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Troponin and acute coronary syndromes:more clinical studies needed

Chinese Journal of Laboratory Medicine.2001;0(05):-.

Monitoring cardiac troponin Ⅰ(cTnI)or T(cTnT)for the detection of myocardial injury and for risk stratification in acute coronary syndromes(ACS)has been endorsed by the laboratory medicine community(IFCC,AACC,NACB)and the cardiology community(ESC and ACC).The clinical laboratories has long been argued on the best cut-point values for troponin assay-some believe it should be set at the 99~(th)percentile of the reference population,while others believe it should be at the level where the particular assay's coefficient of variation(CV)is less than 10%,with the 10% CV varying by test manufacturer.Besides,there are numerous cTnI measurement systems,confusion regarding cTnI measurements has resulted large variability demonstrated among the cTnI assays.The aim of this opinion is to reflect on some concepts related to using cTn in clinical practice.

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Detecting extended-spectrum beta-lactamases in Escherichia coli and Klebsiella pneumoniae.

Qinglian KONG ; Xiujuan YU ; Ping YAO

Chinese Journal of Laboratory Medicine.2000;0(06):-.

Objective To detect ESBLs in E.coli and K.pneumoniae with appropriate method.Methods The suspiciously produced ESBLs were detected by standard screen test among 110 E.coli and 84 K.pneumoniae. ESBLs possessing strains were confirmed by E test, single disc diffusion test with clavulanic acid or sulbatan, and two kinds of double disc synergy test. Results There was a significant difference between the detected rate of ESBLs in E.coli (16/26) and K. pneumoniae (4/20) by E test( P 0.05). The single disc diffusion test with clavulanic acid or sulbatan was applicable to confirm ESBLs in K. pneumoniae (12/20). Conclusions Single disc diffusion test is a sensitive, convenient and inexpensive method to confirm ESBLs in E.coli and K.pneumoniae in clinical laboratory.

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Detection of extended-spectrum-?-lactamases with VITEK-AMS and double-disk synergy test

Tieli ZHOU ; Meijie DAI ; Shengying CHI

Chinese Journal of Laboratory Medicine.2000;0(06):-.

Objectives To investigate the reliability of VITEK AMS for the detection of strains of E.coil and K.pneumoniae that produce ESBLs, and to determine the prevalence in our hospital of the two strains producing ESBLs.Method VITEK GNS 506 was used to detect ESBLs, and was compared with double disk synergy test. Results Among 48 strains of K.pneumoniase detected using VITEK GNS 506, 24 strains were found to be ESBLs positive, and 24 strains ESBLs negative. The result was consistent with that of double disk synergy test. Among 102 strains of E.coli detected using VITEK GNS 506, 41 strains were found to be ESBLs positive. The result was consistent with that of double disk synergy tests. 61 strains were found to be ESBLs negative detected using VITEK GNS 506. Among them, 19 strains were found to be ESBLs positive.Conclusion Detection ESBLs using VITEK AMS has some limitations in our area. Simple and reliable method should be used more with indicates for detection of ESBLs suitable for routine screening.

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Detection of ureaplasma urealyticum by polymerase chain reaction microplate hybridization and anti Uu susceptibility test

Xiaotao ZHAO ; Zheng ZHANG

Chinese Journal of Laboratory Medicine.2000;0(06):-.

Objectives To establish a sensitive and special method for the detection of Ureaplasma urealyticum(Uu) using PCR microplate hybridization (PCR MPH). Methods A primer of ureasea gene was labeled by biotin. The amplification product was captured on streptavidin coated microplates, then products were quantified by hybridization with a digoxigenin labeled internal oligonucleotide probe. After revelation with an anti digoxigenin alkaline phosphatase coupled antibody(anti DIG AP), the amount was determined by optical reading. At the same time, PCR MPH was compared with Bio Merieux Mycosplasma IST. Results A method of PCR MPH for detecting Uu DNA was established. The morbidity among three groups for detecting 158 clinical samples was analysed. 65 were detected by PCR MPH and 56 by culture.Conclusion The results showed that this assay is rapid, sensitive, specific, and accurate, and is of value in clinical therapy.

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The von Clauss method for fibrinogen assay and its influential factors

Feng CHENG ; Dan WANG ; Xiaohui ZHU

Chinese Journal of Laboratory Medicine.2000;0(06):-.

Objectives To study the influential factors of the von Clauss method and to promote the method for fibrinogen assay in clinical laboratories in China. MethodsKits of foreign and domestic products were used. Calibration curves for fibrinogen determination were established, and calibration plasma was used for comparison. Effects of pH value, thrombin activity, and temperature on the measurement were studied.Results The reproducibility of the von Clauss method using domestic thrombin and related reagents was excellent, the within day CV being 0.039 1, and the between day CV 0.049 7. The percentage variations of calibration plasma determination, with the foreign fibrinogen assay kits using domestic reagents, were +3.17% , +4.76%, and -6.03%, respectively. Determination of a lyophilized plasma using the two sources reagents showed similar results.Conclusions Thrombin and relevant reagents produced in China could be used as a substitute for kits purchased abroad, which would promote the use of von Clauss method in clinical laboratories of China.

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Design of method comparison study and bias estimation for albumin assays

Jian GUO ; Jiehong XIE ; Haijian ZHAO

Chinese Journal of Laboratory Medicine.2000;0(06):-.

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

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Influence of nucleic acid extraction from serum with two methods on HBV DNA detection by fluorescence quantitative PCR (TaqMan)

Jinming LI ; Lunan WANG ; Wei DENG

Chinese Journal of Laboratory Medicine.2000;0(06):-.

Objective To investigate the influence of nucleic acid extraction by boiling the serum samples and purifing nucleic acid from serum on HBV DNA detection by PCR. Methods The DNA templates from 2 serum samples with different HBV DNA concentration and 3 serum samples with hemolysis and non hemolysis for PCR were prepared by two kinds of method, i.e. by boiling the serum to release HBV DNA from the virus particles and by purifing the nucleic acid from serum. Then,the difference of reproducibility and HBV DNA quantitative values by PCR using the templates prepared by the two nucleic acid extraction methods was compared.Results HBV DNA quantitative values by PCR using the templates purified from the two samples were more reproducible( CV : 0.2165 and 0.3262, respectively) than those using the templates prepared by boiling the serum ( CV : 0.0699 and 0.1092, respectively). Moreover, when the concentration of hemoglobin in serum samples was higher than 5.89 ?g/L, the HBV DNA quantitative values by PCR using the templates purified from the samples were about one hundred fold more than those using the templates prepared by boiling the serum ( P

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Anti-TK1 antibody in detecting thymidine kinase 1 in serum of breast tumor patients

Yan LI ; Pingan ZHANG ; Li ZOU

Chinese Journal of Laboratory Medicine.2000;0(06):-.

Objective To evaluate the value of immunoblotting in the detection of serum thymidine kinase 1 (TK1) in breast tumor patients. Methods TK1 in sera from breast tumor patients, non tumor patients and healthy volunteers were determined with immunoblotting assay.Results The levels of TK1 in breast cancer patients before operation, patients with benign breast tumor, non tumor patients, and healthy volunteers were (39.3? 33.9), (3.7?3.9), (1.7?0.5) and (1.4?0.5) pmol/L respectively. The value of TK1 was higher in malignant and benign breast neoplasms patients than in healthy volunteers. However, no significant difference in the TK1 level was found between breast cancer patients without metastasis after operation, non tumor patients, and healthy volunteers.Conclusions The immunoblotting of TK1 by anti TK1 antibody is a sensitive and specific assay. It can be used to monitor and predict the efficiency of the therapy as well as the breast neoplasms progression during clinical course.

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sCD44v6 in the diagnosis of colorectum cancer

Qishui OU ; Bin LAN ; Haibing XU

Chinese Journal of Laboratory Medicine.2000;0(06):-.

Objective To explore the role of sCD44v6 in the diagnosis of colorectum cancer. Methods ELISA was used to detect the concentration of sCD44v6 of 81 patients after or before surgery.Results The level of sCD44v6 in the group of colorectum cancer was 200.5?31.1 ng/ml, which was higher than that of normal controls (45.6?9.5ng/ml) and that of patients with coloritis (48.3?7.6ng/ml). The concentration of sCD44v6 in the group of colorectum cancer with metastasis(280.1?26.9ng/ml) was higher than that of the group without metastasis(160.9?34.4ng/ml). The level of sCD44v6 was related to tumor burden.Conclusions The level of sCD44v6 is closely related to colorectal cancer and its metastasis. It would be useful in the diagnosis, prognosis, and forcasting of metastasis.

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cagA +cagE + Helicobacter pylori induced interleukin 8 secretion in gastric epithelial cells and its dependence on protein tyrosine kinase activation

Jieli WU ; Xingxiang HE ; Guobin WANG

Chinese Journal of Laboratory Medicine.2001;0(01):-.

Objective To study the effect of different cag pathogenicity island in Helicobacter pylori ( Hp ) from Chinese patients and various kinase inhibitors on Hp induced interleukin 8 (IL 8) secretion in Chinese gastric epithelial cells. Method Chinese gastric epithelial cells(MGC 803) were cocultured with Chinese clinical cagA +cagE +, cagA +cagE -,cagA -cagE +,cagA -cagE - Hp in vitro, respectively. At the end of culture, IL 8 protein secretion was assayed by ELISA. The effect of the inhibitor of protein kinase A(PKA), C, G, protein tyrosine kinase(PTK) was analyzed on IL 8 protein secretion in gastric epithelial cells by Hp stimulation. Results cagA +cagE + Hp induced IL 8 protein secretion higher than cagA + cagE - or cagA -cagE + Hp , but cagA -cagE - Hp didn′t increase IL 8 protein secretion in gastric epithelial cells. Further studies with gastric epithelial cell showed that IL 8 protein secretion induced by cagA +cagE + Hp was blocked by the PTK inhibitor herbimycin A but not by PKA inhibitor H7, PKC inhibitor calphostin C, and PKG inhibitor KT5823. Conclusion cagA +cagE + Hp significantly increases IL 8 protein secretion and it depends on PTK activation in gastric epithelial cells.

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