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    30 July 2020, Volume 35 Issue 7
    Tracking detection analysis of 23 cases of SARS-CoV-2 nucleic acid single target positive samples in Zhengzhou
    NIU Weidong, XU Taibin, SUN Limei, AN Yi, SHI Jun, LI Yi, ZHAO Ruizhen
    2020, 35(7):  633-636.  DOI: 10.3969/j.issn.1673-8640.2020.07.001
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    Objective To perform tracking detection from the first detection of 23 cases of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) nucleic acid single target positive samples in Zhengzhou,and to provide a reference for clinical laboratories. Methods According to the Pneumonia Prevention and Control Plan for Novel Coronavirus Infection,nasal swab and oropharyngeal swab samples from suspected corona virus disease 2019 (COVID-19) patients were collected. The nucleic acid was extracted and detected by real-time fluorescence reverse transcription polymerase chain reaction(RT-PCR). Totally,6 open reading frame lab(ORF1ab) gene and 17 nucleocapsid protein(N) gene single target positive samples were detected. Results Totally,8 samples were positive in the tracking detection,and the positive conversion rate was 34.78%. Conclusions The samples detected single target positive for 2 times should be treated seriously and detected for tracking detection.

    Role of neutrophil/lymphocyte ratio in SARS-CoV-2 infection
    YAN Yufei, JIN Yiming, LIU Mingli, DENG Xuemei, QIN Qin
    2020, 35(7):  637-639.  DOI: 10.3969/j.issn.1673-8640.2020.07.002
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    Objective To investigate the role of neutrophil/lymphocyte ratio(NLR) in severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) infection. Methods A total of 70 patients with SARS-CoV-2 infection were enrolled as infection group,50 patients with fever during the same period were enrolled as fever group,and 50 healthy subjects were enrolled as healthy control group. Blood routine test and C-reactive protein(CRP) determination were performed. Receiver operating characteristic(ROC) curve was used to evaluate the performance of NLR,white blood cell(WBC) count,CRP and the percentage of lymphocyte(LYMPH%) in the diagnosis of SARS-CoV-2 infection. Results NLR and CRP were higher in infection group than those in healthy control group(P<0.001),and the absolute value of lymphocyte(LYMPH#) and LYMPH% were lower than those in healthy control group(P<0.001). NLR and CRP were higher in fever group than those in healthy control group(P<0.05),and LYMPH# and LYMPH% were lower than those in healthy control group(P<0.05). NLR in infection group was higher than that in fever group(P<0.05),and LYMPH#,LYMPH% and CRP in infection and fever groups had no statistical significance(P>0.05). There was no statistical significance for WBC count among the 3 groups(P>0.05). ROC curve analysis showed that the areas under curves(AUC) of NLR,WBC count,CRP and LYMPH% in the diagnosis of SARS-CoV-2 infection were 0.918,0.397,0.695 and 0.785,respectively. Conclusions NLR has good diagnostic performance in the differential diagnosis of SARS-CoV-2 infection.

    Roles of SAA combined determination with CRP and blood routine test in the diagnosis of COVID-19
    FAN Jiubo, SUN Li, HE Jiafu, SHANG Yifei, LU Jiacai, MA Yaping, ZHANG Qin, ZHAO Jianzhong
    2020, 35(7):  640-644.  DOI: 10.3969/j.issn.1673-8640.2020.07.003
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    Objective To investigate the roles of serum amyloid A(SAA)combined determination with C-reactive protein(CRP) and blood routine test in the diagnosis of corona virus disease 2019(COVID-19). Methods Totally,30 patients with COVID-19 in Xiangyang Central Hospital were enrolled as COVID-19 group,and 33 healthy subjects were enrolled as control group. The results of blood routine test and CRP and SAA determinations and their clinical data were collected and analyzed between the 2 groups. Results Compared with control group,body temperature,the percentage of neutrophil(NEUT%),CRP and SAA in COVID-19 group were increased,and the percentage of lymphocyte(LYMPH%) was decreased(P<0.01),but white blood cell(WBC) count had no statistical significance(P>0.05). The positive rate of SAA in the diagnosis of COVID-19 was 93.3%. When the opitmal cut-off value of SAA was 6.43 mg/L,the sensitivity was 100%,and the specificity was 91.9%. The area under receiver operating characteristic(ROC) curve(AUC) was 0.994. The AUC,sensitivity and specificity of SAA for different computed tomography(CT) stages were 0.851,82.4% and 76.9%,respectively,and SAA in different CT stages had statistical significance(P<0.01). Conclusions Hematological indexes,especially SAA,can diagnose COVID-19 rapidly and accurately,which can provide a reference for clinical treatment.

    Morphological and clinical characteristics of near-tetraploid acute lymphocytic leukemia
    ZHANG Xiaohui, TAO Chaoxin, BAI Mingming, CHEN Lei, ZHANG Yuna, DU Huijuan, XING Jiangtao, ZHU Yun
    2020, 35(7):  645-650.  DOI: 10.3969/j.issn.1673-8640.2020.07.004
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    Objective To study the morphological and clinical characteristics of near-tetraploid acute lymphocytic leukemia(NT-ALL). Methods All the clinical data and laboratory examinations(blood smear,bone marrow morphology,immunophenotype and genetic examinations)of 1 NT-ALL patient were collected for comprehensive analysis. Results The patient had acute abdomen. According to bone marrow morphology and immunophenotype,he was diagnosed as B-acute lymphocytic leukemia(B-ALL),and the chromosome was near-tetraploid karyotype(81-103 chromosome). Conclusions Adult NT-ALL is rare. The morphological characteristics are different from those of classic acute lymphocytic leukemia(ALL). The nuclear shape is irregular,the nucleolus is large,the vacuole is large,and the prognosis is poor,which may be related to the abnormal karyotype of near-tetraploid.

    Prediction of platelet implantation by immature platelet ratio in allogeneic stem cell transplantation
    LI Yong, SHI Yuanyuan, ZHU Guoqing, YANG Jiayu, XIA Yonghui, RU Yongxin, YANG Donglin
    2020, 35(7):  651-654.  DOI: 10.3969/j.issn.1673-8640.2020.07.005
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    Objective To study the prediction role of immature platelet ratio(IPF) for platelet(PLT) implantation during allogeneic stem cell transplantation(allo-SCT). Methods A total of 35 allo-SCT cases were enrolled. The PLT count,IPF and the absolute value of immature platelet(A-IPF) in peripheral blood were determined. These parameters were used to predict the success time of PLT implantation during allo-SCT. Results IPF was 1-3 d earlier than PLT count in predicting PLT implantation,and A-IPF could play a good complementary role. Conclusions IPF and A-IPF can predict the time of PLT implantation earlier,which can be used as new parameters for the clinical evaluation of PLT implantation after allo-SCT.

    Level changes of lymphocyte subsets and platelet in peripheral blood of preoperative and postoperative patients with pancreatic cancer
    CHENG Juan, GUAN Shihe, CHEN Liwen, ZHANG Jingjun, CHEN Shuying
    2020, 35(7):  655-659.  DOI: 10.3969/j.issn.1673-8640.2020.07.006
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    Objective To determine the levels of lymphocyte subsets and platelet(PLT) in peripheral blood of preoperative and postoperative patients with pancreatic cancer,and to investigate the correlation between these indicators and clinicopathologic parameters of pancreatic cancer patients. Methods A total of 66 patients with pancreatic cancer were enrolled,and 64 healthy subjects were enrolled as control group. The expression levels of lymphocyte subsets [CD3+T cells,CD3+CD4+T cells,CD3+CD8+T cells,B cells and natural killer(NK) cells] and the level of PLT were determined. The correlations with clinicopathologic parameters were evaluated. Results Compared with control group,the expression levels of CD3+T cells and CD3+CD4+T cells were decreased in preoperative pancreatic cancer group(P<0.05),and the expression levels of CD3+CD8+T cells and PLT were increased(P<0.05). The expression levels of CD3+T cells and CD3+CD4+T cells in postoperative group were higher than those in preoperative group(P<0.05),and the expression levels of PLT were lower than those in preoperative group(P<0.05). There was no statistical significance in the expression levels of CD3+T cells,CD3+CD4+T cells,CD3+CD8+T cells,B cells,NK cells and PLT between postoperative pancreatic cancer group and control group(P>0.05). There was no correlation between the expression levels of CD3+T cells,CD3+CD4+T cells,CD3+CD8+T cells,B cells,NK cells and PLT in preoperative peripheral blood and some clinicopathologic parameters(age,tumor size,tumor site,nerve infiltration and the degree of differentiation)(P>0.05). The expression levels of CD3+CD4+T cells and PLT had correlations with different tissue grades and with or without lymph node metastasis(P<0.05). Preoperative peripheral blood level of CD3+CD4+T cells was negatively correlated with PLT(r=-0.44,P<0.01). Conclusions The cellular immune function of preoperative pancreatic cancer patients is inhibited,and the cellular immune function can be restored after operative treatment. Lymphocyte immunologic abnormalities are associated with PLT and are associated with tissue grading and lymph node metastasis in pancreatic cancer.

    Application of mean neutrophil volume in severe acute pancreatitis with bacteremia
    LIN Han, SONG Ying, CAI Qi, GUO Ping, CHEN Liting
    2020, 35(7):  660-662.  DOI: 10.3969/j.issn.1673-8640.2020.07.007
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    Objective To investigate the role of mean neutrophil volume(MNV) in severe acute pancreatitis(SAP) with bacteremia. Methods A total of 48 patients with SAP were enolled. Peripheral blood samples were collected at admission. White blood cell(WBC) count,the absolute value of neutrophil(NEUT#),MNV,C-reactive protein(CRP) and procalcitonin(PCT) determinations and blood culturing were performed. According to blood culturing results,the patients were classified into 2 groups,SAP with bacteremia group and SAP without bacteremia group. The efficiency of each parameter was analyzed by receiver operating characteristic(ROC) curve. Results CRP and MNV in the 2 groups had statistical significance(P<0.05). There was no statistical significance in WBC count,NEUT# and PCT between the 2 groups(P>0.05). ROC curve analysis showed that the area under curve(AUC) of MNV was 0.775,the sensitivity was 84.6%,the specificity was 66.7%,and the optimal cut-off value was 160.5. The AUC of CRP was 0.708,the sensitivity was 92.3%,the specificity was 48.1%,and the optimal cut-off value was 162.5 mg/L. Conclusions MNV may be used in the auxiliary diagnosis of SAP with bacteremia.

    Activity of pseudolaric acid B combined with fluconazole against Candida albican in vitro
    LI Zhen, YIN Hongmei, CHEN Weiqin, XUE Yingjun, HU Jun, ZHU Bin, HU Xiaobo
    2020, 35(7):  663-667.  DOI: 10.3969/j.issn.1673-8640.2020.07.008
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    Objective To evaluate the activity of pseudolaric acid B(PAB) combined with fluconazole against Candida albican in vitro,and to provide a reference for antifungal treatment. Methods Totally,33 isolates of fluconazole-susceptible and fluconazole-resistant Candida albican were collected. The minimum inhibitory concentrations(MIC) of PAB alone and in combination with fluconazole against Candida albican were determined by microdilution method and chequerboard microdilution method,respectively. The drug interactions were analyzed by fractional inhibitory concentration index(FICI). Time-kill curves were used to monitor dynamically the antifungal activity of PAB combined with fluconazole. Results When PAB used alone,it had similar antifungal effect on fluconazole-susceptible and fluconazole-resistant Candida albican with MIC 4-32 μg/mL. The combination of PAB and fluconazole increased the drug susceptibility of drug-resistant Candida albican. The MIC of fluconazole was decreased from ≥128 μg/mL to ≤16 μg/mL,and the MIC of PAB was decreased to ≤2 μg/mL. Synergism was observed in all 12 isolates of fluconazole-resistant Candida albican with FICI 0.035-0.188,10(47.62%) isolates of fluconazole-susceptible Candida albican showed synergistic effect(FICI 0.188-0.375),and the other 11 isolates of fluconazole-susceptible Candida albican displayed no interaction. Conclusions PAB can inhibit the growth of Candida albican,and shows synergistic effect when used in combination with fluconazole,especially for fluconazole-resistant Candida albican.

    Rule making of M protein recognition by serum protein electrophoresis and its application in M protein screening among healthy subjects
    XU Shuang, LU Jin, ZHAO Lei, PEI Lin, JIA Mei, RONG Rong
    2020, 35(7):  668-672.  DOI: 10.3969/j.issn.1673-8640.2020.07.009
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    Objective To establish and verify the rule of M protein recognition by serum protein electrophoresis,and to investigate the role of this rule in screening M protein among healthy subjects. Methods According to related literatures,the rule of M protein recognition was established according to the region of M protein in serum protein electrophoresis. Serum protein electrophoresis was carried out in 312 random clinical serum samples. M protein was screened according to the established rule of M protein recognition,which was verified by immunofixation electrophoresis. Serum samples of 1 170 healthy subjects and 236 clinical samples excluding hematology,nephrology,rheumatology and immunology,bone tumor,hepatology and geriatrics were collected. Serum protein electrophoresis was performed on each sample. M protein was screened according to M protein recognition rule. Immunofixation electrophoresis was used to confirm and type suspicious samples. Results In the 312 random clinical serum samples,28 samples,including M peak or suspicious M peak,were screened by serum protein electrophoresis,among which 16(5.12%) samples were confirmed as M protein positive by immunofixation electrophoresis. One case was missed in serum protein electrophoresis,and the missed rate was 0.32%. The positive rate of M protein was 2.14%(25/1 170) in 1 170 healthy subjects. The positive rates of M protein were 2.64% in males and 1.82% in females. The positive rate of M protein in males was higher than that in females. Among the 25 M protein positive samples,the positive rate of IgG type M protein was the highest(60%),followed by IgM type(24%). Among the 236 clinical samples excluding hematology,nephrology,rheumatology and immunology,bone tumor,hepatology and geriatrics,10 cases had M protein,and the positive rate was 4.24%. ConclusionsThe established M protein recognition rule have a value in M protein recognition by serum protein electrophoresis,which is worth to be applied widely.

    Roles of plasma thrombin-activatable fibrinolysis inhibitors and serum homocysteine in the diagnosis of acute cerebral infarction
    WANG Boyu, LIU Lingling, ZHANG Qin, YAO Hanxin
    2020, 35(7):  673-675.  DOI: 10.3969/j.issn.1673-8640.2020.07.010
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    Objective To investigate the roles of plasma thrombin-activatable fibrinolysis inhibitor(TAFI) and serum homocysteine(Hcy) levels in the diagnosis of acute cerebral infarction(ACI). Methods The levels of TAFI and Hcy in 100 patients with ACI(ACI group) and 100 healthy subjects(healthy control group) were determined. Receiver operating characteristic (ROC) curve was used to evaluate the roles of TAFI and Hcy in the diagnosis of ACI. Pearson correlation analysis was used to evaluate the correlation between TAFI and Hcy. Results Compared with healthy control group,the level of TAFI in ACI group was decreased(P<0.05),and the level of Hcy in ACI group was increased(P<0.05). The positive rates of TAFI and Hcy in ACI group were 59% and 65%,respectively,which were higher than those in healthy control group(1% and 5%)(P<0.05). ROC curve analysis showed that areas under curves (AUC) of TAFI,Hcy single and combined determinations were 0.907,0.828 and 0.949,respectively. The optimal cut-off values were 22.22 μg/mL,14.95 μmol/L and 0.507,respectively. The sensitivities were 78.3%,65.2% and 88.0%,and the specificities were 95.0%,100.0% and 97.0%,respectively. Pearson correlation analysis showed that there was no correlation between TAFI and Hcy in ACI group(r=-0.075,P>0.05). Conclusions TAFI and Hcy play roles in the diagnosis of ACI.

    Investigation and analysis on pediatric reference intervals of routine chemistry items in China
    DU Yuxuan, WANG Wei, HE Falin, ZHONG Kun, YUAN Shuai, LIU Jiali, ZHANG Zhixin, WANG Zhiguo
    2020, 35(7):  676-681.  DOI: 10.3969/j.issn.1673-8640.2020.07.011
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    Objective To investigate the lower and upper limit difference of pediatric reference intervals of 13 routine chemistry items. Methods The questionnaires on reference intervals of 13 routine chemistry items were distributed to 359 clinical laboratories participating the external quality assessment(EQA) program of the National Center for Clinical Laboratories. The participating clinical laboratories reported reference intervals,methods,instruments and reagents used in prescribed form. Results The number of clinical laboratories participating in routine chemistry reference intervals was 122(30.9%). The primary source of reference intervals were from National Guide on Clinical Laboratory Procedures and instructions of instrument and reagent manufactures(63.0%-69.3%). Totally,44.5%-50.0% of participants had the verified reference intervals. Only a few clinical laboratories had considered different ages and sex when establishing reference intervals. There was statistical significance for lower and upper limits of urea(Urea),creatinine(Cr),aspartate aminotransferase(AST),gamma-glutamyltransferase(GGT) and lower limit of alanine aminotransferase(ALT)(P<0.05). A total of 35.0% of participants had considered different ages and sex in alkaline phosphatase(ALP)(P<0.01,P<0.05),and there was statistical significance for its lower and upper limits for <1-year-old,1-12-year-old,13-15-year-old and >15-year-old children(P<0.05),and there was no statistical significance in the age intra-group(P>0.05). Conclusions The status of pediatric reference intervals of routine chemistry among clinical laboratories in China is not good. It is required to establish the pediatric reference intervals in China according to different ages and sex.

    Interference of endogenous urea in the determinations of TgAb,TPOAb and TSH by chemiluminescence immunoassay
    LI Lihe, SUN Guozhong, ZHANG Chao
    2020, 35(7):  682-685.  DOI: 10.3969/j.issn.1673-8640.2020.07.012
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    Objective To investigate the interference of endogenous urea in the determinations of thyroglobulin antibody(TgAb),thyroid peroxidase antibody(TPOAb) and thyroid-stimulating hormone(TSH) by chemiluminescence immunoassay(CLIA). Methods CLIA was used to determine the levels of serum TgAb,TPOAb and TSH in 30 patients with renal function failure before and after dialysis. Different levels(20,10,30,40,50 and 60 mmol/L) of urea were added to the mixed sera of healthy subjects,and the changes of TgAb,TPOAb and TSH levels were compared. The 800 U/L urease was added to serum samples from patients with renal function failure before and after dialysis,and the changes of TgAb,TPOAb and TSH levels were compared. Results Serum levels of TgAb,TPOAb and TSH of patients with renal function failure after dialysis were higher than those before dialysis(P<0.001). There was no statistical significance for TPOAb and TgAb in TPOAb<15.0 IU/mL and TgAb<15.0 IU/mL dialysis patients before and after dialysis(P>0.05). With the increasing of urea in the mixed sera of healthy subjects,the levels of TgAb,TPOAb and TSH were gradually decreased. After adding 800 U/L urease,there was no statistical significance in TgAb,TPOAb and TSH levels before and after dialysis in patients with renal function failure(P>0.05). Conclusions High level of endogenous urea can interfere with the determinations of TgAb,TPOAb and TSH by CLIA. The addition of urease can eliminate the interference of endogenous urea.

    Serum procalcitonin and urine endotoxin determinations for guiding CAUTI antimicrobial therapy
    LU Xiufen, HE Zhijun, LIU Qiting, FENG Zhongxin, MA Sheng
    2020, 35(7):  686-690.  DOI: 10.3969/j.issn.1673-8640.2020.07.013
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    Objective To analyze the roles of serum procalcitonin(PCT) and urine endotoxin determinations for guiding catheter-associated urinary tract infection(CAUTI) antimicrobial therapy. Methods Totally,180 patients with CAUTI were enrolled and classified into 3 groups,including no drug usage group(60 cases),Gram negative(G-) bacterial antimicrobial group(60 cases) and Gram positive(G+) bacterial antimicrobial group(60 cases),according to the situation of antimicrobial therapy based on serum PCT and urine endotoxin levels. The bacterial determination,drug resistance,nosocomial infection and treatment,biochemical indicators and urinary tract irritation in the 3 groups before and after treatment were compared. Results The drug usage time,dosage,nosocomial infection control time and hospital staying time in G- bacterial antimicrobial group and G+ bacterial antimicrobial group were higher than those in no drug usage group(P<0.001). The bacterial determination rate and CAUTI-related indicators had no statistical significance among the 3 groups(P>0.05). After treatment,the determination rates for resistant isolates in G- bacterial antimicrobial group and G+ bacterial antimicrobial group were lower than those in no drug usage group(P<0.05). Urinary tract irritation indicators,frequent micturition,urgent urination and dysuria,after treatment among the 3 groups had no statistical significance(P>0.05). Conclusions PCT combined with urine endotoxin determination can guide the rational drug usage for CAUTI patients.

    Correlation between serum thyroid hormones and Cr and Cys C in patients with initial thyroid dysfunction
    ZHANG Ke, TONG Shutao, ZHANG Detai
    2020, 35(7):  691-694.  DOI: 10.3969/j.issn.1673-8640.2020.07.014
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    Objective To investigate the correlation between serum thyroid hormones and creatinine(Cr) and cystatin C(Cys C) in patients with initial thyroid dysfunction. Methods A total of 368 patients with thyroid dysfunction,including 108 cases of subclinical hypothyroidism,59 cases of hypothyroidism,102 cases of subclinical hyperthyroidism and 99 cases of hyperthyroidism,were enrolled. Totally,270 healthy subjects were enrolled as healthy control group. Serum thyroid-stimulating hormone(TSH),free triiodothyronine(FT3),free thyroxine(FT4),Cr and Cys C levels were determined. Spearman correlation analysis was used to correlation evaluation. Results The level of serum Cr in hyperthyroidism group and serum Cys C levels in subclinical hypothyroidism group and hypothyroidism group were lower than those in healthy control group(P<0.01,P<0.05,P<0.01). Serum Cys C level in hyperthyroidism group was higher than that in healthy control group(P<0.01). Cr was negatively correlated with FT3 and FT4r=-0.082 and -0.119,P<0.05),and it was positively correlated with TSH(r=0.202,P<0.01). Cys C was positively correlated with FT3 and FT4r=0.281 and 0.197,P<0.01),and it was negatively correlated with TSH(r=-0.214,P<0.01). Conclusions Thyroid hormones in patients with thyroid dysfunction,especially in patients with hyperthyroidism,have effects on serum Cys C and Cr levels.

    Establishment of reference interval of human leukocyte antigen-B27 in apparent healthy children
    ZHANG Li, GE Liang, ZHANG Lanfang, MA Xiaoli, SUN Linchun
    2020, 35(7):  695-698.  DOI: 10.3969/j.issn.1673-8640.2020.07.015
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    Objective To establish the reference interval of human leukocyte antigen-B27(HLA-B27) in apparent healthy children,and to provide a reference for the diagnosis and differential diagnosis of juvenile idiopathic arthritis(JIA)enthesitis related arthritis(ERA) in children. Methods A total of 460 children without conscious spine,peripheral joints,inflammation around the joints or other lesions were enrolled to establish the reference interval. Another 200 apparent healthy children and 50 ERA children were enrolled to verify the reference interval. Peripheral blood HLA-B27 was determined,and the reference interval was established and verified. Results The HLA-B27 mean fluorescence intensity of 460 children showed a skewed distribution(Z=1.615,P=0.011),which was 100(78-121) expressed as median(M) [percentiles(P2.5-P97.5)]. There was no statistical significance for the mean fluorescence intensity of HLA-B27 with different ages and sex(P>0.05). The percentile method was used to establish a 97.5% reference interval on one side,and the HLA-B27 reference interval was <121. Consistency comparison results showed that the new reference interval and the HLA-B27 reference interval provided by manufacturer(0-147) showed a poor consistency(kappa=0.013,P<0.001). According to the new reference interval,the abnormal rate of HLA-B27 in 200 apparent healthy children was 5%,and the positive rate of HLA-B27 in 50 children with ERA was 78%. According to the reference interval provided by manufacturer,the abnormal rate of HLA-B27 in 200 apparent healthy children was 3%,and the positive rate of HLA-B27 in 50 children with ERA was 26%. There was no statistical significance for the abnormal rate of HLA-B27(χ2=1.41,P>0.05),and the difference of positive rate fo HLA-B27 had statistical significance(χ2=49.18,P<0.001). Conclusions The reference interval of HLA-B27 in peripheral blood of children in Nanjing has been established. The specificity of reference interval for the diagnosis of ERA is not different from the reference interval provided by manufacturer,but it improves the diagnostic sensitivity and decreases the rate of missed diagnosis.

    Consistency evaluation of 4 nucleic acid detection kits for severe acute respiratory syndrome coronavirus 2
    YAN Xinsheng, YANG Huihui, HAO Yexia, HU Yuanping, ZHANG Litao, LIAO Xin, FAN Qingkun, LIU Zejin
    2020, 35(7):  706-709.  DOI: 10.3969/j.issn.1673-8640.2020.07.018
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    Objective To evaluate the consistency of 4 severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) nucleic acid detection kits in the diagnosis of corona virus disease 2019(COVID-19). Methods A total of 204 inpatients diagnosed with COVID-19 in Wuhan Asia General Hospital from February 18,2020 to February 24,2020 were enrolled. The residual samples were used to evaluate the consistency of 4 SARS-CoV-2 nucleic acid detection kits approved for emergency registration. Based on the clinical diagnosis of COVID-19,the positive detection of any of the 4 kits was used as a reference. The sensitivities and specificities of the 4 kits for the detections of ORF1ab gene and N gene were compared. The consistency was analyzed statistically by Kappa consistency analysis and Chi-square test. Results The overall detection rates of the 4 kits were 72.4%,78.0%,59.3% and 72.4%,respectively(P=0.011). The detection rates of A,B,C and D kits for ORF1ab gene were 47.7%,31.8%,68.2% and 83.2%,respectively(P<0.000 1). The detection rates of A,B and C kits for N gene were 79.0%,90.5% and 16.2%,respectively(P<0.000 1). Conclusions There is a certain proportion of false negativity in SARS-CoV-2 nucleic acid detection kits,and there is statistical significance in the performance of nucleic acid detection kits with different brands. The detection rate may be improved using nucleic acid detection kits with different brands combinedly.

    Genetic mutation profiling of solid tumors based on targeted-based next generation sequencing
    CHEN Huijuan, WANG Jing, WANG Aiqin
    2020, 35(7):  710-715.  DOI: 10.3969/j.issn.1673-8640.2020.07.019
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    Objective To detect the genetic mutations of solid tumors with a 50-gene hotspot mutational panel,and to interrogate the genetic mutation profiling of solid tumors. Methods A total of 367 various types of solid tumor samples were collected,and the genetic mutation with a 50-gene hotspot mutational panel based on BES 4000 platform was determined. Results Among the 367 solid tumor samples,64.03% samples had at least one gene mutation. Different genetic mutations in 28 genes were determined,and the most frequency mutated genes were EGFR(32.15%),TP53(15.53%),K-ras(8.17%),c-kit(5.72%),APC(3.00%),PTEN(3.00%),B-raf(2.72%),ERBB2(2.72%) and PIK3CA(2.72%). In non-small cell lung cancer(NSCLC),the most frequency mutated gene was EGFR(40.10%),and EGFR mutation just harbored in NSCLC patients. TP53 mutation was found at NSCLC,colorectal cancer,gastric cancer and ovarian cancer patients,and the mutation frequencies were 13.61%,34.38%,33.30% and 50.00%,respectively. K-ras(42.86%) was the most frequency mutated gene in colorectal cancer patients,and that in gastrointestinal stromal tumor was c-kit(53.13%). Among 235 gene mutation samples,28.97%(68/235) had at least 2-gene comutation,and TP53 comutation rate was the highest as 16.60%(39/325). There was no correlation between whether harbored gene mutation and sex,age. The mutation rate of TP53 in ≥58-year-old patients was higher than that <58-year-old patients(P<0.001). In NSCLC,females(P<0.01) and patients with adenocarcinoma(P<0.001) were prone to carry EGFR mutation,and <58-year-old patients were prone to carry ERBB2 mutation. Conclusions Different genetic mutation profiling is determined in solid tumors,TP53 mutation is found in various types of solid tumors and is correlated with patients' age. Target-based next generation sequencing can identify the genetic mutation profiling in solid tumors and provide a reference for clinical treatment.

    Identification of bacteria directly by MALDI-TOF MS
    WANG Junjie, MA Bing, LI Yi, YAN Wenjuan, WANG Shanmei, MA Qiong, ZHANG Jiangfeng, XU Junhong, YUAN Youhua
    2020, 35(7):  716-720.  DOI: 10.3969/j.issn.1673-8640.2020.07.020
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    Objective To investigate the feasibility of identifying bacteria directly by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS). Methods A total of 443 non-repeated positive blood cultures were collected and identified directly by MALDI-TOF MS. The positive blood cultures were sub-cultivated to single bacterium colony and identified by biochemical BD testing(biochemical identification). The accuracies of direct identification for positive blood cultures by MALDI-TOF MS and biochemical identification as the gold standard were compared. Results After excluding 7 false positive samples,among 443 positive blood cultures,the accuracies of direct identification by MALDI-TOF MS were 92.8% for Gram-negative bacteria,81.9% for Gram-positive bacteria,84.2% for fungi and 66.7% for anaerobic bacteria,and the accuracy for rare bacteria was low(57.1%). Conclusions The common bacteria in positive blood cultures could be identified directly by MALDI-TOF MS,which can provide a reference for the rational use of antibiotics.

    Role of TRFIA for detecting pepsinogen in screening ulcer gastric cancer
    HANG Chen, HUANG Biao, PENG Haixia, WEI Xin, XU Weihong, SHENG Huiming, LI Ningli
    2020, 35(7):  721-725.  DOI: 10.3969/j.issn.1673-8640.2020.07.021
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    Objective To investigate the role of time-resolved fluoroimmunoassay(TRFIA) for detecting plasma pepsinogen(PG) Ⅰ,PGⅡ and pepsinogen Ⅰ/pepsinogen Ⅱ ratio(PGR) in screening ulcer gastric cancer. Methods According to the results of gastroscopy and pathology,547 patients were classified into 72 cases of non-atrophic gastritis,232 cases of non-atrophic gastritis with other pathology,42 cases of atrophic gastritis,82 cases of peptic ulcer,15 cases of intraepithelial neoplasia and 104 cases of gastric cancer [43 cases of ulcer gastric cancer(ulcer gastric cancer group) and 61 cases of other gastric cancers(other gastric cancer group)]. TRFIA and chemiluminescence microparticle immunoassay(CMIA) were used to detect plasma PGⅠ and PGⅡ levels of 447 patients with gastric diseases,and PGR was calculated. Receiver operating characteristic(ROC) curve was used to evaluate the efficiency of PGR in the diagnosis of ulcer gastric cancer. Results The results of PGⅠ and PGⅡ of TRFIA and CMIA were positively correlated(r=0.894 and 0.982,P<0.05). The results of TRFIA for samples with PGⅠ≥240 ng/mL had poor consistency with CMIA. Compared with peptic ulcer group,plasma PGⅠ level and PGR in gastric cancer group were decreased(P<0.05),and PGR in ulcer gastric cancer group was decreased(P<0.05),and plasma PGⅠ level in other gastric cancer group was decreased(P<0.05). Plasma PGⅡ level and PGR of ulcer gastric cancer group were higher than those of other gastric cancer group(P<0.05). The area under curve(AUC) of PGR for the diagnosis of ulcer gastric cancer was 0.711. The optimal cut-off value was 18.20,the sensitivity was 72.1%,and the specificity was 70.8%. The AUC for the diagnosis of gastric cancer was 0.797,the optimal cut-off value was 18.37,the sensitivity was 79.8%,and the specificity was 70.8%. Conclusions PG Ⅰ and PG Ⅱ detections by TRFIA have a good correlation with CMIA. The upper detection limit of TRFIA is better than that of CMIA. PGR has certain diagnostic value for ulcer gastric cancer.

    Development of colloidal gold immunochromatography assay for the rapid detection of Legionella pneumophila
    ZHANG Qiu, LI Jie, WANG Meng, WANG Yi, YANG Bo, HU Zheng
    2020, 35(7):  726-733.  DOI: 10.3969/j.issn.1673-8640.2020.07.022
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    Objective To establish a novel colloidal gold immunochromatography assay for the specific detection of Legionella pneumophila(Lp). Methods Lp-PAL recombinant protein was prepared and purified by genetic engineering,and hybridoma cell lines secreting anti-Lp-peptidoglycan-associated lipoprotein(PAL) recombinant protein monoclonal antibody were screened. The indirect enzyme-linked immunosorbent assay(ELISA) and immunoblotting were used to screen and identify the specificities of Lp-1 and Lp-2 monoclonal antibodies. The specificity of antigen-antibody response was identified using bio-layer interferometry(BLI) technology. A colloidal gold immunochromatography test strip was prepared based on double antibody sandwich principle,and its detection performance(specificity,sensitivity and stability) was initially evaluated. Results Totally,2 hybridoma cell lines that secreted anti-Lp-PAL protein antibodies were screened out,and 2 monoclonal antibodies,Lp-1 and Lp-2,were purified. Indirect ELISA showed that Lp-1 and Lp-2 monoclonal antibodies only positively reacted with Lp,and reacted with 10 other common respiratory pathogens(Streptococcus pneumoniae,Moraxella catarrhalis,Haemophilus influenzae,Mycoplasma pneumoniae,Staphylococus aureus,Klebsiella pneumoniae,Klebsiella oxytoca,Acinetobacter baumannii,Pseudomonas aeruginosa and Proteus mirabilis) had no cross-reactivity. The results of western blotting showed that Lp-1 and Lp-2 monoclonal antibodies had strong reactions with natural PAL protein and Lp-PAL recombinant protein,and had no purposeful reaction bands with Streptococcus pneumoniae,Haemophilus influenzae and Moraxella catarrhalis. The verification results of BLI technology showed that the antigen-antibody interactions were strong,and the dissociation rate was slow. Lp-1 and Lp-2 monoclonal antibodies recognized different epitopes of the same antigen,and only had a strong reaction with Lp. They had no specific binding reaction with 10 other pathogens. The minimum detection limit of the prepared colloidal gold immunochromatography test strip was 1×107 CFU/mL,which specifically reacted with the 4 common serotypes of Lp(Lp14,Lp12,Lp9 and Lp6) and did not react with 10 other pathogens such as Streptococcus pneumoniae and Moraxella catarrhalis. The strips could be stored at 25 ℃ for at least 6 months. Conclusions Lp outer membrane protein PAL,which possesses a high degree of surface antigen accessibility and antigenicity,could be used as a molecular marker for Lp infection. The colloidal gold immunochromatography test strip with the features of rapidity,convenience and sensitivity provides a unique tool for the on-site surveillance and diagnosis of Lp infection.