Table of Content

30 July 2021, Volume 36 Issue 7

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Role of anti-nuclear antibody-dense fine speckled pattern for diagnosing systemic autoimmune rheumatic disease

YANG Keke, WU Junyuan, CAO Xingjian

2021, 36(7):  691-695.  DOI: 10.3969/j.issn.1673-8640.2021.07.002

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Objective To investigate the role of anti-nuclear antibody（ANA）-dense fine speckled pattern in the diagnosis of systemic autoimmune rheumatic disease（SARD）.Methods A total of 1 683 samples from suspected or diagnosed patients suffering from SARD were determined for ANA by indirect immunofluorescence（IIF） and anti-extractable nuclear antigen antibody（ENA） by immunoblotting test. The identifications of ANA type were performed on samples positive by IIF but negative by immunoblotting test and both positive by IIF and immunoblotting test. The correlation of ANA-dense fine speckled pattern and the occurrence of SARD was evaluated by rank correlation analysis. Results There was no statistical significance between the positive rates by IIF-ANA and immunoblotting test-ENA（P=0.155）. In 72 samples only positive by IIF,the common speckled pattern accounted for 44.44%,and dense fine speckled pattern accounted for 40.28% without statistical significance（P=0.701）. The homogeneous pattern（12.50%） and nucleolus pattern（2.78%） showed statistical significance with dense fine speckled pattern（P=0.001,P<0.001）. There was no correlation between dense fine speckled pattern and SARD（r=-0.300,P=0.667）. Dense fine speckled pattern appeared in none of the 644 samples both positive by IIF-ANA and immunoblotting test-ENA. Conclusions The positive ANA-dense fine speckled pattern may be applied in excluding SARD.

Role of serum anti-Müllerian hormone levels in the evaluation of ovarian reserve in females with antiphospholipid syndrome of childbearing age

WU Junqi, ZHAO Shuo, WANG Limin, LÜ Ying, FU Miao

2021, 36(7):  696-699.  DOI: 10.3969/j.issn.1673-8640.2021.07.003

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Objective To investigate the roles of serum anti-Müllerian hormone（AMH） and inhibin B（INHB） levels in the evaluation of ovarian reserve in females with antiphospholipid syndrome（APS） of childbearing age.Methods Totally,86 female patients with APS（APS group）,among whom 32 were primary antiphospholipid syndrome（PAPS）（PAPS group）,and 54 are secondary antiphospolipid syndrome（SAPS）（SAPS group）,were enrolled. A total of 52 female patients suffered from autoimmune diseases were enrolled as disease control group,and 50 healthy childbearing females were enrolled as healthy control group. All blood samples were collected within 2-5 d of a new menstrual cycle,which were determined for serum AMH,INHB,luteinizing hormone（LH）,follicle stimulating hormone（FSH） and estradiol（E2） levels,and antral follicle count（AFC） was conducted simultaneously by transvaginal sonography. The correlations were evaluated by Pearson correlation analysis.Results The levels of AMH,INHB,E2 and AFC in APS group and disease control group were lower than those in healthy control group（P<0.01）. AMH in SAPS group had lower results than that in PAPS group（P<0.05）. The ratio of AMH<0.9 ng/mL and the ratio of AMH<0.5 ng/mL in APS group and disease control group were higher than those in healthy control group（P<0.01）,but there was no statistical significance between APS group and disease control group（P>0.05）. In APS group,AMH was positively correlated with INHB and AFC（r values were 0.85 and 0.74,P<0.01）.Conclusions Serum AMH plays a role in the evaluation of ovarian reserve in female APS patients of childbearing age.

Consistency of the results of anti-HCV antibody gray zone samples among different determination systems

LU Yinhua, ZHU Yuqing, ZHAO Xiaojun, CAO Danru, ZHU Lingfeng, GU Zhidong

2021, 36(7):  700-704.  DOI: 10.3969/j.issn.1673-8640.2021.07.004

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Objective To investigate the consistency and correlation between 4 domestic determination systems and ARCHITECT i2000SR automatic immunoassay analyzer with supporting reagents（i2000SR system） for determining anti-hepatitis C virus（HCV） antibody gray zone samples. Methods Totally,4 domestic determination systems [2 systems using enzyme-linked immunosorbent assay（ELISA）,1 system using time-resolved fluorescence immunoassay（TRFIA） and 1 system using chemiluminescence immunoassay（CLIA）] were used to determine different concentrations of anti-HCV antibody standard products and to evaluate the linearity of each determination system and imprecision [coefficient of variation（CV）]. The 4 domestic determination systems and the i2000SR system were used to determine 120 samples of anti-HCV antibody gray zone,which were confirmed by recombinant immunoblotting assay（RIBA） to evaluate the correlation and consistency of each determination system.Results For the 4 domestic determination systems,the within-run CV were 17.6%-25.1%,0.8%-26.7%,7.8%-36.2% and 0.7%-20.2% for the standard products with different concentrations,respectively. The total CV were 15.9%-25.3%,1.0%-23.5%,7.7%-34.8% and 3.4%-18.9%,respectively. The linear correlation equations were Y=0.634X+0.973,Y=2.495X+0.010,Y=1.348X+0.896 and Y=2.510X-0.214,respectively. The S/CO value of the i2000SR system was 1.00-3.00 samples,and the determination consistency rate of the 4 domestic determination systems was 34.2%-55.2%. The determination consistency rates of the 4 domestic determination systems with S/CO values of 3.01-6.00 samples were 57.5%-80.0%. The determination consistency rates of the 4 domestic determination systems with S/CO values of 6.01-9.00 samples were >90%. The number of samples with negative results from the 4 domestic determination systems was 23（19.2%）,and the S/CO value $\bar{x}±3$s was 2.80±4.35. The number of samples with positive results from the 4 domestic determination systems was 71（59.1%）,and the S/CO value $\bar{x}±3$s was 5.13±7.84. The correlation coefficients（r）between 2 domestic reagents were 0.712 5,0.760 0,0.837 0,0.702 2,0.769 4 and 0.736 4,respectively. The Kappa values among the domestic reagents were 0.666 3,0.598 5,0.742 6,0.759 3,0.879 0 and 0.664 8,respectively. Conclusions For the gray zone samples（S/CO value of 1.00-9.41）,it is recommended that at least 2 reagents or highly specific reagents should be used for retesting. For low-concentration samples or cut-off samples,the possible inconsistencies between different determination systems should be fully considered,and a reasonable gray zone range should be formulated.

Role of p2PSA,p2PSA% and PHI in the diagnosis of prostate cancer

QUAN Heng, ZHU Jing, LIAO Huanjin, WU Jun

2021, 36(7):  705-709.  DOI: 10.3969/j.issn.1673-8640.2021.07.005

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Objective To investigate the diagnostic value of prostate-specific antigen isoform 2（p2PSA）,prostate-specific antigen isoform 2 percentage（p2PSA%） and prostate health index（PHI） in the identification of benign and malignant prostate diseases.Methods A total of 250 male patients who underwent transrectal ultrasound-guided prostate biopsy were enrolled and classified into benign prostate disease group（145 cases） and prostate cancer group（105 cases） based on the pathological results,and they were classified into ≤65 years old and >65 years old groups according to ages. Total prostate-specific antigen（t-PSA）,free prostate-specific antigen（f-PSA） and p2PSA were determined in serum samples from all the patients,and free prostate-specific antigen percentage（f-PSA%）,p2PSA% and PHI were calculated. The efficiency of each index in the diagnosis of prostate cancer was evaluated by receiver operating characteristic（ROC） curve. Results Compared with benign prostate disease group,the t-PSA,p2PSA,p2PSA% and PHI of prostate cancer group were increased（P<0.01,P<0.001）,and f-PSA% was decreased（P<0.001）. There was no statistical significance in f-PSA between the 2 groups（P>0.05）. ROC curve analysis results showed that,in ≤65 years old patients,the areas under curves（AUC） of t-PSA,f-PSA,f-PSA %,p2PSA,p2PSA% and PHI in the diagnosis of prostate cancer were 0.687,0.500,0.792,0.815,0.795 and 0.892,respectively. In >65 years old patients,the AUC of t-PSA,f-PSA,f-PSA %,p2PSA,p2PSA% and PHI in the diagnosis of prostate cancer were 0.748,0.542,0.900,0.780,0.843 and 0.929,respectively.Conclusions The diagnostic efficiency of prostate cancer markers differs in different ages. Prostate cancer markers,PHI,p2PSA and p2PSA%,can improve the differential diagnosis of benign and malignant prostate diseases.

Investigation on the process of serological screening for syphilis in pregnant women

QI Jiehua, ZHANG Chunli, WU Shoule, HUAN Yu, CHEN Yu, LE Jiangman, CAI Xushan

2021, 36(7):  710-713.  DOI: 10.3969/j.issn.1673-8640.2021.07.006

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Objective To investigate preliminarily the process of serological screening for syphilis in pregnant women. Methods A total of 548 serum samples of pregnant women with anti-Treponema pallidum（TP）antibody S/CO ratio ≥1.0 screened by chemiluminescence immunoassay（CLIA） were collected and were retested by chemiluminescence microparticle immunoassay（CMIA）,toluidine red unheated serum test（TRUST） and western blotting. The positive predictive values of single method and combined method were analyzed respectively based on the western blotting results,and the serological screening process for syphilis in pregnant women was established. Results There were 548 samples with anti-TP antibody S/CO ratio ≥1.0 determined by CLIA,among which 494 cases were positive, 11 cases were indeterminate（eliminated）, and 43 cases were negative determined by western blotting. Totally,295 samples were positive,and 253 samples were negative determined by TRUST. Totally, 490 samples were positive with S/CO ratio ≥1.0, and 58 samples were negative with S/CO ratio <1.0 determined by CMIA. The results of western blotting were used as gold standard, the positive predictive values of CLIA/TRUST/CMIA（+/-/-）, CLIA/TRUST（+/-）, CLIA/TRUST/CMIA（+/-/+）, CLIA/TRUST（+/+） and CLIA/TRUST/CMIA（+/+/+）were 22.4%, 82.2%, 97.4%, 100.0% and 100.0%, respectively. According to the receiver operating characteristic（ROC） curve, the optimal anti-TP antibody S/CO ratio of CMIA was 2.31 among CLIA/TRUST/CMIA（+/-/+） samples,the sensitivity was 94.7%, and the specificity was 100.0%. The serological screening process for syphilis was established based on the above results：prescreening by CLIA,the negative result can be issued directly,and the positive result should be tested by TRUST;the positive result can be issued,but the negative result should be retested by CMIA;issue the positive result when the S/CO≥2.31,retested by western blotting when the result was negative or S/CO<2.31, and issue the report according to western blotting eventually. Conclusions The serological screening process for syphilis has been established through the combination of 4 syphilis serological methods, which can improve the accuracy of syphilis diagnosis in pregnant women while reducing reexamination by western blotting.

21-Gene recurrence score in mucinous breast cancer

LIN Jiafei, CHEN Xiaosong, WU Beiying, CAI Gang, LIN Lin

2021, 36(7):  714-718.  DOI: 10.3969/j.issn.1673-8640.2021.07.007

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Objective To investigate the clinical value of 21-gene recurrence score（RS） in mucinous breast cancer（MBC）. Methods Totally,96 MBC patients with estrogen receptor（ER） positive and human epidermal growth factor receptor 2（HER-2） negative who underwent surgical treatment were enrolled,and the clinical data of all the patients were collected. RNA was extracted from the tumor paraffin specimens of patients. Real-time fluorescence quantitative polymerase chain reaction（PCR） was used to determine the expression of 21 genes,and 21-gene RS was calculated. According to the scores,all the patients were classified into low-risk group（<18 points）,intermediate-risk group（18-30 points）and high-risk group（>30 points）. Logistic regression was used to analyze the influence factors of 21-gene RS. Kaplan-Meier curve was used to assess disease-free survival（DFS）.Results Among the 96 patients with breast cancer,there were 23 patients（24.0%） in the low-risk group,63 patients（65.6%） in the intermediate-risk group and 10 patients（10.4%） in the high-risk group. There was statistical significance in progestin receptor（PR） expression,molecular typing and tumor size among the low-risk group,intermediate-risk group and high-risk group（P<0.05）. There was no statistical significance in age,menopausal time,staging and Ki67 expression（P>0.05）. Multivariate analysis demonstrated that PR status was independently associated with high risk based on 21-gene RS [odds ratio（OR）=7.271,95% confidence interval（CI） 0.728-3.240]. Univariate analysis found that the choice of chemotherapy was related to Ki67 expression,molecular typing,PR expression and 21-gene RS（P<0.05）．There was no correlation in age,tumor size,menopausal time,operation method and staging（P>0.05）. Multivariate analysis showed that Ki67 expression and 21-gene RS independently influenced the use of chemotherapy（P<0.05）. No statistical significance in DFS was found among MBC patients in different 21-gene RS subgroups（P>0.05）.Conclusions 21-Gene RS is related to PR status in MBC patients. 21-Gene RS does independently affect the choice of chemotherapy. However,21-gene RS is not able to predict DFS in MBC patients.

Comparison on the determination rates of respiratory virus antigens between nasopharyngeal swabs and oropharyngeal swabs

SHAN Jiani, WANG Tianlin, ZHAO Shuiai, FAN Liyun, LIU Lei

2021, 36(7):  719-721.  DOI: 10.3969/j.issn.1673-8640.2021.07.008

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Objective To compare the positive determination rates of common respiratory virus antigens in nasopharyngeal swabs and oropharyngeal swabs,and to provide a reference for clinical selection of sample types.Methods The oropharyngeal swab samples and nasopharyngeal swab samples were collected from 199 children with upper respiratory tract infection. According to random principle,the sequence of samples collected from different patients was determined,and 2 samples were submitted for determination at the same time. Influenza virus（Flu）A,FluB,respiratory syncytial virus（RSV） and adenovirus（ADV） antigens were determined by colloidal gold immunoassay. Results Of the 199 children with upper respiratory tract infection,the total positive rates of FluA,FluB,RSV and ADV were 31.2%,10.6%,6.0%,5.5% and 52.3% for nasopharyngeal swabs and 20.6%,7.0%,5.0%,5.0% and 37.6% for oropharyngeal swabs,respectively. The positive rates of FluA,FluB and 4 viruses were higher in nasopharyngeal swabs than those in oropharyngeal swabs（P<0.05）,while the positive rates of RSV and ADV between the 2 kinds of samples showed no statistical significance（P>0.05）. Nasopharyngeal swabs were positive,and oropharyngeal swabs were negative in 40 children. Nasopharyngeal swabs were negative,and oropharyngeal swabs were positive in 11 children. Nasopharyngeal swab results were used as standard,the sensitivities of oropharyngeal swabs to FluA,FluB,RSV and ADV were 59.7%,61.9%,75.0% and 45.5%,respectively,and the specificities were >97%.Conclusions Combined with the convenience of operation,nasopharyngeal swabs should be selected for respiratory virus determination in children when taking a single pharyngeal swab sample. Since nasopharyngeal swabs are negative,and oropharyngeal swabs are positive in a few cases,samples from both nasopharyngeal and oropharyngeal swabs may be collected for highly suspected cases in clinical practice in order to improve the positive rate.

Etiological analysis of influenza surveillance in Chizhou from 2018 to 2019

LIU Shiyun, TAN Ping

2021, 36(7):  722-725.  DOI: 10.3969/j.issn.1673-8640.2021.07.009

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Objective To analyze the etiological characteristics of influenza in Chizhou from 2018 to 2019,and to provide a reference for the prevention and treatment of influenza. Methods Nucleic acid in the samples of pharyngeal swabs from influenza-like cases was determined by real-time fluorescence quantitative polymerase chain reaction （PCR）. Results The determination rates of influenza virus were 30.09% in 2018 and 28.92% in 2019（P>0.05）. The pandemic was characterized by a mixture of influenza A and B viruses. There was no statistical significance for the determination rate of influenza virus between males and females（P>0.05）. The determination rate in 5-14-year-old group（37.03%） was the highest. The virus determination rate of the outbreak was higher than that of the sentinel monitoring（P<0.01）.Conclusions The prevention and control of influenza in Chizhou focuses on a mixture of influenza A and B viruses,and the key population is teenagers and children,especially primary and secondary school students. The surveillance of influenza etiology should be strengthened,and the pandemic situation should be monitored.

Correlation between nuclear magnetic resonance spectroscopy and traditional biochemical determination methods analyzer for determining blood lipid

PENG Yanjun, HAN Xuejing, TANG Hongxia, CHEN Junmeng, JIA Kegang

2021, 36(7):  733-737.  DOI: 10.3969/j.issn.1673-8640.2021.07.012

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Objective To investigate the relationship of nuclear magnetic resonance spectroscopy（NMR） and traditional biochemical determination methods for determining small and dense low-density lipoprotein cholesterol（sd-LDL-C）,low-density lipoprotein cholesterol（LDL-C）,high-density lipoprotein cholesterol（HDL-C）,total cholesterol（TC） and triglyceride（TG）,and to analyze the level of sd-LDL-C in patients with inconsistent LDL-C and low-density lipoprotein particle（LDL-P）. Methods NMR and traditional biochemical determination methods（biochemical analyzer） were used to determine sd-LDL-C,LDL-C,HDL-C,TG and TC levels in 54 patients,and NMR was used to determine LDL-P level. Pearson correlation analysis was used to evaluate the correlation of the 2 methods. Results The r ² values of traditional biochemical determination methods and NMR for sd-LDL-C,LDL-C,HDL-C,TG and TC were 0.640,0.827,0.875,0.996 and 0.928,respectively（P=0.000）. Traditional biochemical determination methods for determining sd-LDL-C level and NMR for determining LDL5-C,LDL4-C+LDL5-C+LDL6-C,LDL5-C+LDL6-C and LDL6-C were positively correlated（r values were 0.326,0.341,0.352 and 0.800,respectively,P<0.05）. NMR for determining sd-LDL-C,LDL-C,HDL-C and TC levels had statistical significance compared with traditional biochemical determination methods（P=0.000）. According to the LDL-C and LDL-P determination results of NMR,the percentiles of 54 samples were calculated. Patients with LDL-C percentile≥LDL-P percentile had higher LDL6-C,sd-LDL-C and LDL6-P levels than those with LDL-C percentile<LDL-P percentile（P<0.05）.Conclusions There is a correlation between NMR and traditional biochemical determination methods. There is a certain difference in sd-LDL-C levels in patients with inconsistent LDL-P and LDL-C.

Establishment of influenza A H1N1 virus RNA determination method and its clinical application

TANG Jun, CAO Hongmei

2021, 36(7):  738-742.  DOI: 10.3969/j.issn.1673-8640.2021.07.013

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Objective To establish a real-time fluorescence quantitative reverse transcription-polymerase chain reaction（RT-PCR） for influenza A H1N1 virus RNA,and to investigate its clinical application value. Methods Specific primers and TaqMan probes were designed according to nucleotide sequences of HA and NA genes of influenza A H1N1 virus and nucleotide sequences in GenBank database. The system and conditions in the experimental reaction were improved,and the RT-PCR was established for influenza A H1N1 virus RNA. Its sensitivity,specificity and repeatability were evaluated. Totally,21 samples were collected,all of which were nasopharyngeal swab samples. A comparative study was conducted with chicken embryo culture method.Results The results of RT-PCR were different among different viruses. The RNA of influenza A H1N1 virus was positive,and the RNA of H1-H16 influenza viruses,seasonal influenza virus and swine influenza virus was negative. The sensitivity was 103 copies/μL RNA molecule,and the standard curver ² was 0.998. The cycle threshold（Ct） value within-run and between-run coefficients of variation（CV） of the positive RNA standard were <10%,and the Ct value of the negative RNA standard [diethyl pyrocarbonate（DEPC）] was <0,and the repeatability was good. In the 21 samples,only 2 samples were positive,and the remaining samples were all negative. Compared with chicken embryo culture method,the consistency was 100%. Conclusions The established RT-PCR for influenza A H1N1 virus RNA is simple,time consuming,specific and sensitive,which can be widely used in clinical practice.

Prevalence and genetic characteristic of Staphylococcus aureus with agr dysfunction

BAO Taoxiang, SONG Haoyue, YANG Han, HE Chunyan, SHU Wen, CHEN Wenjing, TANG Rong, LIU Qingzhong

2021, 36(7):  743-748.  DOI: 10.3969/j.issn.1673-8640.2021.07.014

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Objective To investigate the prevalence and genetic characteristic of Staphylococcus aureus with accessory gene regulator（agr） dysfunction. Methods A total of 1 238 isolates of Staphylococcus aureus were collected from 5 hospitals in Shanghai and Zhejiang. All the isolates were determined for methicillin resistance by cefoxitin disc,and were screened for agr dysfunction by determining δ-hemolysin production deficiency. The molecular characteristics of isolates with agr dysfunction were analyzed by agr typing,staphylococcal cassette chromosome mec（SCCmec） typing,staphylococcal protein A（spa） typing and multilocus sequence typing（MLST）. Results The 3.6%（45/1 238） of isolates were agr defective,which was more prevalent among methicillin-resistant Staphylococcus aureus（MRSA）[5.0%（28/565）] and methicillin-sensitiveStaphylococcus aureus（MSSA）[2.5%（17/673）]（P=0.023）. The analysis of genotyping displayed that ST5（CC5）-SCCmecⅡ-t002/t311/t2460-agr2 and ST239（CC8）-SCCmecⅢ-t037-agr1 were the predominant clones among agr defective MRSA isolates,and ST188（CC1）-t189-agr1 was the most abundant clone among agr defective MSSA isolates.Conclusions The rate of Staphylococcus aureus with agr dysfunction is low in Shanghai and Zhejiang,and its prevalence might be related to the spread of specific clones.

Application of Unity Real Time software to establish individual quality control rules for blood cell analysis

ZHANG Hongwei, ZHAO Zhengjie, ZHAO Yan, LI Haisheng, LI Xiufang, QIAN Jing, GUO Chong

2021, 36(7):  749-752.  DOI: 10.3969/j.issn.1673-8640.2021.07.015

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Objective To evaluate blood cell analysis projects through using 6 sigma （σ） performance evaluation,to select the individual quality control rules in order to improve the determination quality.Methods By using the function of Unity Real Time software platform 's Westgard Advisor,the information of σ value,σ measurement figure and suggested rules was obtained according to imprecision [coefficient of variation（ CV）],inaccuracy [bias（Bias）] and allowable total error（TEa）. The projects whose σ values did not reach 6 were analyzed,and the main causes of poor method analysis performance were evaluated through quality goal index （QGI）. The project analysis length was identified according to the analysis length of Westgard σ flow chart of Westgard σ rules.Results The performance characteristics and recommended rules of each item of blood cell analysis are not the same. The selection of individual quality control rules for each item was realized by a single rule or multiple rules. To analyze the projects whose σ values did not reach 6,QGI indicated that,except hemoglobin,accuracy improvement was the priority,while the other items indicated precision improvement was the priority. The length of the analysis batch determined by the comprehensive evaluation of system was 200,which was consistent with the daily sample size of system of about 200. Conclusions The Unity Real Time software platform can be applied to perform 6σ performance evaluation for quantitative analysis projects such as blood cell analysis,select individual quality control rules according to performance characteristics,and make continuous improvement according to QGI. According to the flow chart of Westgard σ rules,the appropriate batch length is determined to ensure the quality of patient test results.

Mechanism and application of serum amyloid A in clinical diseases

LIU Jinyan, ZHAO Juntao, XIANG Mingjie

2021, 36(7):  756-760.  DOI: 10.3969/j.issn.1673-8640.2021.07.017

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Serum amyloid A（SAA） is an acute phase-reactive protein that rapidly increases in the acute phase of infection,inflammation and tissue damage,and rapidly declines during the recovery phase of the diseases. There are important links between SAA and infection caused by bacteria or viruses,atherosclerosis,tumors,transplant rejection,rheumatoid,amyloidosis and so on. This review focuses on the structure,function and determination method of the SAA superfamily and the development of mechanisms and applications of SAA and clinical diseases.

Research progress on Chinese and western medicine treatment for Candida glabrata infection

ZHANG Guanyi, YAO Dongting, WANG Jian, HU Xiaobo

2021, 36(7):  761-765.  DOI: 10.3969/j.issn.1673-8640.2021.07.018

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Candida glabrata is the most common non-Candida albicans,and its resistance rate to azoles is increasing year by year,which brings certain difficulty for the clinical treatment of candidiasis. Studies on genes related to resistance of Candida glabrata find that biofilm-related genes and efflux genes are closely related to drug resistance. In order to solve the problem of drug resistance,the drug resistance mechanism of Candida glabrata and the combination of traditional Chinese medicine in the modern clinical treatment of Candida glabrata infection provides a approach of therapy for the clinical treatment of Candida glabrata infection.