, adeB and adeC efflux pump-encoding gene may play an important role in carbapenem-resistant Acinetobacter baumannii.

Establishment on a laboratory physician-centred communication system

MA Xiaowei, QIN Juanxiu, HE Lei, LI Min.

2015, 30(1):  64-67.  DOI: 10.3969/j.issn.1673-8640.2015.01.016

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In order to adapt to the rapid development of laboratory medicine, laboratory physician was set up in specialist classification to promote the communication of laboratory and clinic by the Ministry of Health and Chinese Medical Doctor Association in October, 2003. On the current medical circumstances, however, the duty of laboratory physician is not clear in most hospitals about 10 years later. The purpose of the article is to establish a laboratory physician-centred communication system to provide advice and suggestions for laboratory physicians.

Research on the present stage and development of continuous medical education for clinical laboratory in Shanghai

WANG Yin, WANG Qingzhong.

2015, 30(1):  68-71.  DOI: 10.3969/j.issn.1673-8640.2015.01.017

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Continuous medical education is a main way that medical technical staffs get new methods and obtain new knowledge, and it is a method for the maintain and supplement of professional education. In this paper, through understanding the present stage of continuous medical education, the existing problems at the present stage of continuous medical education are found and analyzed, and the corresponding countermeasures and improvement measures are put forward, which will increase the levels of clinical laboratory medical technical staffs with important significance in Shanghai.

Research progress on platelet function testing and its application in a variety of diseases

CAI Yuchan, LI Zhi.

2015, 30(1):  72-75.  DOI: 10.3969/j.issn.1673-8640.2015.01.018

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Abnormal platelet quantity and quality are important factors leading to bleeding and thrombotic diseases. Meanwhile, they are also involved in the pathological process of some diseases, such as diabetic complications, angiocardiopathy, tumor and inflammatory. Platelet function testing, particularly for aggregation function and activation function of platelets, can provide a approach for the prevention and diagnosis of clinical diseases, especially for the treatment and monitoring of diseases.

The research progress of gold nanoparticle in clinical laboratory diagnosis

LI Huimin, FU Qihua, WANG Jing

2015, 30(1):  76-79.  DOI: 10.3969/j.issn.1673-8640.2015.01.019

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Gold nanoparticle is a type of nanomaterials, which is widely used in nanodiagnosis, and exhibits high surface-to-volume ratio, unique physical properties, excellent biocompatibility and chemical stability. Gold nanoparticle may easily be functionalized by modification with special molecules. In this review, we mainly focus on the recent advances of gold nanoparticle research in clinical laboratory diagnosis.

The protective effect of neutrophil gelatinase-associated lipocalin in acute kidney injury

ZHANG Yali, CUI Liyan, ZHANG Jie.

2015, 30(1):  80-84.  DOI: 10.3969/j.issn.1673-8640.2015.01.020

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Acute kidney injury(AKI) is common in clinical practice, and the prognosis is so poor that needing early intervention, but now clinical diagnosis often depends on the functional indicators, such as serum creatinine (SCr) determination, however, due to the detection technology limitations, abnormal SCr appears much late, and the results may have some deviations, which is unfavorable about timely diagnosis and intervention of AKI, so it is urgent for a clear biomarker of early kidney damage. In recent years, scholars have been looking for the early biological markers of AKI, gradually finding some potentially useful indicators, such as neutrophil gelatinase-associated lipocalin(NGAL), kidney injury molecule-1(KIM-1), interleukin 18(IL-18), cystatin C(Cys C), N-acetyl-beta-D-glucosaminidase(NAG), liver fatty acid binding protein(L-FABP) and so on. NGAL as an early biomarker of AKI attracts much attention. Some studies show that NGAL also play an important role in protecting kidney inflammation and tumorigenesis, but the specific mechanism has not been fully understood yet. This review will discuss the protective role of NGAL in the occurrence of AKI and the related mechanisms.

Clinical application significance of serum total bile acid detection in pediatric liver diseases

LIU Jintao

2015, 30(1):  85-89.  DOI: 10.3969/j.issn.1673-8640.2014.06.021

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Serum total bile acid is a highly sensitive and specific indicator for the diagnosis and treatment of hepatobiliary disorders. It has been valued highly in clinic, especially in pediatric liver diseases. Serum total bile acid has important significances in the diagnosis, treatment efficacy, disease progression and prognosis. Serum total bile acid can be used as an important reference parameter in the clinical diagnosis and treatment of pediatric liver damage and liver-related diseases.