Laboratory Medicine

The comparative study on the testing results between 2 analyzers for determining C reactive protein

SONG Na;ZHANG Jiayun;YU Xiaohong;GUO Xia

2012, 27(4): 257-260.

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Objective To evaluate the analysis performance of testing results between speed scattering turbidimetric method and transmitted immunoturbidimetric method for determining C reactive protein(CRP). Methods The speed scattering turbidimetric method (Siemens Healthcare Diagnostica BN-Ⅱ specific protein analyzer) and transmitted immunoturbidimetric method (Finland Orion Diagnostica QuikRead CRP analyzer) were used to determine CRP, and the precision, linearity, interference, correlation and bias were evaluated methodologically. Results The total coefficient of variation (CV) of the BN-Ⅱ specific protein analyzer was <6%, when the concentration of CRP was 2.0-80.0 mg/L. The total CV of the QuikRead CRP analyzer was <7%, when the concentration of CRP was 8.0-80.0 mg/L. The linearity range was 8.0-70.0 mg/L. Haemoglobin (Hb) <10 g/L and bilirubin (Bil) <300 mg/L had no significant interference (<10%) for the assay. The interference (<10%) was not significant to the BN-Ⅱ specific protein analyzer when triglyceride (TG) <20 mmoL/L. When TG >15 mmol/L, the interference of low-value CRP samples was >10%, and there was no interference in high-value CRP samples for QuikRead CRP analyzer. The correlation analysis of 50 blood samples showed that both analyzers were correlated well (r=0.98，P<0.01). There was no significant bias from linearity regression through Cusum between the 2 analyzers（P>0.05）. The values of BN-Ⅱ specific protein analyzer were slightly lower than those of QuikRead CRP analyzer. Bland-Altman curve showed that the average bias of the 2 analyzers was -2.1. 〖WTHZ〗 Conclusions The precision, interference and linearity tests are suitable for routine CRP determination on the 2 analyzers. Although the results have bias, they meet the clinical requirements.

Application of different allowable total error in the performance evaluation of specific protein determination and selecting quality control rule

JIANG Lingli;XIAO Yanqun;WANG Qingzhong;ZHANG Jian

2012, 27(4): 261-265.

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Objective To evaluate the performance of specific protein under different allowable total error (TEa) of different international standards, apply 6 sigma (σ) metrics into selecting and improving quality control rule. Methods The data from internal quality control and the proficiency test of the National Center for Clinical Laboratory in 2009 and 2010 were collected respectively, and the imprecision and bias were calculated. TEa was quoted from Clinical Laboratory Improvement Amendments 88 (CLIA′88), German Guidelines for Quality and biological variation requirements. According to the coefficient of variation (CV), bias and TEa, σ was calculated, and quality control strategy was designed. In addition, the quality goal index (QGI) below 6σ was calculated to find problem and decide the priority improvement of precision and accuracy. Results According to IgA, IgG, IgM, C3, C4 and C reactive protein (CRP) with 3 different TEa in 2009, the σ were 0.53-6.95 respectively. The items were below 6σ except CRP, and under biological variation requirements the CRP σ was 6.95. In 2010, the σ of the 6 items was 1.33-29.39 respectively, and all items were above 6σ except IgG and C3 under biological variation requirements. Conclusions Using the σ metric, the performance and appropriate internal quality control can be chosen and developed economically and reasonably.

A study on the resistance mechanism of Morganella morganii to carbapenems

2012, 27(4): 266-270.

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Objective To investigate resistance mechanisms to carbapenems and transmission modes of 3 Morganella morganii isolates (M1， M2 and M3) isolated from patients in surgical intensive care unit (SICU). Methods The susceptibility of antimicrobial agents was detected by agar dilution method. The molecular epidemiological characteristics of the isolates were analyzed by enterobacterial repetitive intergenic consensus (ERIC)-polymerase chain reaction (PCR). The encoding genes of beta-lactamases were analyzed by specific PCR amplification. The transitivity and homology of plasmids were detected by plasmid conjugation and restriction endonuclease analysis. Genetic environments around bla_KPC were analyzed by sequencing. The changes of outer membrane permeability were analyzed by the electrophoresis of outer membrane protein. Results The bla_KPC-2 was detected by PCR in all 3 isolates and their transconjugants. M1 and M2 were the same clonal type. Carbapenem-resistance was successfully transfered by conjugation experiments. The bla_KPC-2 was located on dissimilar plasmids with quite different restriction profiles. The genetic environments around bla_KPC-2 were the same. The 3 original Morganella morganii isolates showed a loss of 38 000 outer membrane proteins, and additional 36 000 outer membrane proteins appeared compared with the Morganella morganii isolates susceptible to carbapenems. Conclusions The bla_KPC-2is detected in the 3 Morganella morganii isolates belonging to 2 clonal types . This gene encoded by dissimilar plasmids is likely located on the same mobile genetic elements by which bla_KPC-2 is possibly transferred between different plasmids. The KPC-2-producing and lack of outer membrane proteins together cause these 3 Morganella morganii isolates resistant to carbapenems possibly.

Study on the diagnosis of catheter-related bloodstream infection by retaining intravascular catheter methods

2012, 27(4): 271-274.

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Objective To study the diagnosis of catheter-related bloodstream infection (CRBSI) by retaining intravascular catheter methods, and to provide a method to decrease the unnecessary removing of the intravascular catheter for clinician. Methods According to the patient′s clinical symptoms, the clinician in the Central Intensive Care Unit of Huashan Hospital classified the patients into retaining and removeing the catheter for diagnosing the CRBSI from November in 2009. In the case of retaining the catheter, a set of blood (a bottle of aerobic blood culture and a bottle of anaerobic blood culture) was taken from peripheral vein and central venous catheter to perform blood culture. In the case of removing the catheter, the another 3 mL blood sample was collected and performed quantitative blood culture, and 2 sets of blood were send to determination. Results The 27 of 50 episodes of retaining catheter specimens were detected the growth of bacteria. The 10 of them were CRBSI by laboratory evidence plus clinical diagnosis. The 8 episodes were CRBSI, and 2 episodes were determined as bacterial colonization by retaining catheter methods. The 7 episodes were determined as bacterial colonization by overall estimation method, 6 episodes were correctly diagnosed, 1 episode was negative by retaining catheter methods, and 1 episode was determined as contamination. Quantitative blood culture had growth of bacteria only in chocolate medium. Conclusions The methods retaining the venous catheter can accurately detect the CRBSI.

Investigation on the effects of Adiponectin on Aldehyde oxidase-1 expression of hepatic cell lines in vitro

ZHANG Liang;CHEN Zhiqiang;WEN Jian

2012, 27(4): 275-279.

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Objective To investigate the influence of adiponectin（APN） on aldehyde oxidase-1 （AOX1） expression of HepG2 hepatoma cell lines and HL-7701 hepatic cell lines incubated by saturated fatty acid (SFA), and discuss the curative effect and possible mechanism of APN to protect non-alcohol fatty liver disease（NAFLD）. Methods Fatty HepG2 hepatoma cell lines and HL-7701 hepatic cell lines were incubated by 100 μmol/L SFA, and cells without being incubated by 100 μmol/L SFA were as controls. The fatty HepG2 hepatoma cell lines and HL-7701 hepatic cell lines were performed the treatment of APN (0.5,1.0,1.5 and 2.5 μg/mL), and cells without APN treatment were used as control group. Reverse transcription-polymerase chain reaction（RT-PCR） and Western-blot were utilized to detect the expressions of AOX1 gene and AOX1 protein. Results The AOX1 gene and AOX1 protein expressions of cells incubated by 100 μmol/L SFA were significantly higher than those of controls without being incubated by 100 μmol/L SFA (P<0.05), and the expression of HepG2 cells was significantly higher than that of HL-7701 cells (P<0.05). The AOX1 gene and AOX1 protein expressions of cells treated by APN were reduced significantly comparing to those without APN treatment (P<0.05)， which indicated that APN could decrease the expression effectively. However， the AOX1 expression of cells had no change, when the APN concentration reached 1.5 μg/mL. The AOX1 gene and AOX1 protein expressions of HepG2 cells decreased significantly higher than those of HL-7701 cells at the same APN concentration. Conclusions APN could reduce the expression of cells incubated by saturated fatty acid effectively， therefore it could protect hepatic cells from NAFLD.

The significance of heart-type fatty acid-binding protein for early diagnosis of acute myocardial infarction

LIN Gaogui;MENG Fanchao;ZENG Yunxiang;ZHAO Chun

2012, 27(4): 280-283.

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Objective To investigate the significance of heart-type fatty acid-binding protein (H-FABP） for the early diagnosis of acute myocardial infarction (AMI). Methods A total of 144 patients suffered chest pain as the main syndrome were enrolled in the study. The patients were AMI suspected (90 truth cases and 54 suspicion cases). By enzyme-linked immunosorbent assay (ELISA)， serum samples were detected respectively within 3 h and after 6 h of chest pain onset. The H-FABP, cardiac troponin I (cTnI) and creatine kinase isozyme-MB (CK-MB) were detected, and the sensitivity， specificity and diagnostic accuracy in the early diagnosis of AMI were compared. Moreover， 60 healthy subjects with similar age and sex were as the control group. Results The level of H-FABP［(56.13±19.17) ng/mL］ in AMI group was obviously higher than those in the suspected group ［(11.46±4.85) ng/mL］ and the control group ［(3.14±1.52) ng/mL］within 3 h (P<0.01). Furthermore， its seropositive rate (91.11%)，sensitivity (91.11%)，specificity (98.33%)， diagnostic accuracy (94.67%), positive predictive value (98.80%) and negative predictive value (88.24%) were obviously better than cTnI (13.33%,13.33%,96.67%,48.00%,85.71% and 43.48%) and CK-MB (21.11%,21.11%,95.00%,52.67%,86.36% and 48.80%)(P<0.05，P<0.01). The cTnI and CK-MB levels in AMI group were obviously higher than those in the suspected group and the control group after 6 h (P<0.01). Conclusions Within 3 h of AMI onset，H-FABP shows good sensitivity，diagnositic accuracy and negative predictive value. H-FABP can be used in the early diagnosis of AMI.

Associativity of Salusin-α and UⅡ Levels with Carotid Intima-Media Thickness in Patients with Essential Hypertension

JIANG Yonglin;JIANG Heping

2012, 27(4): 284-286.

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Objective To investigate the correlation between serum Salusin-α， urotensinⅡ(UⅡ) and carotid intima-media thickness(IMT) in patients with essential hypertension (EH). Methods A total of 100 patients with EH and 100 healthy controls in the same time were enrolled. Serum Salusin-α and UⅡ were detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay respectively. The IMT was detected by high-resolution ultrasound which was as an index of large vessel atherosclerosis (AS). The correlation of serum Salusin-α and UⅡ levels with IMT was analyzed. Results Serum UⅡ level and IMT were significantly higher in EH group than those in control group (P<0.01)，while Salusin-α level was significantly lower in EH group than those in control group(P<0.01). IMT was negatively correlated with serum Salusin-α(r=-0.527，P<0.01)， and positively correlated with serum UⅡ(r=0.612，P<0.05). Conclusions The change of serum Salusin-α and UⅡ has a correlation with carotid IMT， which suggests that Salusin-α and UⅡmay affect the resilience of arteries and participate the pathological process of AS. Serum Salusin-α and UⅡ may be new markers correlated with the severity of AS for clinical application.

Clinical significance of serum neutrophil gelatinase-associated lipocalin detection in early diagnosis of acute kidney injury

CAI Leiming;LI Qian;JIN Caifeng;TAN Longyi;LIAO Minlei;BAO Jie

2012, 27(4): 287-290.

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Objective To investigate the clinical significance of serum neutrophil gelatinase-associated lipocalin(NGAL) detection in early diagnosis of acute kidney injury(AKI). Methods A total of 60 inpatients in intensive care unit (ICU) and coronary care unit (CCU) without chronic kidney diseases were enrolled. Sera were collected before surgery and 6, 12 and 48 h after surgery respectively. There were 27 AKI patients and other 33 patients without AKI. The concentration of NGAL was determined by enzyme-linked immunosorbent assay (ELISA). Serum creatinine (SCr) was detected by Jaffé reaction method. Serum cystatin C (Cys C) was detected by Latex-enhanced immunoturbidimetry. AKI was defined as an increase in SCr of at least 50% from baseline within 48 h. The efficiency of NGAL in early diagnosis of AKI was evaluated by receiver operating characteristic (ROC) curve. Results Serum NGAL increased significantly 6, 12 and 48 h after surgery， while serum Cys C increased 12 and 48 h after surgery, and SCr increased 48 h after surgery in the AKI group. There was no obvious difference in the concentrations of NGAL, SCr and Cys C in the non-AKI group before and 6, 12 and 48 h after surgery (P>0.05). By ROC curve analysis, serum NGAL 6 h after surgery had an area under the curve of 0.51 ［95% confidence interval (CI): 0.25-0.78］， serum NGAL 12h after surgery had an area under the curve of 0.80 (95%CI: 0.58-0.95)， and serum NGAL 48 h after surgery had an area under the curve of 0.85 (95%CI: 0.61-0.99). 〖Conclusions Serum NGAL increases 6 h after surgery and predicts AKI accurately 12 h after surgery. For predicting AKI, serum NGAL is earlier than Cys C and SCr， and shows that serum NGAL can be as potential marker for the early diagnosis of AKI.

The establishment of double-antibody sandwich light initiated chemiluminescence assay for the determination of plasma myeloperoxidase

HAN Hairong;LIANG Ying;ZHAO Weiguo;SHEN Qian

2012, 27(4): 291-295.

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Objective To establish a quantitative double-antibody sandwich light initiated chemiluminescence assay (LiCA) for the determination of plasma myeloperoxidase (MPO) concentration in order to assist to diagnose cardiovascular diseases. Methods From 12 anti-MPO monoclonal antibody (McAb), the optimum double-antibody combination (one McAb coated on luminescence particles and the other McAb conjugated with biotin) was picked out by the checkerboard titration. The suitable reaction concentration and conditions were analyzed. Consequently, the repeatability, sensitivity, anti-interference and recovery rate of the assay were evaluated and compared with the classic CardioMPO^TMenzyme-linked immunosorbent assay (ELISA). Results Two McAbs were selected successfully. The quantitative MPO LiCA system was constructed by connecting the sensitizer particles with streptavidin. The results showed that the sensitivity was 1.76 ng/mL. The within-run and interday-run coefficients of variation (CV) of MPO LiCA were 2.73％ and 3.76％ respectively, and the mean recovery rate was 102.5%±8.6%. The correlation with CardioMPO^TMELISA was high (r=0.961，P<0.01). The normal reference range was 13.16-128.79 ng/mL. Conclusions A quantitative LiCA for the determination of MPO is successfully established. MPO LiCA with good sensitivity, stability, long reagent validity, simple and convenience shows a better application prospect in assisting to diagnose cardiovascular diseases.

The changes and relationship with sex hormone, cytokine and bone metabolic marker of postmenopausal osteoporosis patients

YI Weilian;LIAO Dequan;LIN Baiyun;XIAO Yajing;YUAN Hanyao

2012, 27(4): 296-298.

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Objective To investigate the correlation of sex hormone, cytokine and bone metabolic index in postmenopausal osteoporosis (POP) patients. Methods According to the results of the lumbar spine bone mineral density (BMD), the subjects were classified into 3 groups: POP group (48 cases), non-osteoporotic postmenopausal (NOP) group (32 cases) and premenopausal healthy group (30 cases). By radioimmunoassay, chemiluminescence and enzyme-linked immunosorbent assay (ELISA), sera of all 110 cases were tested for the levels of estradiol (E2), interleukin-6 (IL-6), insulin-like growth factor-1 ( IGF-1), bone alkaline phosphatase (BAP) and osteocalcin middle amino terminal (N-BGP), and the correlation was analyzed. Results The levels of IL-6, N-BGP and BAP of POP group were increased and higher than those of NOP group and control group (P<0.01), while the BMD and the levels of E2 and IGF-1 of POP group were lower than those of NOP group and control group (P<0.01). IGF-1 with BMD and E2 showed a significant positive correlation (r=0.572 and 0.436, P<0.01), and IGF-1 was negatively correlated significantly with age (r=-0.541, P<0.01). IL-6 with BMD and E2 was negatively correlated (r=-0.584 and -0.435, P<0.05), and IL-6 with N-BGP and BAP was positively correlated (r=0.546 and 0.354, P<0.05). BMD with IL-6, age, N-BGP and BAP was negatively correlated (P<0.05), and BMD with E2 and IGF-1 was positively correlated (P<0.05). Conclusions The levels of IL-6, N-BGP, BAP, E2 and IGF-1 may be indices of clinical diagnosis and treatment of POP.

Clinical Application of WBC Differential of CellaVision DM96 System

HUANG Jibin;ZENG Tingting;GUO Manying;HAN Xiuhua;JIANG Hong

2012, 27(4): 299-303.

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Objective To verify and evaluate the performance of CellaVision DM96 system (DM96). Methods A total of 422 peripheral blood samples were collected and prepared by automated smear and stain system， and then analyzed by DM96 white blood cell differentiation. The results of white blood cell differentiation by DM96 were compared with those by manual microscopy. The correlation of the results by DM96 and manual microscopy was analyzed by linear regression analysis. The coincidence rate of identifying cells based on original and enlarged reference cell libraries was calculated. The ability to identify important abnormalities through analyzing 110， 210，310 and 410 cells and the time cost were calculated. Results The correlation coefficients of neutrophils and lymphocytes were 0.91 and 0.88 between DM96 and manual microscopy respectively， while the correlation coefficient of monocytes was only 0.31. DM96 agreed 27.56%，37.96%，41.85%，45.12% and 29.76% for pre-differentiation of promyelocytes， myelocytes， metamyelocytes ， blast cells and variation lymphocytes based on original reference cell library，and 33.23%，56.86%，48.33%，58.08% and 31.25% based on enlarged reference cell library. The ratios of identifying important abnormalities were 87.50%， 92.65%, 94.85% and 97.59% when analyzing 110， 210， 310 and 410 cells. It took (2.50±0.41) min per case by manual microscopy， (1.98±0.20) min per case by DM96 and (0.91±0.14) min per case by manual confirming. Time costs of analyzing based on original and enlarged reference cell libraries were (2.01±0.39) and (2.09±0.54) min per case. With the numbers of analyzed cells increasing， time costs increased simultaneously. Conclusions The clinical application of DM96 could improve the automatization and standardization of white blood cell differentiation.

The polymorphism determination of T1128C in neuropeptide Y gene among Han population in Shanghai

MIAO Yingxin;GAN Jiemin;SHI Hong;CHEN Jie;ZHOU Xiaoqian;ZHAO Hu

2012, 27(4): 304-307.

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Objective To investigate the distribution frequency of T1128C (Leu7Pro) single-nucleotide polymorphism (SNP) in neuropeptide Y (NPY) gene among Han population in Shanghai. Methods Polymerase chain reaction (PCR)-ligase detection reaction (LDR) was performed to detect the T1128C polymorphism in NPY gene among 867 Han people in Shanghai. A pair of primers and 3 probes were designed according to target DNA sequence. The LDR was performed after PCR amplification, and the genotypes were analyzed according to the length of fragments of LDR products on a sequencer by electrophoresis. Results All 867 subjects showed T1128T(Leu7Leu) but no T1128C(Leu7Pro) or C1128C(Pro7Pro) in the NPY gene.〖WTHZ〗 Conclusions〖WTBZ〗 There is difference of T1128C polymorphism in NPY gene among populations with different genetic and geographic backgrounds. There is no T1125C (Leu7Pro) polymorphism in NPY gene among Shanghai Han population. There is no correlation of T1128C polymorphism in NPY gene with coronary heart disease among Shanghai Han population.

Study on the apolipoprotein E gene polymorphism in Li patients with cardiovascular and cerebrovascular disease

WANG Yongqing;WU Chanjuan;YAO Min;ZHANG Ying′ai;ZHENG Lingling

2012, 27(4): 308-310.

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Objective　To study the apolipoprotein E (apo E) gene polymorphism in Li population patients with cardiovascular and cerebrovascular disease, and investigate the relationship of polymorphism with cardiovascular and cerebrovascular disease. Methods　The ligase detection reaction (LDR) technique was used to detect apo E gene polymorphism in 150 patients with cardiovascular and cerebrovascular disease (50 patients with hypertension， 50 patients with cerebral infarction and 50 patients with coronary artery disease) and 50 healthy controls. The results were analyzed. Results　In the disease group and control group, ε3/3 genotype frequency was the highest. The ε2 allel frequency in cerebral infarction group was lower than those in hypertension group, coronary artery disease group and control group (P<0.05). There was no statistical significance in the allels among the hypertension, coronary artery disease and control groups (P>0.05). Conclusions　The apo E ε2 allel may be a major factor to reduce the incidence of cerebral infarction in Li population.

Two quality standard in the application of clinical chemical analysis performance evaluation.

QIU Haiwen;ZHANG Yuanyuan;CHEN Min

2012, 27(4): 311-315.

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Objective To investigate the application of 2 quality standards in clinical chemical analysis performance evaluation. Methods The internal quality control imprecision and external quality control average deviation of our laboratory (HITACHI analysis system) were selected. The imprecision and inaccuracy of complete test system (Modular PPI analysis system) approved by the Food and Drug Administration (FDA) were selected. The allowable total error (TEa) according to the Clinical Laboratory Improvement Amendment (CLIA′88) were made by the biological variation (BV). Making use of Westgard performance evaluation decision graph, each detection system analysis performance meeting the prescribed quality requirements or not was analyzed. The analysis performance detection rate and applicability of Modular PPI analysis system with 2 quality standards were analyzed and evaluated. Results In 23 items，with CLIA′88 allow quality standards and requirements, the HITACHI system′s urea (Ur) and chlorine (Cl^-) determination methods can not satisfy the quality requirements， total protein (TP)， creatine kinase (CK) and uric acid (UA) belonged to the critical levels，total bilirubin (TBil)，alkaline phosphatase(ALP), total cholesterol(TC)，natrium (Na⁺) and magnesium (Mg) analysis performances were good，and the other 11 item analysis system performances were excellent. In the Modular PPI system, project analysis performance can be accepted. Meanwhile, the excellent rate，good rate，edge rate and discrepancy rate were 75%， 15%，10% and 0%，and for the BV "low limit"，"appropriate" and "ideal" quality standards, its discrepancy rates were 21%, 48% and 65%. Conclusions Westgard performance evaluation decision graph for evaluating the performance of biochemical detection system is accurate， simple and suitable for clinical laboratory application， and CLIA′88 quality requirement is more suitable than the current laboratory quality requirement. BV quality requirements are suitable for a part of excellent performance items.

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