Table of Content

30 July 2015, Volume 30 Issue 7

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Orginal Article

Updates for clinical laboratories from K/DOQI 2002 to KDIGO 2012 clinical practice guidelines for chronic kidney disease

GUO Wei, PAN Baishen

2015, 30(7):  663-667.  DOI: 10.3969/j.issn.1673-8640.2015.07.001

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Chronic kidney disease(CKD)has been recognized as global public healthy problem. In 2002, Clinical Practice Guidelines for Chronic Kidney Disease: Evaluation, Classification, and Stratification of the Kidney Disease Out-comes Quality Initiative (K/DOQI) from the National Kidney Foundation (NKF) established the definition and framework for the evaluation and management of CKD. In 2012, Clinical Practice Guidelines for Evalution and Management of Chronic Kidney Disease of the Kidney Disease·Improving Global Outcomes (KDIGO) updated and developed an explicit process of evidence review and appraisal, specifically including recommendations for clinical laboratories. This paper reviews the update progress and focuses on clinical laboratory sections in order to learn from the guidelines as well as apply in clinical practice better.

The development and comparison of estimated glomerular filtration rate equations

WANG Yin, WANG Beili, GUO Wei, PAN Baishen

2015, 30(7):  668-673.  DOI: 10.3969/j.issn.1673-8640.2015.07.002

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The key to diagnose kidney diseases early relies on accurate judgement on glomerular filtration rate (GFR). Some estimated glomerular filtration rate (eGFR) equations have been developed in order to facilitate clinical use of GFR based on such filtration markers as creatinine (Cr) and cystatin C (Cys C). The earliest equation is Cockcroft-Gault (C-G) equation which relies on chronic kidney disease (CKD) patients with a small quantity, and it could be influenced with so many factors. Nowadays, C-G equation is thought to be more fit for estimating endogenous creatinine clearance rate. Modification of diet in renal disease (MDRD) equation developed later also relies on CKD patients with more subjects involved. However, it is not accurate being used in healthy subjects and non-kidney disease patients, which partially was led by the standardization problem of filtration marker determination. After Cr and Cys C assay standardizations are completed, the MDRD equation performance in clinical use is still not quite ideal, because the equation is established upon CKD patients that it performs well in this limited subjects. The chronic kidney disease epidemiology collaboration (CKD-EPI) equation is developed on CKD patients and more healthy subjects as well, thus its clinical use is much better. CKD-EPI equation based on Cr is more accurate than those based on Cr and Cys C. The eGFR equations have simplifed the evaluating process of kidney function, however, they are fitted through mathematic models, so the chosen method of measured GFR(mGFR) directly decides whether the developed eGFR equation would be accurate or not. Filtration marker determination could be influenced by so many factors, therefore the consistence of advancing the standardization of filtration markers is also a guarantee to the accuracy of the equations. Considering of the factors such as race, diet and muscle mass, the accuracy of eGFR equations depends on using in similar patients as the equation developing subjects.

The methods for measuring glomerular filtration rate and their clinical application

YU Qian, WANG Beili, GUO Wei, PAN Baishen

2015, 30(7):  674-679.  DOI: 10.3969/j.issn.1673-8640.2015.07.003

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As the best scaleplate for evaluation on kidney function, the glomerular filtration rate (GFR) plays an important role in daily practice and has a great influence on the optimization of diagnosis and treatment. GFR can be estimated through different formulas based on routine text results or measured directly. Bias may arise in certain situations as well as special populations when using estimated glomerular filtration rate (eGFR),while measured glomerular filtration rate (mGFR) is more reliable. Regarding the different properties of common filtration markers such as inulin,creatinine,iohexol, ^99mtechnetium-diethylene triamine pentaacetic acid (^99mTc-DTPA),⁵¹chromium-ethylene diamine tetraacetic acid and iothalamate, we could measure mGFR by means of blood and urine clearance rates or kidney dynamic imaging, and the results shows different accuracies. Endogenous creatinine clearance rate is widely used in common practice for convenience,while plenty of studies have revealed its remarkable bias of overestimating GFR. Regarding the outstanding performances in accuracy, either urine iohexol or ^99mTc-DTPA clearance rate is better for popularization. As the validation for eGFR, mGFR deserves further work on its application and evaluation.

Applicability of glomerular filtration rate equations in different populations

BIAN Bingxian, ZHOU Yunlan, SHEN Lisong

2015, 30(7):  680-683.  DOI: 10.3969/j.issn.1673-8640.2015.07.004

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Today, estimated glomerular filtration rate (eGFR) equations based on general population are widely used, however, some equations fail to provide the same performance in certain population because of different features. This review aims to discuss the development and comparative performance of eGFR equations in different populations, including Chinese, children, patients with diabetes mellitus, patients with heart failure and elderly. We suggest that it should pay attention to the standardization of creatinine and cystatin C (Cys C) determinations and the differentiation and localization of eGFR equations in clinical study and application.

Performance evaluation of seven eGFR equations for the early diagnosis of CKD

CHEN Guiying, YANG Tao, YING Jun, HE Xiaohua

2015, 30(7):  684-690.  DOI: 10.3969/j.issn.1673-8640.2015.07.005

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To evaluate the performance of chronic kidney disease epidemiology collaboration (CKD-EPI) equations, Chinese estimated glomerular filtration rate (eGFR) investigation collaboration equations and FQ-eGFR equations in the early diagnosis of chronic kidney disease (CKD).

Serum cystatin C(Cys C) and creatinine(Cr) concentrations were detected in 283 inpatients with CKD and 9 307 outpatients, and the eGFR of 3 CKD-EPI equations (Cr-, Cys C- and Cys C-C equations), 2 Chinese eGFR investigation collaboration equations (Cys C- and Cys C-C equations) and 2 FQ-eGFR equations (Cys C- and Cys C-C equations) were calculated as the order of eGFR1-eGFR7. Plasma ^99mtechnetium-diethylene triamine pentacetic acid (^99mTC-DTPA) clearance rate as measured glomerular filtration rate (mGFR) was detected in 283 inpatients with CKD, and was used as a gold standard for evaluating difference, consistency and performance of eGFR. The incidence rate ratio (IRR) of each eGFR predicting CKD was analyzed retrospectively in 9 307 outpatients.

There were significant differences between eGFR2 and eGFR3 with mGFR (P=0.000). The concordance correlation coefficients (ρ_c)of eGFR4, eGFR6 and eGFR7 with mGFR were all significantly higher than that of eGFR1 (P<0.01). For the patients with mGFR<60mL/(min·1.73 m²), the deviation, precision and accuracy of eGFR4, eGFR6 and eGFR7 had quite correspondence(P>0.05), and had significant differences with those of eGFR1(P<0.05). In contrast, the deviation of eGFR4 was smaller [median (M)=0.3 mL/(min·1.73 m²)], the precision of eGFR6 was higher [standard interquartile range (IQR)=7.7 mL/(min·1.73 m²)], and the accuracy of eGFR7 was better (1-P₃₀=13.8%). The incidence rates of predicting CKD by eGFR4, eGFR6 and eGFR7 were higher than that by eGFR1 (P<0.01), and were greatly higher in elderly (>65 years old) compared with in under 65 years old (P=0.000).

CKD-EPI equations could not provide an accurate assessment of kidney function for Chinese adult patients with CKD, whose GFR<60 mL/(min·1.73 m²). In contrast, it would be more appropriate for the screening of diseases and early diagnosing used Cys C- equation, FQ-eGFR Cys C- equation or FQ-eGFR Cys C-C equation of Chinese eGFR investigation collaboration equations, in which it is as far as possible to prevent leak-diagnosis or misdiagnosis when FQ-eGFR Cys C- equation is used for the screening of diseases.

Distribution of 16S rRNA methylase gene of Enterobacter cloacae

TANG Cuilian, WANG Fang, CHEN Fangjun, ZHANG Wenbin

2015, 30(7):  691-693.  DOI: 10.3969/j.issn.1673-8640.2015.07.006

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To know the distribution characteristics of 16S rRNA methylase gene of Enterobacter cloacae being resistant to aminoglycoside antibiotics and the relationship between 16S rRNA methylase gene and aminoglycoside drug resistance in Shaoyang, Hunan.

A total of 241 isolates of Enterobacter cloacae were collected from the Affiliated Hospital of Shaoyang Medical College, the People's Hospital of Xinning County, the People's Hospital of Wugang and the People's Hospital of Shaoyang County from August 2012 to May 2014, and were identified by API20E. The drug sensitivity test in vitro was performed by K-B method. The armA, npmA, rmtA, rmtB, rmtC and rmtD genes were determined by polymerase chain reaction (PCR). The relation between drug resistant gene and resistant phenotypes was analyzed.

In the 241 isolates of Enterobacter cloacae, 200 isolotes of them were resistant to aminoglycoside antibiotics, including 41.9% isolates being resistant to amikacin, 68.5% to gentamicin, 70.1% to tobramycin, and 63.5% to netilmicin. Two kinds of 16S rRNA methylase genes were detected, among which were armA (96 isolates, 39.8%) and rmtB (17 isolates, 7.1%), and no npmA, rmtA, rmtC and rmtD genes were detected.

16S rRNA methylase is closely related with the drug resistance of aminoglycoside antibiotics. Genotypes carried by 16S rRNA methylases are somewhat different in bacterial isolates from different areas, and armA is a main kind of 16S rRNA methylase gene in Shaoyang.

Analysis on the results of calcium and 25-hydroxy vitamin D of children in Panyu, Guangzhou

XIAO Xing, HE Jinhua, LI Yuguang, HAN Zeping, HU Shufen, HUANG Hui

2015, 30(7):  694-696.  DOI: 10.3969/j.issn.1673-8640.2015.07.007

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To analyze statistically the levels of calcium and 25-hydroxy vitamin D[25(OH)D] of children in Panyu, Guangzhou, in order to provide reference for child health care.

The levels of calcium and 25(OH)D were determined by atomic absorption spectrometer and electrochemiluminescence immunoassay from 1 902 children. The children were classified into 5 groups according to age, infancy group(1-12 months, 817 cases), toddler age group(1-2 years, 599 cases), preschool age group(3-6 years, 291 cases), school age group(7-11 years,167 cases) and adolescence group(12-18 years, 28 cases). Based on different age groups, each group was subclassified by male and female. The calcium and 25(OH)D levels were analyzed statistically.

The average level of calcium was (1.72±0.19) mmol/L, and the average level of 25(OH)D was (39.12±11.26) ng/mL, and the levels of calcium and 25(OH)D had downward trend with the increasing of age. The levels of calcium and 25(OH)D in infancy group were higher than those in the other 4 groups(P<0.05). The levels of calcium in toddler age group was higher than those in preschool age group and school age group (P<0.05). The levels of 25(OH)D in toddler age group was higher than those in preschool age group, school age group and adolescence group. The levels of calcium and 25(OH)D in preschool age group were higher than those in school age group(P<0.05). There was no statistical significance for calcium and 25(OH)D levels with different sex (P>0.05). Calcium and 25(OH)D were in positive correlation [correlation coefficient (r)=0.221,P<0.05]. Linear equation was Y_calcium=0.004X_25(OH)D+1.573.

The levels of calcium and 25(OH)D are not high among Panyu children. The related schools and parents should supervise their children with calcium supplement rationly.

Evaluation on the stable dose prediction accuracy of Warfarin anticoagulant therapy by pharmacogenetics among Shanghai patients

ZHUANG Wenfang, CHEN Yanhong, LUO Ruiping, WU Jing, XUAN Binbin, CAO Yanan, YANG Li, SHENG Huiming

2015, 30(7):  697-702.  DOI: 10.3969/j.issn.1673-8640.2015.07.008

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To validate the stable dose prediction accuracy of Warfarin anticoagulant therapy by pharmacogenetics among Shanghai patients with pulmonary embolism and heart fibrillation.

The complete clinical data of 198 patients with stable Warfarin dose were collected. The VKORC1-1173C>T, CYP2C9*2 and CYP2C9*3 genotypes were detected by polymerase chain reaction-high resolution melting technique after whole blood genomic DNA extraction with peripheral blood 1.8 mL of sodium citrate anticoagulation 1:9. The 4 prediction algorithms, including IWPC for mixed population, WEN for Chinese Taiwan population, HUANG for Chinese mainland population and OHNO for Japanese, were evaluated and compared by mean absolute error (MAE) and prediction accuracy.

The MAE of HUANG, OHNO and WEN in 4 prediction algorithms and fixed dose model was <±1.0 mg/d with ideal prediction percentage >40%, with the minimum MAE in HUANG. The ideal prediction algorithm with highest percentage was OHNO.

The Warfarin stable dose algorithms by pharmacogenetics for Asian population combined with basic clinical information can predict the dose of Warfarin among Shanghai with clinical application significance.

The combined determinations of MCV and MCH with hemoglobin electrophoresis for screening common thalassemia

LI Hongping, LI Peipei, ZHANG Xuan, ZHU Shue, LIN Jie, YANG Qingwei, CHEN Weiguang, WU Xiaoyang

2015, 30(7):  703-706.  DOI: 10.3969/j.issn.1673-8640.2015.07.009

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To compare the sensitivities and specificities of the combined determinations of mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) with HbA₂ or only HbA₂ determination for screening common thalassemia.

A total of 275 β-thalassemia gene carriers, 44 αβ-compound-thalassemia gene carriers, 186 α-thalassemia gene carriers and 398 non-thalassemia gene carriers were enrolled. MCV and MCH were determined by XE-2100, and hemoglobin electrophoresis was performed by CAPILLARYS 2 system. GAP-PCR and PCR-reverse blot hybridization were used to detect 3 common α-thalassemia genes and 17 common β-thalassemia gene mutations, respectively. The sensitivities, specificities, positive predictive values (PPV) and negative predictive values (NPV) were compared between hemoglobin electrophoresis and the combined determinations of MCV and MCH with hemoglobin electrophoresis in thalassemia diagnosis.

For screening β-thalassemia gene carriers and αβ-compound-thalassemia gene carriers by hemoglobin electrophoresis, the sensitivity was 99.69%, the specificity was 98.28%, the PPV was 96.96%, and the NPV was 99.83%. For screening α-thalassemia gene carriers by the combined determination of MCV with hemoglobin electrophoresis, the sensitivity was 86.56%, the specificity was 83.26%, the PPV was 57.30%, and the NPV was 95.98%. For screening α-thalassemia gene carriers by the combined determination of MCH with hemoglobin electrophoresis, the sensitivity was 92.47%, the specificity was 81.45%, the PPV was 56.39%, and the NPV was 97.66%.

Hemoglobin electrophoresis can determine above 95% β-thalassemia and αβ-compound-thalassemia gene carriers. The combined determination of MCH with hemoglobin electrophoresis can improve sensitivity in thalassemia screening.

Research on single nucleotide polymorphism with initial anticoagulative effect and maintenance dose of Warfarin

WU Jiong, LI Jun, WU Shengchao, ZHANG Chunyan, SONG Binbin, WANG Beili, GUO Wei, PAN Baishen

2015, 30(7):  707-714.  DOI: 10.3969/j.issn.1673-8640.2015.07.010

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To screen genetic factors influencing Warfarin initial anticoagulative effect and maintenance dose.

A total of 212 Chinese Han mechanical heart valve replacement patients were enrolled as initial anticoagulative effect group. △INR_4-1 [the difference between international normalized ratio (INR) at the end of observation and initial INR] was used as an index to screen genetic factors during 4 d after their operations. The obtained results were needed to be identified whether they were associated with other endpoint events like overanticoagulation (INR>3 and INR>4) and time to first therapeutic INR in 3 d. In the maintenance dose group, 266 Chinese Han outpatients who had been taking Warfarin for over 3 months were enrolled. Warfarin maintenance dose was defined as the dose that led to the patients' INR within the therapeutic range measured at least 7 d, which was used to screen genetic factors.

In the initial anticoagulative effect group, CYP2C9 (rs1057910), VKORC1 (rs9923231) and VKORC1 (rs2292566) were associated with △INR_4-1(P<0.05), and CYP2C9 (rs1057910) and EPHX1(rs2292566) were associated with time to first therapeutic INR (P<0.001, P<0.05). In the maintenance dose group, CYP2C9 (rs1057910) and VKORC1 (rs9923231) were associated with daily and weekly maintenance doses (P<0.05).

CYP2C9 (rs1057910),VKORC1 (rs9923231) and EPHX1 (rs2292566) show strongly association with Warfarin initial anticoagulative effect. CYP2C9 (rs1057910) and VKORC1 (rs9923231) are significant factors for maintenance dose.

Study on Th17, Treg cell and the expression of relative cytokines in psoriasis patients

YANG Tingting, WANG Qing, LI Zhi, PIAO Jun, PIAO Jing'ai

2015, 30(7):  715-719.  DOI: 10.3969/j.issn.1673-8640.2015.07.011

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To investigate the clinical significance of T helper cell 17(Th17), regulatory T (Treg) cell and the expression of relative cytokines in psoriasis patients.

Flow cytometry was used to analyze the percentages of Th17 and Treg cells in 28 patients with mild psoriasis, 25 patients with severe psoriasis and 25 healthy controls. The mRNA expressions of retinoid-related orphan receptor gamma t (RORγt) and forkhead bos p3 (Foxp3) were detected by reverse transcription polymerase chain reaction (PCR). The concentrations of interleukin 17 (IL-17) and transforming growth facfor beta (TGF-β) in plasma of psoriasis patients were detected by enzyme-linked immunosorbent assay (ELISA).

The percentages of Th17,RORγt mRNA and IL-17 in psoriasis patients increased compared with healthy controls,and there was a significantly higher levels in severe group than mild group (P<0.05). The percentage of Treg cells, Foxp3 mRNA and TGF-β in severe group were significantly lower than those in healthy controls and mild group(P<0.05).

The percentages of Th17 and Treg cells and their relative cytokines can be used as markers of psoriasis progression.

Significance comparison of immunofixation electrophoresis and bone marrow cell morphology examination in clinical typing of multiple myeloma

CHONG Huifeng, SUN Yun, WANG Chuanfa, WANG Yaqin, CAO Chunluan, ZHOU Ping

2015, 30(7):  720-722.  DOI: 10.3969/j.issn.1673-8640.2015.07.012

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To investigate the significance of immunofixation electrophoresis (IFE) and bone marrow cell morphology examination in clinical typing of multiple myeloma (MM).

According to the staging criterion of International Staging System (ISS) , MM patients were classified into stageⅠ (18 cases), stage Ⅱ (23 cases) and stage Ⅲ (48 cases). The results of serum IFE and bone marrow cell morphology examination in all patients were compared, and the correlation between all indicators and ISS staging was analyzed.

Immunoglobulin (Ig) G was the main type among the clinical stages. The positive rates of M protein and myeloma cell percentage in stage Ⅱ and Ⅲ were higher than those in stageⅠ(P<0.05). The positive rates of M protein in stage Ⅲ and overall positive rate were significantly higher than those of bone marrow cell morphology (myeloma cell percentage ≥10% as standard) (P<0.05). The difference of M protein and bone marrow cell morphology positive rates between the clinical stages was statistically significant (P<0.05).

The positive rate of M protein and myeloma cells percentage have an increasing trend with the increase of clinical ISS stage in MM patients.

Correlation analysis on the changes of DNT cells and T lymphocyte subsets in the chronic progress of hepatitis B

DU Zhenhua, BAO Buhe, WANG Renjie, REN Dangli, ZHANG Minghua, LIU Jiqin

2015, 30(7):  723-727.  DOI: 10.3969/j.issn.1673-8640.2015.07.013

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To investigate the changes of CD3⁺CD4^-CD8^-(DNT) cells and T lymphocyte subsets in the chronic progress of hepatitis B virus (HBV) intection.

DNT cells and T lymphocyte subsets of peripheral blood were detected by flow cytometry in 136 patients with HBV infection, including 33 cases of asymptomatic carrier(ASC), 28 cases of acute hepatitis B(AHB), 28 cases of mild chronic hepatitis B (CHB), 25 cases of moderate CHB, 22 cases of severe CHB, and 39 healthy controls (HC).

DNT cell percentages in ASC, AHB and HC groups were 5.43%±3.31%, 4.75%±2.71% and 4.82%±3.43%, and there was no statistical significance among the 3 groups (P<0.05). DNT cell percentages in HC and AHB group were lower than that in CHB group (from mild group to severe group, DNT cell percentages were 7.97%±4.12%, 8.41%±4.93% and 11.36%±5.01%, P<0.05). The DNT percentage in ASC group was only lower than that in severe CHB group (P<0.05). The DNT percentage had no statistical significance in severe CHB group with those in mild CHB and moderate CHB groups (P>0.05). There was no statistical significance for DNT cell percentages between mild and moderate CHB groups (P>0.05), and they were significantly lower than that in severe group (P<0.05). There was no statistical significance for T lymphocyte subsets in HC group, AHB group and ASC group (P>0.05). With the development of CHB, the percentages of CD3⁺ and CD3⁺CD4⁺CD8^-(CD4⁺) cells decreased, and the percentages of CD3⁺CD4^-CD8⁺ (CD8⁺) cells increased.

The increasing of peripheral blood DNT cell percentage is associated with the progress of CHB.

Evaluation and clinical application of latex-enhanced immunoturbidimetric assay for the detection of plasma neutrophil gelatinase-associated lipocalin

SUN Li, NIU Guoping

2015, 30(7):  734-739.  DOI: 10.3969/j.issn.1673-8640.2015.07.016

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To evaluate latex-enhanced immunoturbidimetric assay (LEIA) for the detection of plasma neutrophil gelatinase-associated lipocalin (NGAL) and its clinical application.

LEIA was used to determine the concentration of plasma NGAL according to the Clinical and Laboratory Standards Institute (CLSI) standardization evaluation protocal. The imprecision, recovery rate, linear range, anti-interference and stability were assessed. The plasma NGAL levels of 86 patients with type 2 diabetes mellitus (T2DM), including 41 patients with diabetic nephropathy(DN), and 40 healthy subjects (healthy control group) were determined.

The within-run coefficients of variation(CV) for low and high levels of NGAL were 3.76% and 1.79%, respectively. The inter-day CV were 6.62% and 3.45%, respectively. The average recovery rate was 97.3%-104.6%. The linear range was from 0 to 5 000μg/L, and the deviation was < 8%. Comparing with similar foreign kit (particle-enhanced immunoturbidimetric assay), the correlation was high (R²=0.996 6). The system biases of 200 μg/L and 700 μg/L were 3.66 μg/L and 11.79 μg/L, respectively. The results met the requirements of manufacturers. There was no significant interference on the determination of plasma NGAL with total bilirubin≤600 μmol/L, hemoglobin≤10 g/L, vitamin C≤0.6 g/L and triglyceride≤15 mmol/L. The reagent could be stable for 35 d in the instrument under the condition of 2-8 ℃. The plasma levels of NGAL in healthy control group were all in manufacturers' reference range. The level of plasma NGAL had been gradually increasing in healthy control group, T2DM group and DN group with statistical significance (P<0.01). The plasma NGAL were positively correlated with serum cystatin C (Cys C) and creatinine (Cr) (correlation coefficients were 0.58 and 0.43, P<0.01).

The LEIA for plasma NGAL determination has high sensitivity and precision, which is fast and easy to operate, and can be used directly on the automatic biochemical analyzer. The results are accurate and reliable. It is suitable for the clinical application for mass determination.

Study on the diversity of in vitro invasion abilities of Staphylococcus aureus clinical isolates with different genotypes

WANG Junrui, WEI Changmei, TA La, CUI Jinghua, DU Xiaoli, HAN Yanqiu

2015, 30(7):  740-746.  DOI: 10.3969/j.issn.1673-8640.2015.07.017

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To determine the diversity of in vitro invasion abilities of Staphylococcus aureus clinical isolates with different genotypes, and further to investigate the pathogenesis mechanism of Staphylococcus aureus and provide experimental reference for controlling its nosocomial infection and transmission.

Pulse field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to do molecular typing for 43 isolates of Staphylococcus aureus, and antibiotics protection assay and flow cytometry internalization assay were used to determine the diversity of in vitro invasion abilities of Staphylococcus aureus clinical isolates with different genotypes.

A total of 43 isolates of Staphylococcus aureus were classified into 12 PFGE genotypes according to 80% similarity, among which type Ⅰ and type Ⅱ were the predominant types and accounted for 25.6% (11/43)and 34.9%(15/43), respectively. Other non-predominant types accounted for 39.5%(17/43). In vitro antibiotics susceptibility test results showed that type Ⅰ and other non-predominant types were mainly composed of methicillin-sensitive Staphylococcus aureus (MSSA)(85.7%, 24/28), while type Ⅱ were mainly composed of methicillin-resistant Staphylococcus aureus (MRSA)(100.0%, 15/15). MLST revealed that type Ⅱ isolates were all ST-239 type, but type Ⅰ and other types belonged to diverse ST types. Antibiotics protection assay showed that the invasion rates of the 3 isolates with the strongest invasion abilities were 3.27%, 3.15% and 3.09%, respectively. The in vitro invasion abilities of type Ⅰ and type Ⅱ isolates were significantly higher than those of other non-predominant types (P<0.01).

Staphylococcus aureus clinical isolates with different genotypes shows the diversity of in vitro invasion abilities,and the predominant clones revealed stronger in vitro invasion abilities,especially for ST-239 type.The units with higher Staphylococcus aureus isolation rates should pay more attention to this clone and strengthen infection control measures.

Application of MALDI-TOF MS in direct identification of clinical microbiological samples

CHEN Fei, HU Binjie, ZHAO Hu

2015, 30(7):  750-756.  DOI: 10.3969/j.issn.1673-8640.2015.07.019

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Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS),as a newly developed technology in recent years,is an easy,rapid,accurate and cost-effective method for microbial identification in clinical microbiology, compared to conventional phenotypic techniques or molecular biology. Given the accuracy and reliability of MALDI-TOF MS in the identification of microorganisms grown on solid medium,this technology might also be directly applied to some clinical microbiological samples, such as positive blood culture bottles,midstream urine, cerebrospinal fluid and so on. The goal of this review was to discuss the application of MALDI-TOF MS technology in direct identification of clinical microbiological samples, including the principles, protocols, performance and research progress.

Research progress on the bacterial inoculation techniques in clinical microbiology laboratories

WANG Jinghua, GE Ping, CHEN Rong, XU Rong, LIU Xuejie

2015, 30(7):  757-760.  DOI: 10.3969/j.issn.1673-8640.2015.07.020

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In recent years, researches on the bacterial inoculation automation are booming. The commercialized automatic or semi-automatic microbial inoculation instruments are developing, such as, Robobact system, EasySpiral Pro and PREVI Isola. However, the experience of clinical application indicates that no instrument employed is satisfactory, let alone the accurate quantitative inoculation instrument, and they are hard to be popularized in clinical microbiology laboratories. Meanwhile, to develop a new method of accurate quantitative determination and to unify and standardize operating procedure, is the development tendency of quality control. Based on Bernoulli principle and fluid mechanics,a new-type instrument, called the multichannel microbial inoculation instument, is invented by Chinese scholars. It is hopeful to develop a new method for the quantitative determination for microbiology based on automatic technique. In this review, the research and development progress of bacterial inoculation technique in clinical microbiology laboratories are summarized.

Clinical application of female hormone determination in infertility treatment

ZHANG Chunyan, PAN Baishen

2015, 30(7):  761-765.  DOI: 10.3969/j.issn.1673-8640.2015.07.021

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Laboratory determination for female hormones plays a large part in infertility treatment, especially during in vitro fertilization(IVF) cycle. The hormones contain follicle stimulating hormone(FSH), estradiol(E2), anti-Müllerian hormone(AMH), inhibin B(Inh B), progesterone and luteinizing hormone(LH). Serum FSH, E2 and AMH levels are commonly determined for women as a part of ovarian reserve testing. E2 levels are also frequently evaluated in a controlled-ovarian-hyperstimulation(COH) setting to monitor follicular development. In addition to its role in evaluating ovarian reserve, researchers have studied AMH as a predictor of response to gonadotropin stimulation during ovulation induction. Inh B levels are not regarded as a reliable assessment index of ovarian reserve. Poorer responders to gonadotropin generally have lower Inh B levels. Progesterone is a simple determination for ovarian function. The role of progesterone and LH levels in evaluating the success of IVF cycle has been the subject of debate for many years.