新型冠状病毒肺炎(COVID-19)是由新型冠状病毒(SARS-CoV-2)引起的传染性极强,主要经飞沫和接触传播,不排除通过气溶胶和消化道传播的急性传染病.COVID-19有发热、咳嗽、呼吸急促等症状.反转录聚合酶链反应(RT-PCR)检测SARS-CoV-2核酸是目前确诊COVID-19的重要方法,对判断病情严重程度、是否能出院和解除隔离有重要意义.不同类型标本和不同品牌试剂的核酸检测结果不完全一致.血清学检测SARS-CoV-2阳性率比较高,将核酸检测和血清学检测结合,可提高检出率.

目的 分析低血糖假性危急值发生的原因,寻找有效的改进措施,降低假性危急值发生率.方法 分析2014年7月1日至2015年6月30日住院患者低血糖危急值,鉴别假性危急值并分析其发生原因,采用对策性鱼骨图的方法制订不同的纠正改进措施.结果 78例危急值中,假性危急值27例(34.62％).27例假性危急值中,标本溶血5例(18.52％),静脉滴注(生理盐水)过程中采集标本1例(3.70％).标本放置时间过长(≥120 min)21例(77.78％).通过采取缩短标本周转时间(TAT)、使用血糖专用孵化管、培训采集人员等纠正方式后,假性危急值比例下降至14.90％,与纠正前比较差异有统计学意义(P<0.05).结论 低血糖危急值中存在一定比例假性危急值,应引起临床科室和检验科重视.通过缩短TAT等措施可有效降低假性危急值发生率.

目的 原核表达结核分枝杆菌标准菌株PPE36-p27蛋白,并应用于结核特异性细胞免疫检测.方法 以结核分枝杆菌H37RV基因组DNA为模板,采用PCR法扩增PPE36基因,纯化回收后插入至原核表达载体pEASY,将测序正确的重组质粒转入感受态E.coli BL21(DE3),诱导表达PPE36-p27蛋白,经SDS-PAGE鉴定及NI-NTA亲和层析柱纯化.将纯化后的PPE36-p27作为特异性刺激原,建立新型T细胞检测方法ELISPOT-IL-12技术.结果 重组质粒pEASY-PPE36经双酶切及测序鉴定,构建正确.PPE36-p27蛋白的相对分子质量约30 000,纯化后蛋白浓度为14.69 μg/mL.重组蛋白PPE36-p27应用于ELISPOT-IL-12检测方法时,可刺激机体产生细胞因子IL-12,与包被的抗IL-12抗体产生特异性结合,可见明显斑点.结论 PPE36-p27蛋白在原核表达系统成功表达,以其为刺激原建立了ELISPOT-IL-12检测技术,可应用于结核的早期诊断.

目的：分析甘肃地区乙型肝炎病毒（HBV）的基因型分布。方法采用型特异性引物‐聚合酶链式反应（SSP‐PCR）技术对300例HBV DNA阳性患者进行 HBV基因分型。结果在300份血清标本中，279份标本被成功分型，其中基因型B 59份，占19．7％；基因型C 169份，占56．3％；混合基因型B／C 47份，占15．7％；混合基因型C／D 4份，占1．3％；未确定型别21份，占7．0％。不同基因型 HBV感染患者间性别、年龄分布及丙氨酸氨基转移酶（ALT）水平比较，差异均无统计学意义（P＞0．05）；B型HBV感染患者乙型肝炎e抗原（HBeAg）阳性率（40．7％）略高于C型患者（35．5％），差异无统计学意义（P＞0．05）。结论甘肃地区HBV基因型包括B、C、B／C、C／D型，其中C型为优势基因型。

目的:探讨ERα-29位基因多态性与HBV相关原发性肝癌(PHC)易感性的关系.方法:通过聚合酶链反应-限制性片段长度多态性法检测我院100例HBV相关原发性肝癌患者(PHC组)与95例同期健康体检者(对照组)的ERα-29位基因多态性并对两组间各等位基因、基因型频率进行比较.结果:PHC组ERα-29位基因的TT、TC和CC基因型分别有31例(31.00％)、45例(45.00％)和24例(24.00％),T与C等位基因频率分别为53.50％和46.50％,对照组ERα-29位基因的TT、TC和CC型基因型分别有11例(11.58％)、39例(41.05％)和45例(47.37％),T与C等位基因频率分别为32.11％和67.89％,两组间基因型分布、等位基因频率差异均具有统计学意义(P＜0.05);T等位基因发生PHC的风险是C等位基因的2.43倍(OR =2.43,95％CI:1.37～4.32).结论:ERα-29位基因与HBV相关原发性肝癌易感性有关,其中T等位基因可增加发生风险.

目的 制备乙型肝炎病毒(hepatitis B virus,HBV)DNA定量检测室内质控品,并进行初步评价.方法 对HBV各个基因型序列进行多序列比对分析,选择保守序列S区设计引物,PCR扩增HBV S区基因,与pEASY-T1载体连接,构建重组质粒.测序后,将重组质粒稀释成107 copies/ml、104 copies/ml两个水平,分别作为高值质控品(HBV-H)和低值质控品(HBV-L),分装冷冻于-80 ℃,并初定靶值.评价自制质控品的精密度和稳定性,绘制室内质控图.结果 构建的重组质粒测序结果与目的片段一致,自制HBV DNA 2个水平室内质控品的精密度、稳定性均达到实验要求.结论 自制HBV DNA 2个水平的质控品可用于HBV DNA定量检测的室内质控.

Objective To develop a real-time fluorescent quantitative PCR system in identification of helicobacter pylori ( H.pylori) with characteristics of celerity, sensitivity and specificity.Methods SYBR Green was used in detection of o-ral H.pylori in the real-time fluorescent quantitative PCR system.The results from the system were compared with that from clinical routine detection methods, which could be applied for evaluating sensitivity, specificity and repeatability for the system.Results The data collected from 48 samples revealed that both sensitivity and specificity of real-time fluorescent quantitative PCR are 100%;This system could detect a minimum of 102 copies plasmids;both results from within group re-peating experiments and between groups repeating experiments were corresponding with the data from traditional methods. Conclusion Real-time fluorescent quantitative PCR system has a high sensitivity, high specificity and good repeatability in quantitative determination of oral H.pylori, which is also fast and easy to operate.There fore, this approach will be used to screen oral H.pylori.

目的 分析多发性骨髓瘤(multiple myeloma,MM)患者骨髓细胞形态和免疫型特征,为临床准确诊断提供依据.方法 采用病例对照研究.收集2007年4月至2013年6月甘肃省人民医院初诊的MM患者88例(MM组),同时以88例增生性贫血患者为对照(贫血组).用免疫固定电泳测定其单克隆抗体.分别用光学显微镜和四色流式细胞术检测分析其骨髓细胞形态和免疫型,并比较不同百分比的骨髓瘤细胞组间免疫型表达,以评价MM组免疫型特征.结果 88例MM患者经免疫固定电泳检测,IgG型52例、IgA型26例、IgM型4例,Kappa轻链型52例、Lambda轻链型30例,6例阴性,贫血组无单克隆抗体表达.MM组形态学与贫血组(中位数0/200个)相比,前者浆细胞中幼稚细胞偏多,外形、胞浆、胞核均发生改变,且骨髓瘤细胞比例明显增高(中位数25/200个,t=14.177,P＜0.05).流式细胞术免疫分型结果:MM组CD38+ 98.86％(87/88例)、CD138+ 97.73％(86/88例)、CD56+ 76.14％(67/88例)、CD184+ 73.86％(65/88例),部分伴HLA-DR、CD19、CD20抗原表达,阳性率分别为44.32％(39/88例)、18.18％(16/88例)、15.91％(14/88例),MM组与贫血组比较,差异均有统计学意义(P＜0.05);其他抗原分子(CD5、CD33、CD11b)不表达.用胞质染色法检出Kappa轻链阳性52例[59.09％(52/88例)]、Lambda轻链阳性30例[34.09％(30/88例)],与贫血组比较[0.00％(0/88例)],差异均有统计学意义(P=0.000),且与免疫固定电泳的结果相一致(Kappa值=-0.199,P=0.03).此外,不同百分比的骨髓瘤细胞组间免疫型表达CD38、CD138、CD56、CD184、HLA-DR、CD19、CD20、Kappa、Lambda抗体的组间差异均无统计学意义(Fisher检验,P值分别为0.227、0.230、0.778、0.789、0.113、0.194、0.292、0.938、0.809).结论 骨髓细胞形态学是MM诊断的基本手段,结合免疫分型检测相关抗原分子,可协助临床客观准确地诊断MM.

Objective To explore the concentration of mannan-binding lectin(MBL ) of patients with chronic hepatitis B virus (HBV) .Methods Serum MBL concentrations of 250 patients(case group) with HBV and 150 healthy controls(control group) were measured .Results The serum MBL concentration in case group was higher than that in control group(t=7 .097 ,P<0 .01) . The serum MBL concentration in high HBV-DNA loading group was higher than that in control group(t=7 .179 ,P<0 .01) .The serum MBL concentration in low HBV-DNA loading group was higher than that in control group(t=4 .404 ,P<0 .01) .Conclusion Detection of serum MBL in patients with HBV will be clinically useful for understanding state of an illness and observing the cur-ative effect .

Objective:To analyse mannan-binding lectin (MBL) gene polymorphisms of Han nationality in Gansu province in order to further research about relationship of MBL gene mutation and diseases . Methods: Three polymorphic loci in MBL of 250 healthy Chinese Han individuals from Gansu province were analyzed by PCR-SSP in which the genotype and allele frequency of MBL in different Han population from China were compared . Results: We only found the A and B allele . The genotype frequencies of MBL gene were 74.80%(GGC/GGC),22.00%(GGC/GAC) and 3.20%(GAC/GAC).The allele frequencies of MBL gene were 0.858 (GGC),0.142(GAC). Conclusion: The distribution of polymorphism in MBL gene is consistent with the Hardy-Weinberg equilibrium which is significantly different among the Han population of others areas .

类风湿关节炎（rheumatoid arthritis，RA）是一种严重的病因不清的全身性自身免疫性疾病，临床以滑膜、关节的炎症为特点，进行性侵蚀关节，最终导致残疾和丧失劳动力[1]。已证实RA的关节损伤和慢性炎症有关联，且与炎性指标红细胞沉降率（erythrocyte sedimentation rate，ESR）和C反应蛋白（C-reactive protein，CRP）有关。但部分研究发现RA在炎症活动度下降的情况下，也可能出现关节损伤，无明显炎症症状的患者病情也可能进展到关节侵蚀阶段[2]。因此，早期、可靠地诊断非炎症体征的RA关节损伤非常重要。抗环瓜氨酸多肽（cyclic critrullinted polypeptide， CCP）抗体在诊断RA中有较好的特异度（81%~100%）和一定的敏感度（40%~80%），以及较好的阳性预测值和阴性预测值[3-4]，且可预测早期RA关节损伤[5]。但既往主要为RA的抗-CCP抗体的定性研究，定量研究甚少。我们选择关节侵蚀和非关节侵蚀的RA患者，通过定量测定其抗-CCP抗体水平，并比较分析其和RF、ESR、CRP与关节损伤的关系，探讨其诊断RA关节炎侵蚀的临床意义。

幽门螺杆菌（Hp）是慢性胃炎、消化性溃疡、胃黏膜相关淋巴组织淋巴瘤和胃癌的主要致病因素，1994年被世界卫生组织列为Ⅰ类致癌因子。多年以来，人们普遍认为胃是 Hp 主要的生长环境，而大量研究证明人口腔内牙菌斑、唾液、舌苔中也可检测出 Hp，并可能是胃内 Hp 复发再感染的来源。这对研究 Hp 的传播途径和治疗方法有着重要的意义。

OBJECTIVETo determine the prevalence of saliva Helicobacter pylori in Lanzhou and investigate Helicobacter pylori-related diseases.METHODSHelicobacter pylori was detected through bacterial culture, Gram stain microscopy, and urease test from saliva samples collected from 941 residents of Lanzhou. The infection rate and growth of Helicobacter pylori among the residents were analyzed in terms of different oral health conditions, oral disease, gender, urban and rural status, and age.RESULTSThe rate of Helicobacter pylori-positive saliva in Lanzhou was 42.72%. The status of Helicobacter pylori infection showed significant difference among subjects with different oral hygiene and oral diseases. The rate of Helicobacter pylori-positive saliva among females was 47.89%, which was greater compared with the rate among males (38.45%, P = 0.004, chi2 = 8.492). The rate of Helicobacter pylori-positive saliva in the town was 33.99%, which was less than the rate for the villages (50.93%, P = 0.000, chi2 = 27.551). The rate of Helicobacter pylori-positive saliva among residents aged 10 to 59 showed a flat trend with no significant differences. However, the rate of Helicobacter pylori-positive saliva among residents over 60 years old showed a significant increase. No significant difference was found in the growth of saliva Helicobacter pylori (P = 0.086).CONCLUSIONThe rate of Helicobacter pylori-positive saliva is related to the subjects' oral hygiene, oral disease, gender, age, and living conditions.

目的 按照《血细胞分析仪的校准指南》使用新鲜血作为校准物的校准方法,探讨使用可溯源的定值抗凝新鲜人血对基层医院实验室间血细胞分析仪不同检测系统进行统一比对的应用价值.方法 选用Sysmex XT2000i血细胞分析仪作为参比仪,对抗凝新鲜人血液标本进行定值后分装发放到庆阳市所辖区县16家二级以上医院,对18台血细胞分析仪进行统一比对.结果 三次比对后,项次合格率分别由31.1％,56.7％,68.4％上升到65.2％,87.4％,94.4％,差异有统计学意义(P＜0.01).结论 使用可溯源的定值抗凝新鲜人血对基层医院血细胞分析仪不同的检测系统进行实验室间统一比对,是保证血细胞分析仪检测结果可比性和准确性的有效途径之一,且经济有效.

>分化群抗原64(cluster of differentiation antigen,CD64)具有识别免疫球蛋白、高亲和IgG单体、介导体液免疫和细胞免疫及早期诊断感染性疾病IgG Fc片段受体1(FcγRⅠ)的功能,可通过抗体依赖性细胞毒和细胞吞噬及免疫复合物清除作用清除病原微生物。中性粒细胞CD64是监测感染、抗生素疗效的指标。感染是类风湿关节炎(rheumatoid

Objective To investigate the association between gene polymorphisms in mannan-binding lectin(MBL)and HCV infection.Methods 150 Han ethnic patients with chronic hepatitis C disease and 75healthy controls were studied.The polymorphisms were perfotmed by PCR-SSP.Results The difference in MBL gene genetype and allele distributions between patients with chronic HCV infection and controls was not significant.MBL gene was significantly different at position 54genotypes and allele distributions between ALT>80U/L than in those with ALT<80U/L groups(P<0.05).MBL gene was significantly different at position 54genotypes and allele distributions between medium high and low HCV RNA loading groups.Conclusion The polymorphisms in MBL gene at position 54was not associated with HCV infection.The genetic polymorphisms of MBL may be related to inflammatory reaction in the liver of the patients with chronic HCV infection.

目的 研究内皮祖细胞(EPCs)、C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、纤维蛋白原(FIB)及白细胞(WBC)五项指标在急性胰腺炎(AP)患者及健康志愿者外周血中的分布水平.分析五项指标在重症型急性胰腺炎(SAP)、轻症型急性胰腺炎(MAP)患者外周血中的水平是否存在某种相关性.方法 采集60例AP患者(分为MAP组和SAP组)及20例健康志愿者(对照组)的外周血.采用流式细胞术、ELISA、免疫比浊法等实验方法,检测了各组外周血中EPCs、CRP、TNF-α、FIB及WBC五项指标的水平,采用Spearman′s 相关分析对这五项指标间的相互关系作出评估.结果 TNF-α、WBC、FIB及CRP水平在对照组、MAP及SAP组中,依次升高,具有统计学差异(P＜0.05).SAP组的EPCs水平明显高于MAP组(P＜0.01),而在MAP组与对照组间EPCs的水平没有统计学差异(P＞0.05);MAP组及SAP组外周血中,EPCs、TNF-α、WBC、FIB及CRP水平之间呈正相关.结论 在五项指标中,EPCs和CRP在AP早期预测SAP最具有价值.EPCs可能成为一种新的、具有潜力的生物学指标预测SAP.

Objective To compare the difference of the concentration of immunoglobulins(Ig)and complements between active rheumatoid arthritis(RA)and RA combined with infection and explore the relation between Ig/complement and active periods. Methods The serum concentration of IgG,IgA,IgM and complements C3,C4 were detected in 80 patients with rheumatoid arthritis,which were divided into groups of inactive RA,active RA and RA combined with infection.70 age-matched individuals were included as controls.The correlation of Ig/complement in active RA and RA combined with infection were compared.Results The concentration of serum IgG and IgA levels in active rheumatoid arthritis were significantly higher than that in healthy control and inactive RA.The concentration of IgM was significantly higher than that in control but not than inactive RA.However,there was no significant difference between the active RA and RA combined with infection either in Ig or complements.It was highly correlative between IgG,IgA and RF,CRP or ESR.Conclusion Serum IgG,IgA levels can be used as predictive markers for RA activity;IgG,IgA,IgM cannot be used as an indicator of RA active period and combined infection;complements cannot be used as indicator for RA combined infection,RA active period or RA inactive period.

Objective To investigate hepatitis C virus(HCV) genotypes in patients with viral hepatitis type C in certain area.Methods ELISA and immunoblotting assay were performed for the screening and confirmation of anti-HCV in serum samples,and the virus load in confirmed samples were detected by real-time quantitative PCR.Samples with virus load more than 103 copy/mL were analyzed for HCV genotypes by using multiplex PCR.Results Among 125 cases of samples,positive with anti-HCV,the detection rates of HCV genotype 1b,2a,3a,1c and 2c were 58.4%,21.6%,3.2%,4.0% and 8.0%.Conclusion The most common HCV genotype in patients with HCV infection was 1b,followed by 2a and 3a,and genotype 1c and 2c,rarely detected in other districts,were also could be detected,indicating that the geno types of hepatitis C in this area might be diverse.