Table of Content

28 February 2015, Volume 30 Issue 2

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Orignal Article

The prospects of mass spectrometry application on microbiology

LUO Yanping

2015, 30(2):  97-100.  DOI: 10.3969/j.issn.1673-8640.2015.02.001

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The matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) technique can be used to generate protein fingerprint signatures from whole bacterial cells. By comparing these fingerprints to a database of reference spectra by the various algorithms, bacteria can be rapidly identified. MALDI-TOF MS can be used for accurate and rapid identification of various microorganisms, such as common bacteria, Nocardia, Mycobacterium, anaerobe, yeast and filamentous fungi. MALDI-TOF MS have recently been widely introduced. The characteristics of one review and four papers published in the special subject about MALDI-TOF MS were reviewed. The development of studies in MALDI-TOF MS will play an important role in the application of mass spectrometry in clinical microbiological laboratories.

Application of MALDI-TOF MS technology in clinical microbial determination

DAI Yingxin, LI Min

2015, 30(2):  102-107.  DOI: 10.3969/j.issn.1673-8640.2015.02.002

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Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) is one kind of soft ionization mass spectrometry, which is suitable for the analysis of bio-macromolecule, such as protein. It can be used for microbial identification by mapping the microbial protein mass spectra. Since MALDI-TOF MS technology is well-known for its rapidity, accuracy, high sensitivity and high throughput, it has been applied in bacterium and fungus identifications. MALDI-TOF MS also has a good performance in the field of scientific research, including resistance analysis, bacterium typing and virulence study,and there is still a large space for development. In this paper, we reviewed the application of MALDI-TOF MS technology in microbial identification and scientific research.

Application of MALDI-TOF MS for identifying bacteria directly in midstream urine samples

WANG Lin, LI Jiaping, CHEN Gongxiang, ZHANG Rong

2015, 30(2):  108-112.  DOI: 10.3969/j.issn.1673-8640.2015.02.003

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To construct rapid identification method for clinical laboratory with the use of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) for the direct identification of bacteria in midstream urine samples.

A total of 692 midstream urine samples were cultivated on agar,and were identified by MALDI-TOF MS and Vitek 2 Compact automated microbial amalyslis system (Vitek 2 Compact). Direct identification of midstream urine samples were also conducted by MALDI-TOF MS after preparation.

Of the 692 midstream urine samples, 98 samples were positive for one kind of bacteria (>10⁴ CFU/mL). The overall detection rate of direct identification by MALDI-TOF MS was 75.5%,and 74 isolates were identified,among which 39 isolates were Escherichia coli with the detection rate of 79.6%,23 isolates were Enterococcus faecalis with the detection rate of 100.0%,and 5 isolates were Morganella morganii with the detection rate of 100.0%. Among the 64 midstream urine samples with multiple microorganisms,44 samples were detected with a dominant bacteria by direct identification. The consistency rate was 100.0% among the results by direct identification,the identification of colony by MALDI-TOF MS and Vitek 2 Compact.

MALDI-TOF MS could be applied to identify directly bacteria in midstream urine samples with high detection rate and accuracy, and MALDI-TOF MS also shows certain performance in identifying samples containing multiple bacteria. The identification results could be 42 h earlier available than regular methods. It is a revolutionary improvement for the clinical identification of bacteria in urine samples.

Direct identification of microorganisms from blood culture by MALDI-TOF MS combined with separation gel tube

CHEN Feng, LI Yuanrui, HUANGFU Yuchan, TAO Xiaoqin, LIU Ying

2015, 30(2):  113-121.  DOI: 10.3969/j.issn.1673-8640.2015.02.004

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To evaluate the direct identification of microorganisms from blood culture by a separation gel tube-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and its identification rate.

A total of 491 cases with positive blood culture were collected. The bacteria were enriched and purified directly from blood culture bottle by separation gel tube, and then the bacteria were identified by MALDI-TOF MS. Traditional culture and identification processes were performed simultaneously, and the cultured pure colonies were identified by Vitek 2 Compact automated microbial analysis system (Vitek 2 Compact). The identification of bacterial isolates by MALDI-TOF MS was compared with that of traditional biochemical testing, and discrepancies were resolved by gene sequencing.

A total of 491 positive cultures were determined, representing 30 genera and 64 species or groups. The method demonstrated 73.6% and 3.7% concordance to species and genus identification rates in monomicrobial culture (462 cases). The correct species and genus identification rates were 84.0%(147 cases) and 2.9%(5 cases) in Gram-negative bacteria(175 cases), were 75.3%(189 cases) and 4.4%(11 cases) in Gram-positive bacteria(251 cases), and were 19.4%(7 cases) and 2.8% (1 case) in Candida species(36 cases). In 29 cases of complex bacterial infection, the correct species and genus identifications rates in one of the multiple bacteria were 79.3% (23 cases) and 10.3% (3 cases). Species identification rates of bloodstream infection with common pathogens were 83.3%-96.9%.

This separation gel tube-based MALDI-TOF MS for the direct identification of blood culture pathogens is established. The identification rate of common pathogens in bloodstream infection by MALDI-TOF MS is higher compared with that by traditional culture identification processes. In addition, it is rapid, easy and cost-effective, and it is a recommendable method for clinical microbiology laboratories.

Comparison on the identification abilities of Microflex^TM MALDI-TOF MS and Vitek 2 Compact automatic microbial analysis system for Enterobacteriaceae

LIU Ying, YU Jing, CHENG Feng, LIU Jingxian, LI Yuanrui, HUANGFU Yuchan, SHEN Lisong

2015, 30(2):  122-127.  DOI: 10.3969/j.issn.1673-8640.2015.02.005

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To compare the identification abilities of Microflex^TM matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and Vitek 2 Compact automatic microbial analysis system (Vitek 2 Compact) for Enterobacteriaceae.

A total of 545 isolates of Enterobacteriaceae from a variety of clinical and quality control isolates were identified by Microflex^TM MALDI-TOF MS and Vitek 2 Compact. The isolates were confirmed by 16S rDNA gene sequencing, and Salmonella was identified by serum agglutination test, if the identification results of the 2 systems didn't match.

The coincidence rates of 545 isolates of Enterobacteriaceae identified by Microflex^TM MALDI-TOF MS were 97.1% to the species level and 2.9% to the genus level,respectively. The coincidence rates of 545 isolates of Enterobacteriaceae identified by Vitek 2 Compact were 83.3%,13.9% and 2.2% to the species, group and genus levels, respectively. The false rate was 0.2%, and the un-identification rate was 0.4%.

The identification coincidence rate of Enterobacteriaceae by Microflex^TM MALDI-TOF MS is higher than that of Vitek 2 Compact. Using Microflex^TM MALDI-TOF MS to identify Enterobacteriaceae is fast,simple to operate and low-cost,and it can be used in routine rapid identification of Enterobacteriaceae in clinical microbiology laboratories.

Rapid identification of Yeast like fungi outside of Candida albicans isolated form blood culture by two MALDI-TOF MS systems

HUANG Shenglei, HU Bijie, CHEN Rong, ZHOU Chunmei

2015, 30(2):  128-131.  DOI: 10.3969/j.issn.1673-8640.2015.02.006

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To evaluate the application of 2 matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), Autoflex MALDI-TOF MS and Vitek MALDI-TOF MS, for the identification of Yeast like fungi outside of Candida albicans isolated from blood culture.

A total of 59 isolates were collected from blood culture from October 2010 to February 2013 among inpatients in Zhongshan Hospital, Fudan University. The results of fungus internal transcribed spacer (ITS) sequencing were as golden standards. The correct identification rates of Autoflex MALDI-TOF MS, Vitek MALDI-TOF MS and API 20C AUX were analyzed comparatively.

The 59 isolates included 11 types of fungi, namely, Candida parapsilosis complex(30 isolates, 50.8%), Candida tropicalis(11 isolates, 18.6%), Candida glabrata(8 isolates, 13.6%), Candida krusei( 2 isolates, 3.4%), Wickerhamomy cesanomalus(2 isolates, 3.4%), Cryptococcus neoformans(2 isolates, 3.4%), Candida iusitaniae(1 isolate, 1.7%), Candida guilliermondii (1 isolate, 1.7%), Trichosporon asahii(1 isolate, 1.7%) and Saccharomyces cerevisiae (1 isolate, 1.7%). The correct identification rates by Autoflex MALDI-TOF MS, Vitek MALDI-TOF MS and API 20C AUX were 93.2%, 83.1% and 79.7%, respectively, and there was no statistical significance between the 2 MALDI-TOF MS systems (P=0.155). The correct identification rates for yeast like fungi outside of Candida albicans by Autoflex MALDI-TOF MS, Vitek MALDI-TOF MS and API 20C AUX were 100.0%, 84.9% and 84.9%, respectively, and there was statistical significance between the 2 MALDI-TOF MS systems(P=0.006). The correct identification rates of rarely clinical encountered fungi by Autoflex MALDI-TOF MS, Vitek MALDI-TOF MS and API 20C AUX were 33.3%, 66.6% and 33.3%, respectively.

MALDI-TOF MS system is rapid, simple and accurate compared with current routine biochemical identification methods. The performance of Autoflex MALDI-TOF MS is similar with that of Vitek MALDI-TOF MS for the identification of isolated fungi from blood culture. The performance of Autoflex MALDI-TOF MS is better than that of Vitek MALDI-TOF MS for the identification of yeast like fungi outside of Candida albicans.

Immunophenotyping and bone marrow hematology analysis in 55 cases of non-Hodgkin's lymphoma

YANG Limiao, ZHANG Yaping, YANG Hongle, XING Jiangtao, HU Rui, ZHU Yun

2015, 30(2):  132-136.  DOI: 10.3969/j.issn.1673-8640.2015.02.007

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To understand the progression of non-Hodgkin's lymphoma through bone marrow morphology, and to find out the application significance of immunohistochemistry in the diagnosis of non Hodgkin's lymphoma in order to determine the clinical stage.

The monoclonal antibody to some specimens of non-Hodgkin's lymphoma was used for immunohistochemical detection, and bone marrow morphology was performed in 55 lymphoma patients.

By immunohistochemistry, the positive rate of T cell lymphoma associated antibody CD3 was 85.00%, and that of CD45RO was 90.47%, that of B cell lymphoma associated antibody CD20 was 89.65%, and that of CD79a was 88.90%. By bone marrow morphology, 9 cases (16.36%) had bone marrow involvement, 7 cases (12.72%) progressed to lymphoma-leukemia period. There were 19 patients whose lymphoma cells <5%, for those patients, a dynamic observation should be on, which can closely observe the morphological changes in bone marrow.

Immunohistochemistry can determine the type of T or B lymphoma, and it is a supplement to the histomorphological diagnosis of lymphoma, so that a more accurate and reliable diagnosis can be made. Bone marrow morphology can determine whether the patients' bone marrow is involved and whether it is already advanced in leukemia period, therefore the clinical stage of lymphoma can be identified.

Difference cause analysis on thrombocytopenia by different bacterial infections in patients with sepsis

YAN Feng, REN Zhenhuan, ZHOU Ying

2015, 30(2):  137-140.  DOI: 10.3969/j.issn.1673-8640.2015.02.008

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To analyze primarily the difference causes of thrombocytopenia by different bacterial infections in patients with sepsis, to distinguish the biochemical characteristics of Gram-positive bacteria (G⁺) and Gram-negative bacteria (G^-), and to provide the reference for the clinical early prevention of multiple organ failure and reduce the mortality rate.

A total of 155 sepsis patients, 56 patients with non-sepsis (infection group) and 43 non-infected controls were enrolled, and their platelet (PLT), procalcitonin (PCT), C reactive protein (CRP), white blood cell (WBC), the percentage of neutrophils (NE) and Bcl-xL protein levels were determined. The results were analyzed statistically. Blood culture was performed simultaneously, 75 cases of blood culture-positive sepsis patients were classified into G⁺ and G^- groups. Their PLT, PCT, Bcl-xL protein and different PCT cross-section percentages were analyzed.

Biochemical parameters of sepsis group had obvious difference with those of control group (P<0.01). Compared with infection group, serum PLT, PCT, CRP and Bcl-xL protein levels were significantly different (P<0.01), and WBC and the percentage of NE had no statistical significance (P> 0.05). In infection group, PCT, CRP, WBC, NE percentage had statistical significance compared with those in control group (P<0.01), but the differences of PLT and Bcl-xL protein were not significant (P> 0.05). For G⁺ and G^- groups in sepsis patients, when PCT <2.0 ng/mL, there was no statistical significance (P>0.05). When 2.0 ng/mL≤PCT≤10.0 ng/mL, the percentage in G⁺ group was significantly higher than that in G^- group (P<0.01). When PCT ≥ 10.0 ng/mL, the percentage in G^- group was significantly higher than that in G⁺ group (P<0.01). PLT and PCT levels in G^- group were lower than those in G⁺ group (P<0.01).

The reduction of PLT and the increase of PCT are related to the apoptosis of PLT caused by Bcl-xL protein. G^- sepsis is easier to start PLT apoptosis than G⁺ sepsis with higher risk.

The variation and its clinical significance of Th1/Th2 and CD28⁺ cells from peripheral blood in patients with chronic glomerulonephritis

ZHOU Yunjiao, LI Peng, ZHANG Lihong, QIAN Yingjun, NIU Jianying

2015, 30(2):  141-144.  DOI: 10.3969/j.issn.1673-8640.2015.02.009

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To investigate the possible mechanism of cellular immunology in the process of chronic glomerulonephritis (CGN) by the detection of Th1/Th2 ratio and CD28⁺ cell percentages from peripheral blood in patients with CGN.

Th1/Th2 ratio and CD28⁺ cell percentages were detected by flow cytometry from 50 patients with CGN and 30 healthy subjects (control group). SPSS 16.0 software was used to analyze the results.

Compared with control group, the percentages of CD3⁺CD4⁺ cells and CD4/CD8 ratio of patients with CGN decreased significantly (P<0.05). Although there was no statistical significance in the percentages of Th1 and Th2 cells (P>0.05), yet Th1/Th2 ratio increased significantly (P<0.05). The percentages of CD28⁺ and CD4⁺CD28⁺ cells decreased significantly (P<0.01).

T cell immune function is disorder, and activation is restricted in patients with CGN. Their cellular immune function is low, and there exists imbalance between Th1 and Th2 cells. The detection of Th1/Th2 ratio and percentages of CD28⁺ cells from peripheral blood in patients with CGN is valuable to understand the immune state so as to provide the theoretical basis for immune therapy.

The band patterns of Western blotting in different clinical stages of HIV-1 infection/AIDS patients

GUO Chuan, JI Linying, WU Yaobo, LIN Jiemin, CHEN Wan

2015, 30(2):  145-148.  DOI: 10.3969/j.issn.1673-8640.2015.02.010

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To analyze the change of anti-virus antibodies in serum of patients with different clinical stages of human immunodeficiency virus 1 (HIV-1) infection/ acquired immune deficiency syndrome (AIDS), and to investigate the band pattern characteristics of Western blotting (WB).

CD4⁺ T cell counting was determined in the part of HIV-1 confirmed cases in Shantou from 2009 to 2012. According to the results, these cases were classified into 3 groups, primary-stage infection group (CD4⁺ ≥500 /mm³), middle-stage infection group (CD4⁺ ≥ 200/mm³ and CD4⁺ <500 /mm³) and AIDS group (CD4⁺ <200 /mm³). The WB results of the 3 groups were analyzed and performed to determine the positive rates of antibodies against viral proteins (p17, p24, p31, p39, gp41, p51, p55, p66, gp120 and gp160) and the band patterns of WB.

The antibodies against viral protein p24, p31, gp41, p51, p66, gp120 and gp160 showed high total positive rates (98.2%, 92.0%, 90.8%, 89.5%, 93.3%, 98.7% and 97.5%, respectively) , while the positive rates had no obvious difference among the 3 stages (P>0.05). The antibodies against viral proteins p17, p39 and p55 showed low positive rates (68.9%, 49.1% and 42.5%, respectively). The positive rate of protein p17 showed an obvious declining between primary-stage infection groups (91.5%) and middle-stage infection group 62.8% (P<0.01)as well as AIDS group 59.3% (P<0.01). All bands, missing p39+p55 and missing p17+p39+p55 were the common band patterns of WB among the 3 groups. Primary-stage infection group showed the highest appearance rate (60.8%) of all bands but the lowest appearance rate (6.5%) of missing p17+p39+p55.

Turning negative of antibody against viral protein p17 could be used as a potential distinguish parameter to forecast the disease progression of HIV-1 from primary-stage infection to middle-stage infection group/AIDS group.

Analysis of bone metabolism balance in chronic renal failure patients with 25-hydroxy vitamin D deficiency

WANG Ying, SHEN Jianxiong, XU Min, XUE Zhizhong

2015, 30(2):  149-151.  DOI: 10.3969/j.issn.1673-8640.2015.02.011

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To investigate the characteristics and relationship of bone metabolism in chronic renal failure patients with 25-hydroxy vitamin D deficiency.

A total of 80 chronic renal failure patients with 25-hydroxy vitamin D≤37.5 nmol/L were enrolled as study group, and 52 healthy subjects with 25- hydroxy vitamin D>75.0 nmol/L were as enrolled as control group. Their peripheral blood osteocalcin, total procollagen type Ⅰ amino terminal peptide (tP1NP), C-end peptide beta isomerization (β-CTx), parathyroid hormone (PTH), bone alkaline phosphatase (BAP), calcium, magnesium, phosphorus and ferrum were determined. The results were analyzed statistically.

Compared to control group, tP1NP, osteocalcin, β -CTx, PTH and phosphorus in study group were higher than those in control group (P<0.01). The other parameters had no statistical significance (P>0.05). 25-hydroxy vitamin D levels and calcium were positively correlated [correlation coefficient (r)=0.33, P<0.05].

The characteristics of bone metabolism in chronic renal failure patients with 25-hydroxy vitamin D deficiency are enhancing the activity of osteoclasts and bone resorption significantly. It is more important to detect bone turnover rate than calcium in patients with chronic renal failure.

Comparison on the detection significance of blood glucose and glycated serum protein among traumatic fracture patients

LI Xiaonan, LIU Junquan

2015, 30(2):  152-155.  DOI: 10.3969/j.issn.1673-8640.2015.02.012

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To study the fact of blood glucose (Glu) rising being the result of diabetes mellitus or post-traumatic stress response of body through detecting the changes of Glu and glycated serum protein (GSP).

The 87 patients with acute traumatic fracture were classified into 3 groups according to the presence or absence of type 2 diabetes mellitus, traumatic fracture with diabetes group(17 cases), traumatic fracture with unknown diabetes group (22 cases diagnosed as diabetes after hospitalization) and traumatic fracture without diabetes group (48 cases). The Glu and GSP levels of each group were measured by glucose oxidase method and fructosamine assay when hospitalized and cured. A total of 30 healthy subjects were enrolled as healthy controls.

The Glu levels of acute traumatic fracture groups were significantly higher than that of control group (P<0.05), especially traumatic fracture with unknown diabetes group, and there was statistical significance (P<0.05). The level of GSP of traumatic fracture with unknown diabetes group was higher than those of the other 3 groups (P<0.05), while the GSP levels of the other 3 groups had no statistical significance (P>0.05). The Glu level of acute traumatic fracture groups returned to normal level when cured, while the GSP level of traumatic fracture with unknown diabetes group increased and was significantly higher than those of the other 3 groups (P<0.05), due to high level of Glu. The Glu level of traumatic fracture without diabetes group when cured was significantly lower than that when hospitalized (P<0.05), while there was no significant difference for GSP level (P>0.05).

The detection of Glu could be used as a method of monitoring and evaluating the prognosis of acute traumatic fracture. GSP level could be the important reference of distinguishing Glu rising because of diabetes mellitus or post-traumatic stress response of body.

Surveillance on nasal carriage rate and drug resistance of Staphylococcus aureus among 859 youngsters in Shanghai

NIE Zhiyan, CHEN Xu, CHEN Hong, HAN Lizhong, NI Yuxing

2015, 30(2):  156-159.  DOI: 10.3969/j.issn.1673-8640.2015.02.013

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To survey the nasal carriage rate and drug resistance of Staphylococcus aureus among 859 youngsters in Shanghai.

The nasal specimens were collected from 859 youngsters. The isolates were identified by Gram staining, ornithine decarboxylase test, deoxyribonuclease test, growth test on mannitol salt agar and plasma tube coagulase test. Antimicrobial susceptibility test was performed by disk diffusion method. Staphylococcal protein A (SPA) typing and Panton-Valentine leukocidin (PVL) gene were performed by polymerase chain reaction (PCR).

The nasal carriage rate of Staphylococcus aureus was 7.0% (60 cases), no methicillin-resistant Staphylococcus aureus (MRSA) was detected, and no PVL toxicity factor was found in these isolates. There were 59 isolates with clone diversity, 1 isolate was spa gene negative, and 3 novel spa types were found.

The nasal carriage rate of Staphylococcus aureus is low, and no MRSA is found in these youngsters.

The distribution and drug resistance analysis of Klebsiella pneumoniae in hospital environment

QIU Guangcui, SUN Mingzhong, SHAO Liangrong, JIN Hao, LU Qiang

2015, 30(2):  160-163.  DOI: 10.3969/j.issn.1673-8640.2015.02.014

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To investigate the distribution and drug resistance of Klebsiella pneumoniae in the three-tier medical institutes of Yancheng city, in order to provide the reference for the prevention of nosocomial infection and the rational use of antibiotics.

Klebsiella pneumoniae was determined by VITEK-32 automatic microorganism analyzer, which was produced by France BioMerieux Company. A total of 627 samples were used for Klebsiella pneumoniae determination. Drug sensitivity test of Klebsiella pneumoniae was performed by disk diffusion (K-B ) method. Standard disk diffusion method suggested by the Clinical and Laboratory Standards Institute (CLSI) was employed to conduct the extended-spectrum beta lactamase (ESBL) test.

A total of 213 Klebsiella pneumoniae positive samples were determined from the 627 samples. The positive rate was 33.97%. The highest prevalence of Klebsiella pneumoniae positive samples was observed in the ward of Internal Medicine Department (66.67%), followed by Department of Obstetrics and Gynecology (43.84%) and Treatment Room (43.08%).The resistance rates of Klebsiella pneumoniae to ampicillin and amoxicillin were 96.70% and 99.73%, respectively. A total of 34.27% (73/213) of the determined Klebsiella pneumoniae was ESBL-produced Klebsiella pneumoniae.

The pollution of Klebsiella pneumoniae is common in medical environment with high resistance rate. In order to control the nosocomial infection of Klebsiella pneumoniae, hospital environment disinfection and inspection for Klebsiella pneumoniae pollution are needed to be further strengthened.

Analysis on the pathogenic bacterium distribution and drug resistance among 2 418 cases of blood culture

WU Fangfang, XU Wen, YANG Leyuan

2015, 30(2):  163-166.  DOI: 10.3969/j.issn.1673-8640.2015.02.015

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To understand the blood culture pathogenic bacterium distribution and drug resistance, and to provide the reference for clinical treatment.

By retrospective investigation from January 1, 2011 to December 31, 2013, the bacteria isolated from blood specimens were collected for analysis.

A total of 2 418 blood culture specimens were detected, there were 226 isolates of pathogenic bacteria, and the positive rate was 9.3%, including 143 isolates of gram-negative bacilli (63.3%), 83 isolates of gram-positive cocci (36.7%). The biggest isolation was Escherichia coli (99 isolates, 43.8%), 43 isolates of coagulase-negative Staphylococcus (19.0%) and 20 isolates of Klebsiella pneumoniae (8.8%). Escherichia coli and Klebsiella pneumoniae were sensitive to imipenem, amikacin and piperacillin-tazobactam. Gram-positive coccus is highly sensitive to vancomycin and teicoplanin.

The main isolated blood culture pathogenic bacterium is gram-negative bacillus, and the drug resistance of isolated pathogenic bacterium is severe. The clinical pathogenic bacterium resistance monitoring and the rational use of antimicrobial agents should be strengthened.

Fluorescence in-situ hybridization can provide the early and effective prediction for the prognosis of chronic myeloid leukemia

PENG Youfan, LIU Yang, ZHANG Zhaoxia

2015, 30(2):  167-172.  DOI: 10.3969/j.issn.1673-8640.2015.02.016

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To investigate the early predictive significance of fluorescence in-situ hybridization (FISH) in the long-term prognosis of treating chronic myeloid leukemia(CML) by imatinib(IM).

A total of 60 patients with CML undergoing IM treatment were enrolled, their bone marrow specimens were collected, and they were observed per 3-6 months for 48 months for treatment efficiency by bone marrow cell morphology(BM), FISH, reverse transcription polymerase chain reaction (RT-PCR) and karyotyping(CC).

Applying FISH to detect the patients showed that BCR-ABL positive cell <10% in the 3rd month, the 6th month and the 12th month, which there was high CCR in the 12th month, and the most patients got free-event survival in the 36th and 48th months. Comparing with those whose BCR-ABL positive cell >10%, there was statistical significance (P<0.05). Similarly, applying RT-PCR and CC to detect Bcr-Abl mRNA and positive mitosis phase, there was no statistical difference in the 36th and 48th months(P>0.05), and there was statistical significance only in the 12th month (P<0.05).

FISH is able to provide the early and effective prediction for the long-term prognosis of CML.

Application of protein fingerprinting technology in diagnosing 4 types of leukemia

HUANG Hua, CHEN Linxing, LIN Meice, HUANG Jingyu.

2015, 30(2):  173-176.  DOI: 10.3969/j.issn.1673-8640.2015.02.017

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To establish protein fingerprinting model for the diagnosis of leukemia through determining 4 typical types of leukemia.

A total of 16 acute myeloblastic leukemia M2a samples, 18 chronic myelocytic leukemia samples, 14 acute lymphoblastic leukemia samples and 16 acute myeloblastic leukemia M3 samples were determined by protein fingerprinting technology. The comparison of biomarkers was performed by Biomarker Wizard and Biomarker Patterns 5.0 to establish a diagnosis tree model.

The diagnosis tree model combined with 3 biomarkers ( m/z: 3 376, 8 055 and 9 421) could detect 75% acute myeloblastic leukemia M2a (12/16),100.0% chronic myelocytic leukemia (18/18), 71.4% acute lymphoblastic leukemia (10/14) and 87.5% acute myeloblastic leukemia M3 (12/14).

The protein fingerprinting model established by 3 protein features maybe a novel method for diagnosing 4 types of leukemia.

Evaluation on the performance of Sysmex CS 5100 automatic blood coagulation analyzer

WANG Fang, ZHANG Jun, XU Weijie, YUE Jun, CHEN Xiaoyan, CHEN Jin.

2015, 30(2):  177-180.  DOI: 10.3969/j.issn.1673-8640.2015.02.018

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To evaluate the performance of Sysmex CS 5100 automatic blood coagulation analyzer.

PT, PT(INR), APTT, Fbg, TT, AT, D-dimer (DD) and FDP were measured by Sysmex CS 5100 automatic blood coagulation analyzer. The accuracy, imprecision, Fg linearity (reportable range), reference range, carry-over rate were evaluated, and the correlation with Sysmex CA 7000 automatic blood coagulation analyzer was analyzed.

The accuracy was in the range provided by quality control instructions. The maximum within-run and inter-day coefficients of variation met the requirements of Clia'88. Fbg linear validation test showed that the linearity of Fbg was from 1.071 to 5.355, and the correlation coefficient (r) was 0.9950, which met the specified requirements (r≥0.975).The reference range verification tests showed that the R values of all the tests were >0.9, which proved that the laboratory preset reference range can be applied to the analyzer. The r for several items between CS 5100 and CA 7000 were >0.95, which showed good correlation between the 2 analyzers.

CS 5100 has an excellent accuracy, a good precision, a wide range of Fbg detection as well as a strong anti-interference ability. CS 5100 can totally meet the requirements of clinical laboratories.

Evaluation on the analytical performance of capillary electrophoresis for the determination of HbA_1c

LI Qing, LIU Wenbin, JIN Zhonggan, TANG Liping, WANG Meijuan, OU Yuanzhu, YU Xiaoxuan, JU Yi

2015, 30(2):  181-184.  DOI: 10.3969/j.issn.1673-8640.2015.02.019

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To evaluate the precision, accuracy, linearity and anti-interference ability of capillary electrophoresis (CE) for the determination of hemoglobin A_1c (HbA_1c),and compare it with high performance liquid chromatography (HPLC).

Samples were collected according to each evaluation protocol. Samples were determined by SEBIA CAPILLARYS^TM 2 Flex Piercing HbA_1c analyzer (CE method), ARKRAY HA-8160 automatic HbA_1c analyzer (HPLC) and IFCC reference method. Statistical analysis and linear regression analysis were performed by SPSS 19.0.

The impression at common HbA_1c levels (4.91%-9.67%) was <2%. The CE method and IFCC reference method showed good correlation (R²=0.993). The linearity was good (R²= 0.995) from HbA_1c 5.0% to 17.6%. The results were not significantly influenced by carbamylated hemoglobin, labile HbA_1c, bilirubin, chylomicron and so on (absolute bias ≤0.5%). SEBIA and HA-8160 showed good correlation (R²=0.993).

The precision, accuracy, linearity and anti-interference ability of SEBIA can meet the requirement of routine HbA_1c determination in clinical laboratories.

Expressions and significance of gp96 and Mcl-1 in liver cirrhosis and hepatocellular carcinoma tissues

LI Feng, WEI Qun, HUANG Dongfeng, ZHANG Hon

2015, 30(2):  185-190.  DOI: 10.3969/j.issn.1673-8640.2015.02.020

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To investigate the expressions and clinical pathological significance of gp96 and myeloid cell leukemia-1(Mcl-1) in liver cirrhosis and hepatocellular carcinoma tissues to study preliminarily their relationships with the genesis and progression development of liver cirrhosis and hepatocellular carcinoma.

The expressions of gp96 and Mcl-1 were detected respectively by ENVISION immunohistochemistry in 19 liver cirrhosis tissues (liver cirrhosis tissues beside hepatocellular carcinoma), 32 hepatocellular carcinoma tissues and 21 control tissues (non-liver cirrhosis tissues beside hepatocellular carcinoma). Their relationships of expressions with clinical pathological characteristics of hepatocellular carcinoma were investigated. The expressions of gp96 and Mcl-1 of liver cirrhosis tissues beside hepatocellular carcinoma, hepatocellular carcinoma tissues and non-liver cirrhosis tissues beside hepatocellular carcinoma from one patient were detected, and the positive expression rates were compared.

The positive expression rate of gp96 increased gradually from control, liver cirrhosis to hepatocellular carcinoma groups, which showed significantly different (P<0.05). The positive expression of gp96 had relationships with tumor envelope and TNM staging(P<0.05), and there was no relationship with sex, age, tumor size, serum alpha-fetoprotein (AFP), histological grading and clinical staging (P>0.05). Mcl-1 positive expression in hepatocellular carcinoma and liver cirrhosis groups markedly increased compared to control group, respectively(P<0.05), and there was no statistical significance between liver cirrhosis and hepatocellular carcinoma groups (P>0.05). The positive expression of Mcl-1 had relationships with tumor necrosis and TNM staging(P<0.05). The positive expression rate of gp96 in hepatocellular carcinoma group was significantly higher than those in liver cirrhosis and control groups (P<0.05). The positive expression rate of Mcl-1 in hepatocellular carcinoma group was higher than that in control group, and had no statistical significance with that in liver cirrhosis group (P>0.05).

The overexpressions of gp96 and Mcl-1 are related to the genesis and progression of hepatocellular carcinoma. The gp96 may implicate in the formation and development of liver cirrhosis as well as its subsequent malignant transformation, and may be used as a prognostic biomarker for hepatocellular carcinoma.

Retrospective analysis on blood lead measurement results in eaternal quality assessment organized by the Ministry of Health from 2010 to 2014

YU Xiaogang, YAN Chonghuai

2015, 30(2):  191-194.  DOI: 10.3969/j.issn.1673-8640.2015.02.021

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To analyze retrospectively blood lead measurement results in external quality assessment (EQA) organized by the Ministry of Health from 2010 to 2014, and to investigate the factors which can stabilize and improve the quality of blood lead measurement in order to provide scientific reference for quality control.

A total of 35 EQA results in 7 batches from 2010 to 2014 were analyzed by bias, proficiency testing(PT) score and Z score.

The results of bias met the requirements of the Ministry of Health within the range of quality control. The PT score was 100%. The Z score was <2.0 as satisfaction.

The blood lead measurement system of our laboratory is qualified and suitable for determining large-scale samples. The Z score can eliminate the influence of outliers, thus reflects the contestant's levels more comprehensively and objectively. It can be used as a new EQA indicator.

Complement deficiency and its associated diseases

LI Shengjie, ZHANG Aiping, CAO Wenjun

2015, 30(2):  195-200.  DOI: 10.3969/j.issn.1673-8640.2015.02.022

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Complement system, which has the characteristics of crucial immune responses and cascade reactions, not only has a crucial role in acting as a bridge between innate and adaptive immunities, but also is a part of immune regulatory network, and has important significance in the preservation of immunological homeostasis. Early studies introduced complement component deficiency. The abnormal quality and quantity of complement suggest that complement deficiency is associated with an increasing prevalence of infections and autoimmune diseases. This paper reviews the current common complement deficiencies and their associated diseases, expecting to provide new thoughts for the laboratory diagnosis of diseases.