Table of Content

30 June 2021, Volume 36 Issue 6

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Genotype polymorphism distribution of GPⅠa G873A and C807T in cardio-cerebrovascular disease and diabetes mellitus

JIAO Ruibao, TANG Jibin, ZHOU Jiali, ZHU Juanjuan, CHEN Jizhong

2021, 36(6):  581-585.  DOI: 10.3969/j.issn.1673-8640.2021.06.001

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Objective To investigate the polymorphism distribution of aspirin efficacy related platelet membrane glycoprotein GPⅠa gene G873A and C807T in cardio-cerebrovascular disease and diabetes mellitus. Methods A total of 811 patients from September 2018 to October 2020 in Tongling People's Hospital,including 106 cases of cardiovascular diseases,597 cases of cerebrovascular diseases,108 cases of diabetes mellitus,were enrolled,and the polymorphisms of GPⅠa G873A and C807T were determined by pyrophosphate sequencing method. Results Wild type,heterozygous mutation type and homozygous mutation type of GPⅠa G873A and C807T all synchronously appeared in the same cases,the sum of heterozygous mutation type and homozygous mutation type was 47.17%-53.27%,and the sum of allele type A and type T was 24.53%-31.48%. In diabetes mellitus group,homozygous mutation type of 2 gene loci accounted for 13.89%,which was higher than those in cardiovascular disease group（1.89%） and cerebrovascular disease group（8.21%）,while the percentage of heterozygous mutation type（35.19%） was lower than those in cardiovascular disease group（45.28%） and cerebrovascular disease group（45.06%）,and compared with cardiovascular disease group and cerebrovascular disease group,there was statistical significance for genotype distribution（P<0.01）. There was no statistical significance for allele distributions among the 3 groups（P>0.05）. There was no statistical significance for 2 genetic locus genotypes and allele distributions between males and females among the 3 groups（P>0.05）. The cardiovascular and cerebrovascular disease patients by age were classified into 29-60-year-old,61-70-year-old,71-80-year-old and 81-94-year-old groups,respectively,and 2 genetic locus genotype and allele distributions had no statistical significance（P>0.05）. Conclusions Polymorphism distribution of aspirin efficacy related GPⅠa gene G873A and C807T has no correlation with sex and age,and the mutation phenomenon of the 2 gene loci is relatively common. The mutations exist differences between individuals,which might affect the clinical effect of anti-platelet therapy. The rate of homozygous mutation of the 2 gene loci in diabetes mellitus group is higher than those in cardiovascular and cerebrovascular disease groups.

Application of leukocyte morphological parameters in the diagnosis of influenza A and B

CAI Qi, CHEN Liting, GUO Ping, LIN Han, WAN Yinglei

2021, 36(6):  586-589.  DOI: 10.3969/j.issn.1673-8640.2021.06.002

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Objective To investigate the characteristics of leukocyte morphological parameters（LMP） in diagnosing influenza A and B. Methods A total of 77 patients with influenza A,109 patients with influenza B and 101 healthy subjects were enrolled in Shanghai Jiao Tong University School of Medicine Ruijin Hospital from July 2019 to January 2020 and were determined for blood routine test,C-reactive protein（CRP）,influenza A virus and influenza B virus. Results There were 28 CRP-positive patients and 49 CRP-negative patients in influenza A group. There were 23 CRP-positive patients and 85 CRP-negative patients in influenza B group. CRP and influenza A and B viruses were negative in healthy control group. In patients with influenza A and influenza B and healthy subjects with negative CRP results,the percentage of neutrophil（NEUT%）,the percentage of monocyte（MO%）,monocyte forward scattered light central position（Mon_FSC_CenterPos）,monocyte forward scattered light distribution width（Mon_FSC_Width） and high fluorescence intensity cell percentage（HFC%） had statistical significance among the 3 groups（P<0.01）. Among CRP-positive patients with influenza A and B,HFC% and Mon_FSC_CenterPos had statistical significance in influenza A and B groups（P<0.05）. Conclusions Mon_FSC_CenterPos can be used as a good biomarker in differential diagnosis of influenza A and B.

Correlation between red cell distribution width and prostate cancer

HUANG Xiaofeng, FAN Xueming, YAO Tianyue, YUAN Wenhua, ZHAO Zhiyun, SONG Yunxiao

2021, 36(6):  590-595.  DOI: 10.3969/j.issn.1673-8640.2021.06.003

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Objective To determine the level of red cell distribution width（RDW） in peripheral blood of patients with prostate cancer,and to investigate the correlation between RDW and prostate cancer. Methods Totally,497 patients diagnosed with prostate cancer in Xuhui Central Hospital from January 1,2016 to June 1,2020 were enrolled,and 519 healthy subjects in the same period were enrolled as control group. A total of 100 patients with prostate cancer from January 1,2016 and June 30,2017 were enrolled and followed up for 3 years. Detailed physical examination and blood examination results of 88 patients were collected. The data,such as age,weight,diabetes mellitus,hypertension,smoking history and alcohol consumption history,were collected. RDW levels were determined. The correlation between RDW and prostate cancer was evaluated. The subjects were further classified into 4 subgroups（<60,60-69,70-79 and >80 years old） according to ages. Their RDW levels were compared. Results Compared with control group,RDW level in prostate cancer group was higher than that in control group（P<0.001）. In the age subgroups,the RDW level in prostate cancer group was increased compared with that in control group（P<0.001）. Multivariate binary Logistic regression analysis found that RDW increased [odds ratio（OR）=3.300,95% confidence interval（CI） 2.290-4.753,P<0.001,body mass index increased（OR=1.086,95% CI 1.003-1.177,P=0.042） and t-PSA increased（OR=1.255,95% CI 1.198-1.315,P<0.001）were risk factors for prostate cancer. Increased RDW was also a risk factor for prostate cancer in different age subgroups. High RDW level at baseline [hazard ratio（HR）=7.439,95%CI 4.278-21.088,P<0.001] was a risk factor for 3-year overall survival in prostate cancer patients. Conclusions The level of RDW is increased in patients with prostate cancer,which is a risk factor for the development and prognosis of prostate cancer.

Next-generation sequencing for the diagnosis of Toxoplasma gondii infection in patients with chronic kidney disease

ZHOU Yujing, XU Jingjing, XU Jie, LIU Dongmei, HE Jun

2021, 36(6):  596-599.  DOI: 10.3969/j.issn.1673-8640.2021.06.004

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Objective To investigate the application role of next-generation sequencing（NGS） in the diagnosis of difficult pathogen infection. Methods Totally,2 samples of ascites and 4 samples of peritoneal dialysis fluid were collected from a patient with stage 5 chronic kidney disease complicated with abdominal pain. The morphology of sediments was observed by microscopy directly. Peritoneal dialysis fluid and ascites were cultured using routine method. Toxoplasma IgG and IgM antibodies were detected by chemiluminescence immunoassay. NGS was used to detect the pathogenic genes from peritoneal dialysis fluid and ascites samples. Results The suspected Toxoplasma pseudocapsule was found by microscopy. Microbial culture was showed aseptic growth. Toxoplasma IgG and IgM antibodies were negative reaction. NGS showed there was Toxoplasma gondii infection in the peritoneal dialysis fluid. Conclusions NGS has advantages and application prospects in the diagnosis and treatment guidance of difficult pathogen infection.

Study on the drug resistance and molecular epidemiological characteristics of MRSA in Hulunbuir

SUN Gang, SUN Hui, DU Yandan, LI Yinyan, LI Yingzhi, DONG Zhanzhu, TONG Lijun, PENG Bo

2021, 36(6):  600-603.  DOI: 10.3969/j.issn.1673-8640.2021.06.005

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Objective To determine the main epidemic isolate of methicillin-resistant Staphylococcus aureus（MRSA） in Hulunbuir,to study the correlation between gene types and resistance spectra,and to define the origin,transmission and epidemiological characteristics of MRSA. The results may help to find solutions to control MRSA infection and delay the emergence of vancomycin insensitive isolate. Methods Totally,55 non-duplicate MRSA isolates were collected from 5 hospitals from January 2012 to June 2017 in Hulunbuir. Vitek 2 Compact automatic identification system was used to identify the isolates and to perform antimicrobial susceptibility testing. Multilocus sequence typing（MLST） combined staphylococcal cassette chromosome mec（SCCmec） typing were used to determine the gene types of MRSA. Results A total of 6 types,including 2 un-typed isolates,were detected in the 55 MRSA isolates,and the predominant type was SCCmec type Ⅱ（58.18%）. Totally,10 types,including 2 new types,were identified in MLST,and the predominant type was ST59（50.91%）,followed by ST72（10.91%） and ST239（10.91%）. Combining SCCmec typing and MLST,the ST59-MRSA-SCCmecⅡ was the predominant type（38.18%）. The resistance rate of ST59 to ciprofloxacin and gentamicin was low（3.57%）,and no levofloxacin,moxifloxacin and rifampicin resistant isolates were determind;The resistance rates of ST239 to tetracycline,ciprofloxacin,levofloxacin,moxifloxacin,gentamicin and rifampicin were 83.33%. ST72 isolates were not resistant to other antibiotics except erythromycin and clindamycin. The resistance rate of erythromycin and clindamycin in ST239（66.67%） was lower than those in ST59（85.71%） and ST72（83.33%）. Conclusions The predominant type is ST59,followed by ST72 and ST239. Combining with SCCmec typing,the predominant isolate is ST59-MRSA-SCCmecⅡ in Hulunbuir. ST59 is sensitive to ciprofloxacin,gentamicin,levofloxacin,moxifloxacin and rifampin,while ST239 shows a different resistance spectrum and has a high drug resistance rate.

Analysis of the appropriate levels of blood lipid among healthy pregnant women in Zhengzhou

DAI Yanpeng, YUAN Enwu, LIU Junjie, LI Yushan, WANG Quanxian

2021, 36(6):  604-608.  DOI: 10.3969/j.issn.1673-8640.2021.06.006

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Objective To investigate the changes of blood lipid levels among healthy pregnant women,and to define appropriate blood lipid levels for healthy pregnant women. Methods A total of 2 124 healthy pregnant women were enrolled,which were classified into 3 groups according to gestational weeks,including early pregnancy group（276 cases）,middle pregnancy group（961 cases） and late pregnancy group（887 cases）. All the subjects were classified into 2 groups according to ages,including advanced age group（478 cases） and appropriate age group（1 646 cases）. A total of 712 non-pregnant women were enrolled as control group. The levels of triglyceride（TG）,total cholesterol（TC）,high-density lipoprotein cholesterol（HDL-C）,low-density lipoprotein cholesterol（LDL-C）,apolipoprotein A1（apo A1） and apolipoprotein B（apo B） were determined. The appropriate blood lipid levels for healthy pregnant women were defined by percentile mothod [the 2.5th percentile（P_2.5） and the 97.5th percentile（P_97.5）]. Results Except for TG in early pregnancy group,the other blood lipid parameters in early,middle and late pregnancy groups were higher than those in control group（P<0.05）. All the blood lipid parameters in middle and late pregnancy groups were higher than those in early pregnancy group（P<0.05）. All the blood lipid parameters in late pregnancy group were higher than those in middle pregnancy group（P<0.05）. Gestational week-specific appropriate blood lipid levels of TC,HDL-C,LDL-C,apo A1 and apo B had been defined by percentile method. Gestational week-specific and age-specific appropriate levels of TG had been also defined by percentile method. Conclusions Appropriate blood lipid levels of healthy pregnant women in Zhengzhou have been defined,in order to provide a reference for monitoring blood lipid levels during pregnancy.

Establishment of the reference interval of small and dense low-density lipoprotein cholesterol in apparently healthy adults in Chengdu

LI Jing, QUAN Hui, ZENG Zhirong, WEI Yi

2021, 36(6):  609-614.  DOI: 10.3969/j.issn.1673-8640.2021.06.007

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Objective To establish the reference interval of small and dense low-density lipoprotein cholesterol（sd-LDL-C） in apparently healthy adults in Chengdu,and to analyze the correlation between sd-LDL-C and other blood lipid parameters. Methods A total of 846 apparently healthy adults with normal blood lipids aged 20 to 69 years,including 474 males and 372 females,were enrolled. They were classified according to ages（20-29 years old,30-39 years old,40-49 years old,50-59 years old and 60-69 years old groups） and sex（male and female groups）. Plasma sd-LDL-C levels were determined by peroxidase method,and the levels of total cholesterol（TC）,triglyceride（TG）,high-density lipoprotein cholesterol（HDL-C）,low-density lipoprotein cholesterol（LDL-C）,apolipoprotein A1（apo A1）and apolipoprotein B（apo B） were also determined. The percentile method was used to establish the reference interval. The appropriate levels of blood lipids were indicated by medians（M）（P_2.5-P_97.5）. Results There was statistical significance in plasma sd-LDL-C,TC,TG,HDL-C,LDL-C,apo A1 and apo B levels between male and female groups（P<0.05）. Plasma sd-LDL-C levels in males aged from 20 to 49 years were higher than those in females of the same age group（P<0.05）. Plasma sd-LDL-C level in males aged 50 to 59 years was higher than that in females（P>0.05）. Plasma sd-LDL-C level in males aged 60 to 69 years was lower than that in females（P>0.05）. There was statistical significance in the appropriate levels of sd-LDL-C,TC,TG,LDL-C and apo B among age groups（P<0.05）,while there was no statistical significance in the appropriate levels of HDL-C and apo A1 in male group and HDL-C in female groups（P>0.05）. The reference intervals of sd-LDL-C were 0.19-0.93,0.29-0.98,0.31-1.01,0.26-0.99 and 0.30-1.03 mmol/L for males aged 20 to 29,30 to 39,40 to 49,50 to 59 and 60 to 69 years,respectively,and they were 0.19-0.82,0.23-1.03,0.20-0.96,0.26-1.15 and 0.23-1.25 mmol/L for females,respectively. The sd-LDL-C was positively correlated with TC,TG,LDL-C and apo B（r values were 0.601,0.544,0.758 and 0.757,P<0.05）,and it was negatively correlated with HDL-C and apo A1（r values were -0.256 and -0.123,P<0.05）. Conclusions The reference interval of sd-LDL-C has been established in Chengdu.

Correlation between serum 25（OH）D,specific IgE and VAS score in children with allergic rhinitis

ZHAO Xueling, ZHENG Xin, XU Huamei, SHI Hongyao

2021, 36(6):  615-617.  DOI: 10.3969/j.issn.1673-8640.2021.06.008

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Objective To investigate the relationship between serum 25-hydroxyvitamin D [25（OH）D],specific immunoglobulin E（IgE） and visual analog scale（VAS） score in children with allergic rhinitis. Methods Serum 25（OH）D and specific IgE level were determined in 38 children with AR（AR group）,38 children without AR（non-AR group） and 40 healthy children（healthy control group）,and VAS was evaluated in all children with AR. Receiver operating characteristic（ROC） curve was used to evaluate the role of 25（OH）D in judging moderate or severe AR. Results The level of 25（OH）D in AR group was lower than those in non-AR group and healthy control group（P<0.05）,and the level of specific IgE in AR group was higher than those in non-AR group and healthy control group（P<0.05）. The level of 25（OH）D in mild group was higher than those in moderate and severe group（P<0.05）,and the level of specific IgE was lower than those in moderate and severe group（P<0.05）. Correlation analysis showed that VAS was negatively correlated with 25（OH）D（r=-0.637,P<0.001）,and it had a positive correlation with specific IgE（r=0.654,P<0.001）. ROC curve analysis showed that the area under the curve of 25（OH）D was 0.862,and the optimal cut-off value was 10.13 ng/mL,with the sensitivity of 81.3% and the specificity of 78.6%. Conclusions The level of 25（OH）D and specific IgE have certain value in the auxiliary diagnosis and judgment of the severity of child AR.

Correlation between blood lipid and carotid atherosclerotic plaque in elderly patients with coronary heart disease

WANG Linlin, FAN Jun, YE Chenyu, YING Chunmei

2021, 36(6):  618-622.  DOI: 10.3969/j.issn.1673-8640.2021.06.009

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Objective To investigate the correlation between blood lipid and carotid atherosclerotic plaque in elderly patients with coronary heart disease（CHD）. Methods A total of 138 elderly CHD patients with plaque were enrolled as plaque group,and 138 elderly CHD patients without plaque were enrolled as non-plaque group. The clinical general data were collected. Total cholesterol（TC）,triglyceride（TG）,low-density lipoprotein cholesterol（LDL-C）,high-density lipoprotein cholesterol（HDL-C）,apolipoprotein B（apo B）,apolipoprotein（a）[Lp（a）],small and dense low-density lipoprotein cholesterol（sd-LDL-C） and non-high-density lipoprotein cholesterol（non-HDL-C） were determined,and the ratios of sd-LDL-C/LDL-C and LDL-C/HDL-C were calculated. Carotid atherosclerotic extent was graded as 4 according to plaque index（0~3）. Spearman correlation analysis was used to analyze the correlation between different carotid plaque grades and serum factors. Risk factors for carotid atherosclerotic plaque in elderly patients with CHD were analyzed by multivariate Logistic regression. Results Body mass index（BMI）,the proportion of diabetes mellitus history,the proportion of smoking history,serum levels of sd-LDL-C,Lp（a） and apo B,the ratios of sd-LDL-C/LDL-C and LDL-C/HDL-C in plaque group were higher than those in non-plaque group（P<0.05）,and the other indexes showed no statistical significance between the 2 groups（P>0.05）. Serum sd-LDL-C,Lp（a）,apo B levels and sd-LDL-C/LDL-C ratios of grade 1,2 and 3 increased in turn（P<0.05）. Serum sd-LDL-C,Lp（a）,apo B and sd-LDL-C/LDL-C ratio were positively correlated with the grade of carotid plaque sclerosis（r values were 0.502,0.396,0.475 and 0.415,P<0.001）. Multivariate Logistic regression analysis showed that diabetes mellitus,smoking,the increased levels of sd-LDL-C,Lp（a）,apo B and sd-LDL-C/LDL-C ratio were risk factors for carotid atherosclerotic plaque in elderly patients with CHD [odds ratios（OR） were 2.203,2.147,1.514,1.829,1.412 and 1.376,respectively,P<0.05]. Conclusions Serum sd-LDL-C,Lp（a） and apo B levels and sd-LDL-C/LDL-C and LDL-C/HDL-C ratios are correlated with carotid atherosclerotic plaque in elderly patients with CHD.

Role of betaine on the stability of serum lactate dehydrogenase

YU Xiaoxuan, OU Yuanzhu, TANG Liping, LIU Wenbin, LIN Feiran, GE Danhong, GONG Jingkai, ZHU Yuqing

2021, 36(6):  623-626.  DOI: 10.3969/j.issn.1673-8640.2021.06.010

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Objective To study the role of different concentrations of betaine on serum lactate dehydrogenase（LDH） stability at different storage temperatures（2-8 ℃,-20 ℃）. Methods The 5 mol/L betaine was added to serum samples at different ratios. No betaine was added to the samples in group 1. The ratios of betaine to sample（V∶V） were 9∶1,8∶2,7.5∶2.5 and 6∶4 for the samples in groups 2 to 5,respectively. Betaine concentration was 0.5-1.25 mol/L. All serum samples were determined for total LDH activity and LDH isoenzyme electrophoresis analysis before storage,and the results of this assay were used as baseline values. The total LDH activity and the percentage of isoenzymes were determined after storage at 2-8 ℃ and -20 ℃ for different times,respectively,and the differences from the baseline values were calculated. Results The recoveries of total LDH activity for serum samples in groups 2 to 4 ranged from 98.36% to 101.41%,and 95.16%-103.48% for serum samples in group 5. Among them,1 serum sample had an offset higher than the deviation specified in the Chinese health industry standard WS/T 403—2012（4.0%）. Under 2-8 ℃ storage for 30 d,the total LDH activity of serum samples in group 1 was 52.31%-59.95% of the baseline value. The total LDH activities of serum samples in groups 2 to 4 were 98.54%-101.64% of the baseline value,and the results were stable. The total LDH activity of serum samples in group 1 was 88.78%-91.57% of the baseline value when stored at -20 ℃ for 21 d. The total LDH activities of serum samples in groups 2 to 4 were 56.14%-65.34% of the baseline value. The proportions of LDH1 and LDH2 among LDH isoenzymes were increased,the proportion of LDH3 was close to the baseline value,and the proportions of LDH4 and LDH5 were 0 when stored at -20 ℃ for 30 d. Conclusions At 2-8 ℃,0.5-1.25 mol/L betaine stabilizes total serum LDH activity for at least 30 d. Serum LDH is unstable when stored at -20 ℃.

Establishment of cerebrospinal fluid IL-6 reference interval in non-inflammatory central nervous system diseases

WANG Zipeng, OUYANG Changli, GENG Deqin, WANG Dunjing

2021, 36(6):  627-630.  DOI: 10.3969/j.issn.1673-8640.2021.06.011

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Objective To establish the reference interval for cerebrospinal fluid interleukin-6（IL-6） in non-inflammatory central nervous system diseases. Methods A total of 159 patients who were diagnosed as non-inflammatory central nervous system diseases by lumbar puncture examination were enrolled. Totally,23 patients who underwent lumbar puncture and were diagnosed as non-inflammatory central nervous system diseases and 29 patients who were diagnosed as central nervous system infection were enrolled for the verification of reference interval. Results According to the calculation result of Dixon method,1 outlier was eliminated,and there were 158 valid data. The cerebrospinal fluid IL-6 results of 158 patients with non-inflammatory central nervous system diseases showed a normal distribution（Z=0.06,P=0.20）,and the result was（4.43±1.18） pg/mL. There was no statistical significance in cerebrospinal fluid IL-6 levels between males and females（P>0.05）. After merging the groups,the upper limit of the reference interval was 6.74 pg/mL,and the reference interval was ≤6.74 pg/mL using the method of x+1.96s. The cerebrospinal fluid IL-6 results of 23 patients with non-inflammatory central nervous system diseases were 1.50-6.26 pg/mL,and all the results were within the established reference interval. The results of cerebrospinal fluid IL-6 in 29 patients with central nervous system infection were 8.43-3 406.00 pg/mL,which were higher than the established reference interval. The reference interval was validated. Conclusions The IL-6 reference interval of cerebrospinal fluid for patients with non-inflammatory central nervous system diseases has been initially established,and it can be used in clinical practice.

Roles of pretreatment PLR,NLR and LMR in the prognosis of prostate cancer

LI Junsheng, SONG Yunxiao, CHENG Jie

2021, 36(6):  631-636.  DOI: 10.3969/j.issn.1673-8640.2021.06.012

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Objective To investigate the roles of pretreatment platelet/lymphocyte ratio（PLR）,neutrophil/lymphocyte ratio（NLR） and lymphocyte/monocyte ratio（LMR） in the prognosis of prostate cancer patients. Methods The clinical data of 144 patients with prostate cancer were enrolled. Pretreatment platelet,lymphocyte,monocyte and neutrophil were determined. PLR,NLR and LMR were calculated. Patients diagnosed with prostate cancer were enrolled for a 3-year follow-up. Kaplan-Meier method was used for survival analysis. Cox regression analysis was used to evaluate the correlations between PLR,NLR and LMR with the prognosis of prostate cancer. The correlations between PLR,NLR and LMR with Gleason scores were analyzed by Pearson analysis. Results PLR,NLR and LMR of 144 prostate cancer patients were 4.71（1.21-62.14）,4.77（1.34-57.41） and 2.83（0.91-39.10）,respectively. The subjects were grouped according to the median of the measured values of the 3 indicators. In patients with high PLR and low LMR,the 3-year overall survival rate,progression-free survival rate and tumor-specific survival rate were decreased（P<0.001）. There was no statistical significance in 3-year overall survival rate,progression-free survival rate and tumor-specific survival rate of prostate cancer patients between high and low NLR groups（P>0.05）. Cox regression analysis showed that high PLR,low LMR,age,alcohol history,hypertension history and TNM stage were prognostic factors for 3-year overall survival rate,progression-free survival rate and tumor-specific survival rate in prostate cancer patients（P<0.05）. Pearson analysis showed that there were positive correlations between PLR and NLR with Gleason score（r=0.26 and 0.23,P<0.01） and a negative correlation between LMR and Gleason score（r=-0.41,P<0.000 1）. Conclusions PLR and LMR before treatment can be used as prognostic indicators for patients with prostate cancer,and patients with pretreatment high PLR and low LMR have poor prognosis.

Weak expression of antigen caused by the variation in the regulatory region of ABO allele and cause analysis

GU Yuwei, WANG Chengyun, GU Ping, PAN Qiuhui, WANG Jing, CHEN Guangjie

2021, 36(6):  637-641.  DOI: 10.3969/j.issn.1673-8640.2021.06.013

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Objective To analyze the reason of 1 case of the discrepancy of ABO blood group and find out the mutation gene through carrying out genotyping. Methods Serologic blood group was identified by microcolumn gel method,and antibodies were screened and determined with DG GelConfirm cards,DG GelNeutral cards,DG GelCoombs cards. The enhancer,promoter,exon 1-exon 7 and their adjacent intron region of ABO gene were amplified by polymerase chain reaction（PCR）,the PCR products were directly sequenced to identify the gene mutation. Results The patient's red blood cells showed weak agglutination with anti-A（++）,noagglutination with anti-B. The patient's serum showed strong agglutination with A1 cell（++++）,B cell（++++）,noagglutination with O cells（A_weak）. The direct antiglobulin test,indirect antiglobulin test and antibody screening were all negative. Totally,4 short 43 bp tandem repeat units in enhancer region was 3 more than the normal A allele. Conclusions The 4 short 43 bp tandem repeat units in enhancer region of A allele is the cause of the weak expression of A antigen.

Application of BAC spreader for minimum inhibitory concentration determination in Mycobacterium tuberculosis

YU Chenlei, JIANG Yuan, LI Jing, WANG Lili, CHEN Wei, SHEN Xuhui, ZHANG Yangyi

2021, 36(6):  650-655.  DOI: 10.3969/j.issn.1673-8640.2021.06.016

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Objective To evaluate BAC spreader for minimum inhibitory concentration（MIC） determination in Mycobacterium tuberculosis（MTB）. Methods BAC spreader and glass bead grinding were used respectively to prepare bacterial suspensions of MTB ATCC 27294 standard strain H37Rv and 28 clinical MTB strains,and these bacterial suspensions were inoculated into TREK Sensititre MYCOTB well for drug susceptibility test. The performance of BAC spreader on MTB MIC was assessed by visual observation,acid-fast staining,colonial morphology in MYCOTB well and drug susceptibility test results. The role of BAC spreader for MIC determination in MTB was evaluated. Results The results of visual observation and acid-fast staining showed that the bacterial suspensions of BAC spreader were dispersed. The drug susceptibility test results showed that the consistency rate of the 2 methods was 100%. Conclusions The BAC spreader can improve the automation of MTB bacterial suspension preparation,particular in quantitative test as MIC drug susceptibility test,reducing the quantitative error caused by human factors,which is beneficial to the determination standardization and reproducibility.

Identification of MALDI-TOF MS in determining the bacteria from artificial joint replacement infection

DING Yiwei, HAN Zhihai

2021, 36(6):  656-661.  DOI: 10.3969/j.issn.1673-8640.2021.06.017

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Objective To study the role of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF MS） combined with HB&L system to rapidly determine the bacteria from artificial joint replacement infection. Methods The synovial fluid samples of 104 patients with artificial joint replacement infection were collected and inoculated into blood culture flasks and conventional culture agar plates. Positive blood cultures were directly or routinely identified by MALDI-TOF MS,and the type and time of identification were recorded. Results Both direct and conventional MALDI-TOF MS determined a positive result of 84.8%（84/99）,but only 60.6%（60/99） was isolated by plate culture. The determination rates of Gram-positive and Gram-negative bacteria by HB&L direct MALDI-TOF MS were 86.9% and 91.9%,respectively,which were higher than those by gel direct MALDI-TOF MS（78.3% and 83.8%） and plate culture（70.1% and 67.6%）. Among the positive results of HB&L direct MALDI-TOF MS group,the determination rates of Staphylococcus aureus,Staphylococcus epidermidis,Escherichia coli and Klebsiella pneumoniae were high. The identification time of HB&L direct MALDI-TOF MS was shortened to 5-14 h,which was different from 45.2 h of conventional MALDI-TOF MS or 54.3 h of conventional culture. Conclusions Compared with conventional culture,HB&L direct MALDI-TOF MS shortens the time for microbial identification and has an accuracy rate,which provides a reference for the rapid diagnosis of pathogens in artificial joint replacement.

Fresh blood comparison program in daily internal quality control of hematology analyzers

ZHENG Cuiling, WANG Li, CHENG Yan, CUI Wei

2021, 36(6):  662-666.  DOI: 10.3969/j.issn.1673-8640.2021.06.018

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Objective To establish a fresh blood comparison program so as to ensure the comparable and consistent results of clinical specimens,and to improve the daily internal quality control（IQC） of hematology analyzers. Methods After the completion of daily IQC with supporting quality control products,2 mL fresh blood sample was drawn with test results within reference ranges. A hematology analyzer with good performance was selected as the target,and the remaining instruments were used as comparison instruments. The results tested by the target instrument were used as the target values,while the results tested by the comparison instruments were used as the measured values. Comparing the target values and the measured values,the relative deviations of white blood cell（WBC）,red blood cell（RBC）,hemoglobin（Hb）,hematocrit（HCT） and platelet（PLT） were calculated. The 1/2 of allowable error in the Clinical Laboratory Improvement Amendments of 1988（CLIA'88） was the relative deviation standard between different instruments,while the 1/3 of allowable error was the relative deviation standard of the same hematology analyzer at different times. The relative deviation was evaluated according to the relative deviation standard. Results The relative deviations of the comparison data among the comparison instruments and the reference instrument met the requirements of 1/2 CLIA'88 deviation range,showing the accurate and comparable determination results among the instruments. The relative deviations of the comparison data during the operation of the whole cell analyzers met the requirements of the 1/3 CLIA'88 deviation range,indicating the stable instruments. Conclusions The fresh blood comparison program could not only ensure the comparability of determination results among hematology analyzers and the stability of the same instrument during operation,but also perform convenient,economical and effective IQC on the hematology analyzers.

Establishment of a new algorithm for the reference interval of relative change values and application evaluation

YUE Bo, LI Danjie, TANG Dahai, LIU Manjiao, HE Jialei, JIANG Mengjie, WEI Xiaoqiang

2021, 36(6):  667-673.  DOI: 10.3969/j.issn.1673-8640.2021.06.019

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Objective To establish and apply a new algorithm for the reference interval of relative change values（RCV）. Methods Based on the relationship between the relative change values（Δ_R） and the fluctuation（λ）,a related big datum model was established by Excel 2007,and the distribution characteristics of λ and Δ_R and the relationship with coefficient of variation（CV） were analyzed. Finally,the algorithm of RCV was established. RCV data in previous literature reports were recalculated using this algorithm and compared with traditional algorithms. Results λ was positively skewed,and its logarithm was center-symmetrically distributed. When CV≤9.5%,λ was approximately log-normally distributed. The shape of Δ_R distribution was the same as that of λ,and the position was shifted "1" to the left relative to λ. RCV could be derived from CV through this algorithm. Compared with traditional algorithms,when CV was larger,the difference between the 2 algorithms was significant. Conclusions This study establishes a new algorithm for RCV. Compared with traditional algorithms,this algorithm is more scientific and helpful for the clinical popularization and the application of RCV. This study analyzes the relationship between the fluctuation reference interval,RCV and CV through the algorithm,and these 3 factors could be converted to each other.

Assessment of internal quality control plan for microfluidic chip POCT

GE Danhong, TANG Liping, OU Yuanzhu, YU Xiaoxuan, LIU Wenbin, LIN Feiran, GONG Jingkai, ZHU Yuqing

2021, 36(6):  674-678.  DOI: 10.3969/j.issn.1673-8640.2021.06.020

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Objective To preliminarily investigate the internal quality control （IQC） plan for microfluidic chip point-of-care testing （POCT）. Methods Microfluidic chip POCT analyzer was used to analyze potassium（K）,sodium（Na）,chlorine（Cl） and glucose （Glu） from clinical residual serum samples. The study lasted for 3 months and was divided into 2 phases. In Phase 1,electronic quality control and 2 levels of liquid quality control were carried out each day. In Phase 2,electronic quality control was tested on a daily basis,while the liquid quality control was tested once a week. Totally,3 levels of mixed serum samples for these 4 items were tested throughout the study. The results of electronic quality control,liquid quality control and mixed serum samples were analyzed. Results Each result of electronic quality control passed in 2 phases. The liquid quality control results were within control when they were assessed adopting Westgard sigma rules in different phases. The coefficient of variation （CV）of the mixed serum sample results for 4 items were all <1/3 allowable total error （TEa）. Conclusions When microfluidic chip POCT is used to test K,Na,Cl and Glu,under the condition of stable determination personnel and environment,the same batch of reagents,the use of daily electronic quality control with weekly liquid quality control can ensure the quality of analysis.