Recently, the prevalence of macrolide-resistant Moraxella catarrhalis has been reported, especially among Chinese children. The fitness cost of resistance is reported to render the resistant bacteria less virulent. To investigate the correlation between macrolide susceptibility of M. catarrhalis and pathogenicity, the whole genome of 70 M. catarrhalis isolates belonging to four clonal complexes with different macrolide susceptibilities was sequenced. The gene products were annotated with the Gene Ontology terms. Based on 46 extracted essential virulence genes, 19 representative isolates were selected to infect type II alveolar cells (A549 cells). The ability of these isolates to adhere and invade human epithelial cells and to produce cytokines was comparatively analysed. Furthermore, mice were infected with a pair of M. catarrhalis isolates with different pathogenic behaviours and macrolide susceptibilities to examine pulmonary clearance, histological findings, and the production of cytokines. The percentages of annotations for binding, metabolic process, cellular process, and cell were non-significantly different between the macrolide-resistant and macrolide-susceptible groups. The presence of uspA2, uspA2H, pilO, lbpB, lex1, modM, mboIA, and mboIB significantly differed among the four clonal complexes and macrolide susceptibility groups. Furthermore, compared with those in macrolide-susceptible isolates, the adhesion ability was stronger (P = 0.0019) and the invasion ability was weaker (P < 0.0001) in the macrolide-resistant isolates. Mouse experiments revealed that pulmonary macrophages elicit immune responses against M. catarrhalis infection by significantly upregulating the Csf2, Il4, Il13, Il1b, Il6, Tnf, and Il18. Therefore, M. catarrhalis populations exhibited diverse pathogenicity in vitro and in vivo.

目的 了解2010—2019年北京协和医院肺炎支原体感染的实验室和临床特征,以及儿科患者使用不同药物进行治疗的转归.方法 回顾性收集2010年1月至2019年12月北京协和医院肺炎支原体感染患者的实验室和临床资料,分析不同季节、年龄、来源科室和疾病分布患者的肺炎支原体抗体阳性率,比较儿科住院患者大环内酯类、头孢菌素类、皮质类固醇类药和对症治疗等不同治疗方式的疗效和转归.结果 12985例患者中,15.0％(1950/12985)肺炎支原体抗体阳性;女性患者肺炎支原体抗体阳性率明显高于男性,差异有统计学意义(P<0.05).肺炎支原体感染高峰的季节为秋季.不同年龄段中,>5～10岁患者肺炎支原体抗体阳性率最高(P<0.05).与其他疾病患者相比,社区获得性肺炎患者肺炎支原体抗体阳性率最高(P<0.05).254例儿科肺炎支原体感染住院患者中,有176例、12例、7例和59例患者分别接受了大环内酯类、头孢菌素类、皮质类固醇类药和对症治疗.大环内酯类治疗组只有10例治疗效果不佳,其余均痊愈或改善,其他治疗方案均好转.结论 肺炎支原体感染在季节、年龄、患者科室和疾病分布等方面具有一定的特性,肺炎支原体感染实验室和临床大数据的回顾性分析具有重要意义,可为临床诊疗提供依据.

The mortality rate of ovarian cancer is increasing and the role of hypoxia inducible factor-1α (HIF-1α) in tumor progression has been confirmed. von Hippel-Lindau tumor suppressor protein (pVHL) binds HIF-1α and mediates proteasome degradation of HIF-1α. Besides, histone deacetylase inhibitor (HDACi) mitigates tumor growth via targeting HIF-1α, whereas underlying mechanism still requires investigation. In this research, we exposed ovarian cancer cell lines OV-90 and SKOV-3 to escalating concentrations of HDACi LBH589. As a result, cell viability was significantly suppressed and expression of HIF-1α was remarkably reduced along with decreased levels of signal molecules, including phosphoinositide 3-kinase (PI3K) and glycogen synthase kinase 3β (GSK3β) (P = 0.000). Interestingly, pVHL was expressed in a notably declining tendency (P = 0.000). Chaperone heat shock protein-70 (HSP70) was expressed in an ascending manner, whereas expression of chaperonin TCP-1α was reduced clearly (P = 0.000). Besides, co-inhibition of pVHL plus HDAC did not contribute to a remarkable difference in HIF-1α expression as compared with single HDAC inhibition. Furthermore, both cell lines were transfected with plasmids of VHL plus VHL binding protein-1 (VBP-1). Consequently, the expression of HIF-1α as well as lactate dehydrogenase-A (LDHA) was remarkably decreased (P = 0.000). These findings indicate HDACi may repress expression of HIF-1α via inhibiting PI3K and GSK3β and promote degradation of HIF-1α via HSP70, independent of pVHL. Additionally, a sophisticated network of HDAC and chaperones may involve in pVHL quality control.

目的:评估实时荧光聚合酶链反应(RT-PCR)方法检测腺病毒(ADV)DNA的性能.方法:采集16名健康者咽拭子标本(16例),选取其中10例阴性标本.阳性标本取自1例ADV4型培养液.采用RT-PCR法检测ADV DNA,并依次对检测试剂的准确性、重复性、检测下限、抗干扰能力和特异性等性能参数进行验证及评价.结果:RT-PCR试剂检测ADV DNA的准确性和重复性均为100％,检测下限为5×102 copies/ml.加入100 g/L血红蛋白,药物阿莫西林(5μg/ml)、对乙酰氨基酚(0.15μg/ml)和布地奈德混悬液(0.5 mg/ml)干扰物质后,ADV DNA的Ct均值小于未加入干扰物质的ADV DNA Ct均值的95％的置信区间(95％CI)上限25.51,加入干扰物质未对ADV DNA检测结果造成显著影响.该试剂检测甲型H1N1流感病毒、巨细胞病毒、肺炎链球菌、金黄色葡萄球菌、绿脓杆菌、流感嗜血杆菌和鼻病毒等常见的呼吸道病毒和细菌无交叉反应,具有较好的特异性.结论:RT-PCR法检测ADV DNA的准确性高、重复性好且灵敏度高,抗干扰能力和特异性均符合相关标准的要求,是临床检测ADV的首选方法.

Objective: Hepatitis B virus (HBV) has a worldwide distribution and remains a leading public health problem in China. Method: Automated chemiluminescence microparticle immunoassay was used to test all five markers of HBV serology in serum samples among 696,048 patients, pregnant women, and normal subjects in Beijing from 2008 to 2018. Results: The overall prevalence of subjects categorized as previous/ occult HBV infection, inactive HBsAg carrier, active HBV infection, HBsAg, HBV susceptible, and immune via vaccination was 29.4%, 4.8%, 1.4%, 6.4%, 33.9% and 30.3%, respectively; men had a significantly higher prevalence of HBV infection than women. The prevalence of HBsAg was around 0.5% in subjects <= 10 years of age, increased dramatically to 3.7% in subjects between 11 and 20 years of age, reached the highest level of 7.9% in subjects between 41 and 50 years of age, and finally decreased to 2.8% in subjects >= 81 years of age. During the 10 years from 2008 to 2018, the prevalence of HBsAg was stabilized at about 6.0%, and indicators of HBV susceptibility, previous/ occult HBV infection, and immunity via vaccination were not further improved, despite the constant implementation of HBV vaccination since 1992. All four age groups (21 - 30y, 31 - 40y, 41 - 50y and 51 - 60y) of the normal adult population were found to have a significantly lower prevalence of HBsAg and HBV susceptibility but significantly higher prevalence of immunity via vaccination compared with corresponding age groups of the sub-total population. Conclusions: Although high coverage has been established among infants and young children, their vaccination alone could not reduce HBV infection in the adult Chinese population quickly. Adult populations with more vaccinated individuals are found to have fewer individuals with HBsAg. Vaccination in adults or at least in high-risk adults is an urgent need to decrease horizontal HBV transmission in China. (C) 2020 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-NDlicense (http://creativecommons.org/licenses/by-nc-nd/4.0/).

目的:对国家食品药品监督管理总局(CFDA)批准的4种国产甲型流感病毒核酸检测试剂性能进行评估和临床确认试验.方法:使用美国病理学家协会(CAP)提供的标准毒株(2份)及北京协和医院就诊患者的300份呼吸道剩余样本,对其中2018年1月间84份患者的鼻咽拭子剩余样本进行临床确认试验.参考美国临床和实验室标准协会文件指南,使用CAP提供的A/H1N1和A/H3N2标准毒株制备样本盘,分别从重复性、重现性、检测下限、抗干扰能力、特异性、稳定性和模拟临床评估等方面的性能对试剂A、B、C和D进行评估;结果:试剂C的稳定性较好但其聚合酶链反应(PCR)酶对血液造成的干扰影响较大;试剂D的重复性、重现性、检测下限、抗干扰能力以及稳定性均未达到本实验室的临床应用需求.试剂D与试剂A、试剂B和试剂C的3种试剂评估结果比较,试剂D的重复性、重现性和抗干扰能力结果存在明显差异,具有统计学意义(F=6.53,F=25.44,F=14.15;P<0.05).试剂A的检测下限可以达到102 copies/ml,在4种试剂中具有较明显优势.临床确认实验中试剂A和试剂B检测结果的阳性率分别为38.1％(32/84)和23.8％(20/84),两者比较差异具有统计学意义(Z=-3.464,P<0.05).结论:通过对重复性、重现性、检测下限、抗干扰能力、特异性、稳定性及模拟临床评估等方面的性能评估和临床确认试验,试剂A漏检率更低,临床检测准确率更高.

目的 对3种不同厂家的商品化沙眼衣原体即时检测(POCT)试剂盒进行评估,为中国医学科学院北京协和医院检验科实验室及其他医疗机构实验室提供参考.方法 采用沙眼衣原体菌株(血清型D和E)冻存毒株,化冻后经细胞培养法滴定确定浓度,使用生理盐水进行系列稀释,对试剂盒A(衣原体抗原检测试剂盒)、试剂盒B(沙眼衣原体抗原检测试剂盒)、试剂盒C(沙眼衣原体抗原检测试剂盒)进行最低检出限和重复性实验.采用人胚肺成纤维细胞、非洲绿猴肾细胞系、大肠埃希菌标准菌株菌悬液进行特异度实验.结果 对血清型D的最低检出限,试剂盒B和C浓度均达1.86×107 IFU/mL,试剂盒B在1.86×106 IFU/mL观察到隐约可见条带;试剂盒A仅能检测到1.86×109 IFU/mL.对血清型E检测灵敏度,试剂盒B和C均达2.80×107IFU/mL,但均为弱阳性;试剂盒A仅能检测到2.80×109 IFU/mL.3种试剂盒均有较好的重复性,但均存在阳性条带显示颜色强弱不完全一致的情况.3种试剂盒均能够抗人胚肺成纤维细胞MRC-5 (2.9×106/mL),非洲绿猴肾细胞系BGMK(5.0×106/mL)和标准菌株ATCC25922大肠埃希菌(1.5×108 CFU/mL)的干扰.结论 试剂盒B和C最低检出限明显优于试剂盒A,但试剂盒B较试剂盒C略好.在特异度和重复性方面3种试剂盒均较好.

目的 分析xTAG脑脊液病毒多重检测试剂盒(cerebral spinal fluid viral panels,xTAG CSFVP)对成人疑似病毒性脑炎、脑膜炎病原体检测结果.方法 收集2014年10月～2016年9月就诊于北京协和医院的115例疑似病毒性脑炎或脑膜炎的成人患者脑脊液标本115例,使用xTAG CSFVP进行检测并对结果进行分析.结果 115例脑脊液标本中,有28例检测出病毒,阳性率为24.3％,其中单纯疱疹病毒1型的检出率最高(7.8％),水痘-带状疱疹病毒和EB病毒检出率次之(均为6.1％).共检出病毒30例,其中单一感染占92.86％(26/28),混合感染占7.14％(2/28).xTAG CSFVP阳性和阴性患者的临床症状和脑脊液生化指标差异无统计学意义(均P>0.05).结论 成人病毒性脑炎脑膜炎患者xTAG脑脊液病毒检出率最多的病原体为单纯疱疹病毒1型,水痘-带状病毒和EB病毒,检出2例混合感染.

BackgroundPrimary cytomegalovirus (CMV) infection is prevalent worldwide and usually results in latency in immunocompetent populations. Reactivation of latent CMV can cause life-threatening complications in immunocompromised hosts.MethodsWe used the CMV Brite assay to test CMV antigenemia (pp65) in whole blood samples from 22,192 patients with or without autoimmune diseases in Beijing during 2008-2018.ResultsThe overall prevalence of CMV antigenemia was 19.5% (9.7%, males; 26.0%, females). The prevalence of CMV antigenemia was 35.1%, 58.6% and 11.4% in whole patients with autoimmune diseases, in patients with systemic lupus erythematosus (SLE) and in patients with non-SLE autoimmune diseases, respectively. All patients with non-autoimmune diseases, patients with HIV/AIDS or transplantation were found to have 5.0%, 27% or 14.8%, respectively. Patients≤20 years with SLE had a significantly higher prevalence of CMV antigenemia than did all SLE patients, on average. Patients>51 years with non-SLE autoimmune diseases had a significantly higher prevalence than did all patients with non-SLE autoimmune diseases, on average. The prevalence of CMV antigenemia in patients admitted to intensive-care units (ICUs) were 9.2%, which was significantly higher than that among all patients with non-autoimmune diseases. Patients with SLE had 23.8% of negative conversion of CMV antigenemia, significantly lower than the percentage of patients with non-SLE autoimmune (64.3%) and non-autoimmune (61.0%) diseases. The mean number of days to negative conversion of CMV antigenemia in patients with SLE was 35.3±35.8 days, which was significantly longer than that in patients with non-SLE autoimmune diseases (15.4±11.9 days) and non-autoimmune diseases (13.6±7.7 days).ConclusionsCMV antigenemia is found more likely in women than in men, more prevalently in patients with SLE than those with HIV/AIDS or transplant recipients, more frequently in patients admitted to ICUs. Patients with SLE had prolonged CMV antigenemia. The role of CMV appears important in SLE.

Chimeric antigen receptor (CAR)-T cells reprogram the T cells to target tumor cells and have undergone several generations of enhancement. Accumulating evidence shows that by incorporating co-stimulatory molecules and single-chain variable fragments (scFvs), CAR-T cells maintain sustainable proliferation capacity along with high sensitivity and enhanced cytokines production. Moreover, CAR-T cell therapy has shown great promise in clinical trials. However, complications and toxicities must be considered and managed. In this review, we aimed to summarize the development of CAR and the criteria for the selection of the tumor associated antigen (TAA) based on the literature. In addition, we review current clinical trials of CAR-T therapy. Finally, we discuss the adverse effects of CAR-T therapy, managements of complications, and expectations from CAR-T therapy.

BACKGROUND:Epstein-Barr virus (EBV) is associated with nasopharyngeal carcinoma (NPC) which is prevalent in South China, and its association with systemic lupus erythematosus (SLE) or other autoimmune diseases has not been studied in the mainland of China. The EBV serological tests have been performed on patients with various diseases or manifestations for years at our institution and their values need to be evaluated.METHODS:For routine medical purposes, anti-EB viral capsid antigen (VCA) IgG, IgA and IgM antibodies, anti-EBV diffuse early antigen (EA-D) IgA antibodies, and anti-EBV nuclear antigen-1(EBNA-1) IgG antibodies were tested with commercial enzyme-linked immunosorbent assay (ELISA) in patients visiting Peking Union Medical College Hospital between 2013 and 2017. The test results were analyzed in this retrospective study.RESULTS:There were a total of 11122 serum samples available to be tested in the study. As indicators of past EBV infection, the prevalence of VCA-IgG/EBNA1-IgG were 66.6%/58.5%, 84.3%/78.8%, 92.9%/87.0% and 98.5%/95.4% in patients aged under 5 years, 6-10 years, 11-20 years and 21-30 years old, respectively, and these values maintained at this highest rate as age increased further. The prevalence of VCA-IgM, as a parameter of acute EBV infection, was 14.6%, 10.2%, 10.4%, 6.3% and 3.1% in patients aged under 5 years, 6-10 years,11-20 years, 21-30 years, 31-40 years old, respectively, and decreased to 2%~3% in older patients. Patients with elevated serum liver enzymes were more likely to have a higher prevalence of EA/D IgA antibody (P < 0.01) and young patients (≤30 years) with lymphadenopathy were more likely to have higher prevalence of VCA-IgM antibody (P < 0.01). The prevalence of VCA-IgA and EAD-IgA were 87.0% and 59.2% in NPC patients, respectively, and both were significantly higher (P < 0.001) than that in non-NPC patients. The prevalence of VCA-IgA was 45.4% and 25.6% in SLE patients and patients with other autoimmune diseases, respectively, which were significantly (P < 0.001) and mildly (P = 0.039) higher than their controls. In pediatric SLE patients between 6 and10 years old, the prevalence of VCA-IgG, VCA-IgA and EBNA1-IgG was 100%, 59.5% and 100%, respectively, all being significantly higher than the age (6-10y) related controls (P< 0.01). In the 705 cerebral spinal fluid (CSF) specimens, VCA-IgG, VCA-IgM, VCA-IgA and EAD-IgA were found to be positive in 12.1%, 0.15%, 0.25% and 0.25%, respectively. There were 157 paired specimens (CSF and serum were collected simultaneously) and VCA-IgG was identified as positive in 12.7% of the CSF and 100% of the serum specimens.CONCLUSIONS:Around 98% of Chinese patients were infected with EBV before 30 years of age and the highest rate of acute EBV infection were observed in patients under 5 years old. EBV infection was found to be associated with elevated serum liver enzymes, NPC and SLE. Acute anti-EBV antibody was valued for young patients with lymphadenopathy but limited value for CNS neuropathy.

Malaria is a kind of disease detrimental to human health and plasmodium is a critical pathogen in it. The immunity against foreign antigens including plasmodium could be divided into two categories, namely, adaptive immunity and innate im-munity. Innate immunity induces non-specific immune response, and is composed of monocyte,macrophage,γδT cell,DC,NK, and cytokines etc. Innate immunity cooperates with adaptive im-munity efficiently to protect against malaria. Meanwhile,autoph-agy is not only the cellular degrading process, but also gets in-volved in regulating immune system and defending against plas-modium infection. Therefore, elucidation of corresponding mechanism could provide proof for efficiently controlling and cu-ring malaria,developing related medicine and vaccine,and clin-ical treatment as well. This article reviews the constitution of in-nate immunity in malaria,related regulation mechanism and rel-evant therapeutic targets for it.

目的 制备抗弓形虫特异性单克隆抗体并初步应用.方法 弓形虫RH株体外细胞培养增殖、纯化后灭活并超声波破碎免疫BALB/c小鼠,小鼠脾细胞与骨髓瘤细胞NS-1进行融合.酶免法筛查阳性杂交瘤并局限性稀释法进行克隆.免疫荧光、组化及印迹对筛选后的克隆进行鉴定,最终获得的杂交瘤制备腹水并使用Protein G进行纯化.结果 2只小杂交瘤融合后初步筛查出约80株阳性克隆.剔除与其它抗原有交叉反应、抗体效价低的克隆,最终获得抗弓形虫单抗杂交瘤1株,即14H4-1.免疫荧光和免疫组化显示该株单抗与弓形虫RH具有良好的反应性,免疫印迹试验显示与弓形虫28kDa抗原条带结合.14H4-1单抗免疫球蛋白亚型为IgG1,Protein G纯化腹水后的单抗效价≥1∶12800.14H4-1单抗已应用排除2例弓形虫感染疑似诊断.结论 制备的抗弓形虫单克隆抗体性能良好,可应用于弓形虫感染病原学检测.

Background Chlamydia pneumoniae (C. pneumoniae) is an obligate intracellular bacterium and a human pathogen that causes respiratory infectious diseases. More than 50% of the adult population worldwide was once infected with C. pneumoniae, but investigations into this topic are insufficient in mainland China. Methods Anti-C. pneumoniae IgG and IgM antibodies were detected using micro-immunofluorescence test in serum samples of patients visiting Peking Union Medical College Hospital between 2008 and 2017 for routine medical purposes, and the aim of this retrospective study was to analyze the test results. Results Among 12,050 serum specimens tested for anti-C. pneumoniae IgG and IgM antibodies, the overall prevalence of anti-C. pneumoniae IgG antibodies was 86.6%, 87.2% for men and 86.0% for women. Adult men (>20 years) were found to have a significantly higher prevalence of anti-C. pneumoniae IgG than women (χ2 = 30.32, P = 0.000). 3 to 5 years old patients were observed to have the lowest prevalence of anti-C. pneumoniae IgG, 42.8%, then increased with age, reaching the highest level of 98.6% in patients over 70 years of age. In the 10,434 specimens with C. pneumoniae IgG antibodies, the total geometric mean titer (GMT) for C. pneumoniae IgG was 45.71. Although GMTs were found to be significantly higher among all men than among all women (t = 5.916, P = 0.000), sex difference actually began in patients over 40 years of age and increased in the elderly. In the total 12,050 specimens, 1.2% had anti-C. pneumoniae IgM, 3.3% had anti-C. pneumoniae IgG with titers equal to or greater than 1:512; 0.39% had ≥4-fold increasing titers of antibodies in acute and convalescent phase paired samples, and 4.4% were finally confirmed to have acute antibodies against C. pneumoniae. 6 to 10 years old patients were found to have the highest rate of both IgM antibodies (3.9%) and acute antibodies (6.2%) against C. pneumoniae. Acute antibodies against C. pneumoniae were found to be more frequent in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD, 14.0%, χ2 = 20.43, P = 0.000), patients with pneumonia (7.8%, χ2 = 51.87, P = 0.000) and patients with acute respiratory tract infection (12.3%, χ2 = 60.91, P = 0.000) than among all patients (4.4%). Both anti-C. pneumoniae IgG and IgM antibodies should be tested for acute antibodies against C. pneumoniae as testing for either alone will underestimate by a maximum of two-thirds the incidence of acute antibodies against C. pneumoniae. Conclusions More than 86% of Chinese patients on an average were once infected with C. pneumoniae. Adult men had both a higher prevalence and higher levels of antibodies than women. 6 to10 year old patients were found to have the most frequent acute infection of C. pneumoniae. C. pneumoniae is associated with AECOPD, pneumonia and acute respiratory tract infection. Anti-C. pneumoniae IgG and IgM should be tested simultaneously to avoid underestimation of acute antibodies against C. pneumoniae.

Cytomegalovirus (CMV) infection is associated with glucocorticoid resistance in ulcerative colitis (UC) and may exacerbate the disease course. However, the underlying pathogenicity remains unclear. The aim of this study was to explore possible underlying mechanisms during CMV latency and lytic infection in the human mononuclear cell line THP‐1. Latent and activated CMV infection cell models were established. We performed real‐time PCR and western blotting to examine changes in glucocorticoid receptors (GRs) during CMV latency and activation. Pro‐inflammatory and anti‐inflammatory cytokines were detected by ELISA. After UV‐inactivated CMV infection, GRs and cytokines were also examined. The expression of GRs was elevated in the reactivation group. An increased ratio of GR β/α and phosphorylation of GRα in the CMV reactivation group may explain refractory response to steroids. During CMV lytic infection, pro‐inflammatory cytokines IL‐6 and TNF‐α increased remarkably and anti‐inflammatory cytokine IL‐5 decreased, which may exacerbate UC. GR and cytokines were unchanged in the UV‐inactivated CMV infection group. Changes in the number and function of GRs may account for glucocorticoid resistance in CMV reactivation. The imbalance of pro‐ and anti‐inflammatory cytokines may be related to severe inflammation.

Objective: The objective was to compare the performance of four commercial rapid influenza virus antigen detection test kits, namely QuickNavi-Flu (Denka Seiken, Tokyo, Japan), Alere BinaxNOW Influenza A&B assay (Alere Scarborough, Scarborough Inc., Scarborough, ME, USA), KaiBiLi Genesis Influenza A&B Antigen Test (Genesis, Hangzhou, China) and Actim Influenza A&B (Medix Biochemica, Kauniainen, Finland). Methods: CAP proficiency testing specimens and ultrasonic lysates of normal human lung fibroblast cell (MRC-5 at 2.9 × 106/ mL) were taken as specimens. The supernatants of influenza A and influenza B were used as stock solutions, and, using saline, were diluted to 1:10 and then serially diluted to 1:100, 1:200, 1:400, 1:800, 1:1600, and 1:3200. The dilutions were tested respectively to evaluate the sensitivity of these kits. Respiratory syncytial virus, adenovirus, Para influenza virus type-2 and MRC-5 cell were taken as samples to evaluate the specificity of these kits. Influenza A (H1N1) at a dilution of 1:200, influenza A (H3N2) at a dilution of 1:100 and influenza B at a dilution at 1:100 were used to detect the reproducibility. Each sample was tested nine times in total. Results: In this experiment, the four commercial kits showed the same detection limit for influenza A (H3N2) where all the kits could not give positive result for dilutions lower than 1:100. On the contrary, different detection limits were obtained for influenza A (H1N1) and influenza B. For influenza A (H1N1), positive results could be obtained using a dilution at 1:800 by Quick Navi Flu yet positive results could only be obtained at the dilution of 1:200 using the other three kits. All the results showed that timely detection gave better sensitivity than delayed detection except detection for Influenza A (H1N1) by the Alere BinaxNOW kit where dried specimens gave slightly higher sensitivity. The Actim kit showed positive result for Influenza B when detecting the ultrasonic lysates of MRC-5 cell, illustrating that false positive result may be obtained when detecting specimens which contain components related to MRC-5 cell using Actim kit. There was no non-specific cross-reactivity for the other three kits. In general, all these kits provided good reproducibility except that results of of Influenza A (H1N1) detection using the Alere BinaxNOW kit were inconsistent. Conclusions: Various performances were found for the detection of influenza A and B viruses using four rapid immunochromatography diagnostic kits most commonly used in China.

Objective To prepare monoclonal antibody against early and late antigens of cytomegalovirus (CMV) and to use the antibody for the identification of isolates from proficiency testing and clinical samples.Methods BALB/C mice were immunized with soluble and formaldehyde to inactivate CMV antigens from the CMV-infected MRC-5 cells at 24,48 and 72 hours after inoculation.Mouse myeloma NS-1 cells were fused with spleen cells of the immunized mice.Positive hybridomas were screened with EIA and were cloned with limited dilution method.The immunofluorescent and immunoblotting tests were applied to confirm the positive clones after screening.The ascites was prepared with the identified hybridoma and was purified by Protein G.The purified monoclonal antibody was used to identify CMV isolates from CAP proficiency testing samples and clinical samples.Results A total of 110 positive hybridomas clones were preliminary identified from fused cells of three immunized mice.One hybridoma clone,23B5-1 was finally obtained while other clones were eliminated due to cross-reactive with other antigens,low antibody titers or without reactions with both CMV early and late antigens.It was shown that 23B5-1 reacted with CMV infected MRC-5 cells from 3 hours to 120 hours (5 days) after inoculation.All of immediate-early,early and the late CMV antigens were recognized by the monoclonal antibodies simultaneously.It was also shown that 23B5-1 reacted with five CMV protein bands between 25KD to 50 KD in western blot.The immunoglobulin subclass of antibody was IgG1 and the titer of the purified antibody was equal or more than 1:12 800.The results of staining with the antibody were consistence with predicted results for all six CMV isolates from proficiency testing and ten isolates from clinical samples.Conclusions The performance of our self-prepared monoclonal antibody was good according to the preliminary application.

Objective To retrospectively study the serum IgG and IgM antibodies against toxoplasma, rubella virus, cytomegalovirus and herpes simplex virus type 1&2 in various populations, and analyze the clinical values.Methods From 2008 to 2015, 2 661 pregnant women, 324 infertile women, 2 492 women with abnormal pregnancy history, 623 women with recent abnormal pregnancy, 261 infants with intrauterine growth retardation and other diseases, 170 women for preconceptual examination, and 702 women for physical examination in Beijing were included .Commercial EIA kits were used to detect serum IgG and IgM antibodies to toxoplasma, rubella virus, cytomegalovirus and herpes simplex virus type 1&2. Positive reactions of IgM antibodies to any pathogens were re-tested with another kind of commercial EIA kit. PEMS3.1 software was used for statistical analysis.Results The prevalence of serum IgG or IgM antibodies against toxoplasma, rubella virus, cytomegalovirus and herpes simplex virus type 1& 2 were found within 0.7%-1.6%(0-1.2%) , 85.3%-92.0% ( 0.4%-2.7%) , 89.1%-94.9% ( 0.7%-1.7%) , 74.8%-86.0% ( 0 -0.7%) , 8.1% -17.4% ( 0 -4.1%) respectively in the studied population groups.The prevalence of TORCH IgG and IgM antibodies were not found to be higher in both populations with past suspicious exposure ( infertile women and women with abnormal pregnancy history ) and recent suspicious exposure ( women with recent abnormal pregnancy and infants with intrauterine growth retardation and other diseases) than that in pregnant women and women for preconceptual and physical examination. Conclusion No associations between TORCH infections and the suspicious exposure were found in the populations above.

Despite the significant advancement in vaccine, influenza continues to be a threaten to public health. Influenza has caused serious morbidities and mortalities both during yearly seasonal epidemics and historic pandemics. Early and accurate laboratory diagnosis to influenza virus is critical for initiation of antiviral therapy with neuraminidase inhibitors and for control of the infection. We discuss currently available approaches of laboratory diagnosis for influenza, in which viral culture, antigen detections, nucleic acid amplification tests and serology are included. Key words: Influenza virus; Viral culture; Antigen detection; Nucleic Acid Amplification Tests; Serology