接种甲型H1N1流感疫苗献血者血清中血凝素IgG抗体的检测

目的研制甲型H1N1流感(甲型流感)血凝素(HA)IgG抗体(抗-HAIgG)检测试剂,并用于对献血人群中甲型流感疫苗(甲流疫苗)接种者的抗-HAIgG检测。方法克隆表达甲型流感HA表位抗原(18-243aa),制备出酶联免疫吸附法(ELISA)抗-HA试剂,并对106份甲型流感流行前的献血者血清(甲型流感前组)、97份接种甲流疫苗后的献血者血清(接种甲流疫苗组)做抗-HAIgG检测。结果甲型流感前组:4例抗-HAIgG为阳性,102例阴性,阳性率为3.77%(4/106);甲流疫苗组:78例抗-HAIgG为阳性,19例阴性,阳性率为80.41%(78/97)。结论ELISA法检测甲型流感抗-HAIgG试剂具有较高的特异性、灵敏度和安全性,其在献血人群中的甲型H1N1病毒感染的流行病学调查、疫苗免疫血浆的筛选方面,具有一定的应用价值。     

应用EIA法检测舟山地区献血者甲型H1N1流感病毒抗体

通常当某一人群中甲型H1N1流感(甲型流感)病毒抗体(抗-H1N1)阳性率>60%时,该人群中不会暴发甲型流感.为了解普通人群对甲型流感病毒的免疫保护情况,更好地应对今年春、秋可能发生的甲型流感疫情,做好其间的无偿献血工作,保障医疗急救用血[1],我们应用EIA法对舟山地区无偿献血者人群的抗-H1N1水平进行了检测,现将结果报道如下.     

甲型H1N1流感病毒抗体血浆治疗危重甲型H1N1流感患者疗效观察

目的探讨甲型H1N1流感病毒抗体血浆用于危重甲流患者的治疗效果。方法采集制备H1N1抗体血浆,选择甲型H1N1流感病毒核酸检测阳性、经省专家组会诊判断为危重的患者,结合患者病情静脉输入200ml抗体血浆,观察其病程变化及治愈情况。结果4名危重甲流患者经抗体血浆联合抗病毒治疗后治愈3例,死亡1例。讨论抗体血浆治疗甲流危重患者有一定的治疗效果,适合于流行病区治疗,但还需进一步模索使用方法。     

浙江省甲型H1N1流感病毒抗体血浆抗体水平检测与分析

2009年3月以来,包括墨西哥、美国和加拿大在内的许多国家发生了人感染甲型H1N1流感(以下简称甲流)病毒疫情.世界卫生组织(WHO)于6月11日将流感大流行的预警级别提高至6级,宣布全球性的流感大流行已经到来.为了有效应对可能发生的甲流大流行,卫生部于2009年9月10日下达了采集甲流病毒抗体血浆工作的通知.2009年10月-12月浙江省采供血机构共采集132份抗体血浆,并随机调查的20名疫苗免疫后献血者,我们对这152份样本进行了抗体效价的检测,现分析如下.     

广州地区无偿献血人群中H1N1和H3N2流感病毒中和抗体的分布及水平

目的 探讨H1N1和H3N2流感病毒中和抗体在广州地区无偿献血人群中的分布及水平,为研究广州地区流感流行病学特征提供参考.方法 于2015年10～11月,采用年龄分层随机抽样法选择广州血液中心无偿献血的498例健康献血者为研究对象.按年龄将其分为18～20岁组(n=150)、21～30岁组(n=210)、31～60岁组(n=138).采用微量病毒中和实验法测定献血者血浆中H1N1和H3N2流感病毒中和抗体滴度,并分别对不同性别及年龄段H1N1和H3N2流感病毒中和抗体阳性率及滴度进行统计学分析.结果 498例健康献血者中,H1N1流感病毒中和抗体阳性率为3.6％ (18/480);H3N2流感病毒中和抗体阳性率为4.6％ (23/498),2种流感病毒中和抗体阳性率比较,差异无统计学意义(x2=0.636,P=0.425).156例男性献血者中,H1N1和H3N2流感病毒中和抗体阳性率分别为3.8％(6/156)和4.5％(7/156);342例女性献血者中H1N1和H3N2流感病毒中和抗体阳性的率分别为3.2％(11/342)和4.4％(15/342).不同性别献血者H1N1和H3N2流感病毒中和抗体阳性率比较,差异均无统计学意义(x2 =0.129、0.003,P＞0.05).18～20岁组、21～30岁组和31～60岁组献血者H1N1流感病毒中和抗体阳性率分别为4.0％、5.2％和0.7％;H3N2流感病毒中和抗体阳性率分别为6.0％、5.2％和2.2％.不同年龄组献血者H1N1和H3N2流感病毒中和抗体阳性率比较,差异均无统计学意义(x2=4.961、2.705,P＞0.05).18～20岁组、21～30岁组和31～60岁献血者H1N1流感病毒中和抗体几何平均滴度(GMT)分别为1∶1.38、1∶1.48和1∶1.05,平均为1∶1.32;H3N2流感病毒中和抗体GMT分别为1∶1.62、1∶1.51和1∶1.19,平均为1∶1.44.3组献血者H1N1和H3N2流感病毒中和抗体滴度比较,差异均无统计学意义(x2 =4.887、2.702,P＞0.05).结论 广州地区无偿献血人群中H1N1和H3N2流感病毒中和抗体水平较低,不同性别和年龄人群对流感病毒普遍易感.     

组织采集甲型H1N1流感抗体血浆的回顾与思考

2009年3月墨西哥和美国暴发了甲型H1N1流感(甲型流感)疫情并迅速在全球蔓延.2009年5月11日我国确诊了内地首例甲型H1N1流感患者[1].世界卫生组织(WHO)于2009年6月11日将甲型H1N1流感大流行警告级别提升为6级.截至2010年1月10日,中国内地已有12万4 764例甲型流感确诊病例(不包括临床诊断病例),其中死亡744例;除海南省外,所有省区都报告了死亡病例.面对突如其来的疫情,为最大限度地减少了重症和死亡病例的发生,卫生部于2009年9月10日下达了<关于开展采集甲型H1N1流感病毒抗体血浆采集工作的通知>[2](简称<通知>).本中心迅速组织浙江全省采供血机构采集甲型H1N1流感病毒抗体(抗-H1N1)血浆,至2010年1月18日,已从无偿献血人群中成功采集132份共31 270 ml的抗-H1N1血浆.现将这一过程做简要的回顾和思考.     

学生接种甲型H1N1流感疫苗免疫原性与持久性观察

目的 通过对学生接种甲型H1N1流感疫苗的免疫原性及持久性观察,为今后甲型H1N1流感疫苗的使用与免疫策略提供参考依据。 方法 选择辖区内某中学初三年级学生为研究对象,以知情自愿为原则,于上臂三角肌处接种一剂国产甲型H1N1流感疫苗,并于接种前、接种后1、3、6个月和1年采集血清,检测不同时间段甲型H1N1流感HI抗体情况。 结果 学生免前甲型H1N1流感HI抗体阳性率为35.68%,免后1个月HI抗体阳转率为75.14%,阳性率为92.43%,GMT(倒数)为177.42,增长9.32倍,免后1年HI抗体阳性率仍在84.21%、GMT(倒数)93.03水平。 结论 国产甲型H1N1流感疫苗具有较好的免疫原性与免疫持久性,免疫后1年不用再次免疫。     

血清超敏C-反应蛋白在甲型H1N1流感患者中的变化及临床意义

目的探讨检测血清超敏C反应蛋白(hs-CRP)在甲型H1N1流感患者中的变化及临床意义。方法检测79例(轻症42例、重症37例)甲型H1N1流感患者的血清hs-CRP水平,分析和同期40例普通流感患者的血清hs-CRP水平比较。结果甲型H1N1流感患者血清hs-CRP阳性率明显高于普通流感组,差异具有统计学意义(χ2=40.83,P<0.05);重症甲流组血清hs-CRP水平明显高于轻症甲流组及普通流感组,差异具有统计学意义(F=22.80,P<0.05)。结论血清hs-CRP的检测可作为甲型H1N1流感的早期诊断、病情监测的辅助指标。     

甲型H1N1流感危重症临床特点及危险因素分析

目的 了解甲型H1N1流感(甲流)危重症患者的临床特点,分析其死亡相关危险因素.方法 采用多中心研究方法,回顾性分析郑州市4家医院2009年10月至2010年2月住院的128例甲流危重症患者的临床数据,比较存活组(110例)与死亡组(18例)患者的临床特点;对年龄>14岁的成人患者,以性别、是否妊娠、基础疾病数量、心率、平均动脉压、pH值、动脉血氧分压(PaO2)、氧合指数(PaO2/FiO2)、是否有呼吸衰竭、是否出现急性呼吸窘迫综合征(ARDS)、单肺或双肺病变、是否使用糖皮质激素作为变量进行二元Logistic回归分析,探讨甲流危重症患者死亡的相关危险因素.结果 128例患者中孕妇病死率显著高于儿童和成人非孕妇患者[37.5%(6/16)比12.5%(4/32)、10.0%(8/80),P=0.044和P=0.0003.死亡组呼吸衰竭、ARDS、有创机械通气、有合并症的比例(分别为83.3%、33.3%、66.7%、77.8%)及心率[(114士25)次/min]均显著高于生存组[37.3%、9.1%、12.7%、51.8%、(101+21)次/min);pH值、PaO2(mm Hg,1 mm Hg=0.133 kPa)、PaO2/FiO2(mm Hg)均显著低于生存组(pH值:7.149±0.404比7.398±0.157,PaO2:58.57±36.85比85.78±36.10,PaO2/FiO2:118.17±105.52比259.25±174.91);使用糖皮质激素的比例多于生存组(77.8%比36.4%,P<0.01),且使用时间(d)明显长于生存组(8.3+8.0比2.64±4.2,P<0.05).Logistic回归分析显不,妊娠[相对比值比(OR)为6.642,P=0.011]及ARDS(OR=10.603,P=0.001)是成人甲流危重症患者的死亡危险因素.结论 在甲流危重症患者中,孕妇的死亡比例高于其他患者;有呼吸衰竭、特别是ARDS患者和需要有创机械通气治疗者、有合并症者的死亡比例显著增加;死亡患者发病初期心率明显增快.糖皮质激素的过度使用可能不利于甲流危重症的控制.妊娠和ARDS是甲流危重症成人患者死亡的高危因素.     

获得高效价甲型H1N1流感病毒抗体原料血浆初探

目的探讨从免疫供血浆者获得高效价甲型H1N1流感病毒抗体血浆,为制备人甲型H1N1流感免疫球蛋白和重症患者临床使用提供依据。方法用批签法核准的市场供应甲型H1N1流感疫苗对1947名供血浆者按规定免疫后第2、4、6周收集血浆并采用血凝抑制试验进行检测。结果经检测供血浆者抗体效价≥10以上达到99.2%,其中≥320—10240达到48.4%。结论供血浆者经一次甲型H1N1流感疫苗免疫后收集高效价甲流抗体血浆用于制备特异性免疫球蛋白或供重症患者临床使用是可行的。     

Procalcitonin levels are lower in intensive care unit patients with H1N1 influenza A virus pneumonia than in those with community-acquired bacterial pneumonia. A pilot study

Purpose

The purpose of the study was to know the kinetics of procalcitonin (PCT), C-reactive protein (CRP), and white blood cell (WBC) in critically ill patients with H1N1 influenza A virus pneumonia and to compare levels of these inflammatory mediators with patients with acute community-acquired bacterial pneumonia.

Materials and Methods

An observational study in a mixed intensive care unit (ICU) at a general university hospital was performed. All consecutive patients admitted to the ICU with a diagnosis of severe acute community-acquired pneumonia from September 2009 to December 2009 were included. Viral (H1N1 influenza A) and bacterial microbiological diagnoses were done in every patient. At admission, demographics, comorbidities, Simplified Acute Physiology Score, Sequential Organ Failure Assessment, Lung Injury Score, and Pao₂/Fio₂ were recorded. At admission and after 24, 48, and 120 hours, WBC, CRP, and PCT levels were obtained. Finally, hospital and ICU length of stay and mortality were recorded.

Results

No differences in CRP or WBC were found between H1N1-positive patients and H1N1-negative patients (patients with acute community-acquired bacterial pneumonia). Procalcitonin levels at admission were lower in H1N1-positive patients (PCT = 0.4 [0.1-6.1] ng/mL) than in the H1N1-negative patients (24.8 [13.1-34.5] ng/mL). Procalcitonin significantly decreased with time but remained lower in the H1N1-positive group at all measurements (P < .05 for all comparisons).

Conclusions

Among patients admitted to the ICU with pneumonia, the PCT level could help identify H1N1 influenza A virus pneumonia and thus enable earlier antiviral therapy.     

山东省部分人群甲型H1N1流感血清抗体水平研究

目的 了解甲型H1N1流感大流行季节山东省甲型H1N1流感抗体水平,分析甲型H1N1流感病毒感染状况。 方法 2010年1月10日、25日、2月20日和3月20日随机选择山东省2家医院门诊就诊人群和血站献血人员分年龄组采集1617人份静脉血,采用血凝抑制试验检测甲型H1N1流感血清抗体水平进行分析。 结果 山东省4次甲型H1N1流感血清抗体阳性率和抗体几何平均滴度(GMT)分别为18.24%(GMT 13.86)、26.95%(GMT 16.65)、30.33%(GMT 17.74)和11.53%(GMT 12.72);0~5岁、6~17岁、18~55岁和56岁4个年龄组中甲型H1N1流感病毒血清抗体阳性率分别为26.88%、30.72%、19.03%和10.38%。 结论 山东省部分人群对甲型H1N1流感病毒抗体水平逐步升高,但仍未达到免疫保护水平。2月山东省甲型H1N1流感抗体阳性率和GMT最高,6~17岁年龄组人群抗体阳性率较其他年龄组高。     

Persistence of pandemic influenza H1N1 virus in young patients after oseltamivir therapy in the 2009–2010 season: a comparison with seasonal H1N1 with or without H275Y mutation

Comparison of the viral persistence of pandemic H1N1 (H1N1pdm) and seasonal H1N1 with or without H275Y mutation after oseltamivir therapy has not been adequately done. Virus was isolated before and on days 4-6 from the start of oseltamivir treatment for 158 cases of seasonal (2007-2008 and 2008-2009 seasons) or pandemic (2009-2010 season) H1N1 influenza. Sequence analysis was done for each season and NA inhibition assay (IC(50)) was done in the 2009-2010 season. H275Y mutation before therapy was 0% in the 2007-2008 and 2009-2010 seasons, but 100% in the 2008-2009 season. Fever and other symptoms were noticeably prolonged after oseltamivir therapy for children with H275Y mutated seasonal H1N1 (2008-2009 season), but not in patients with seasonal H1N1 without mutation (2007-2008) or H1N1pdm (2009-2010). The viral persistence rate was significantly higher for patients 15 years or younger than for those 16 years and older with H275Y mutated seasonal H1N1 (46.2% and 10.5%, respectively) or with H1N1pdm (43.3% and 11.5%, respectively). The H275Y mutation emerged after oseltamivir treatment in 2.4% (2/82) of all patients with H1N1pdm. In two children, the H275Y mutation emerged after therapy and the IC(50) increased more than 200 fold; however, the prolongation of fever was not so prominent. In conclusion, oseltamivir was effective for fever and other clinical symptoms; however, the virus persisted longer than expected after treatment in H1N1pdm influenza-infected children in the 2009-2010 season, similar to seasonal H1N1 with H275Y mutation in the 2008-2009 season.     

Influence of prior pandemic A(H1N1)2009 virus infection on invasion of MDCK cells by community-associated methicillin-resistant Staphylococcus aureus

Secondary bacterial pneumonia due to community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has become a highly publicized cause of death associated with influenza. In this study, we performed the gentamicin-killing assay using Madin-Darby canine kidney (MDCK) cells and MRSA strains to investigate whether prior infection from pandemic A(H1N1)2009 virus (A[H1N1]pdm09) lead to increased invasion of MDCK cells by MRSA. We found that the invasion rate of two MRSA strains (ATCC BAA-1680 [USA 300] and ATCC BAA-1699 [USA 100]) into intact MDCK cell monolayers was 0.29 ± 0.15% and 0.007 ± 0.002%, respectively (p < 0.01, n ≥ 3). In addition, the relative invasion rate of both ATCC BAA-1680 and ATCC BAA-1699 was significantly increased by prior A(H1N1)pdm09 infection of MDCK monolayers from 1 ± 0.28 to 1.38 ± 0.02 and from 1 ± 0.24 to 1.73 ± 0.29, respectively (p < 0.01). These results indicate that ATCC BAA-1680 displays much stronger invasiveness of MDCK cells than ATCC BAA-1699, although invasion of both strains was increased by prior A(H1N1)pdm09 infection. In conclusion, this study provided the first evidence that prior A(H1N1)pdm09 infection facilitates the invasion of MDCK cells by MRSA, presumably due to cellular injury caused by the virus.     

献血人群中SEN病毒感染的检测分析

目的：了解国内献血人群中一种新的比血传播病毒（SENV）感染的流行状况，方法：以SENV ORF1区核苷酸序列设计引物建立套式聚合酶链反应（nPCR）方法。采用多重PCR法对596份严自3个不同地区无偿和／或有偿献血标本进行SENV DNA（D和H亚型）检测，并对PCR阳性产物进行克隆后测序分析。结果：在体检合格的献血中，SENVDNA检出率为13．5％－21．0％在抗－HCV，HBsAg，抗－HIV，梅毒抗体阳性和ALT异常升高的献血中，SENV DNA检出率分别为35．0％、14．0％，60．0％、28．6％和31．3％，献血中SENV－D亚型等高于SENV－H亚型，不同地域献血SENV DNA检出率的差异无显性（P＞0．05），体检不合格献血（抗－HIV或抗－HCV阳性的SENV－D感染率显高于正常献血人群（P＜0．05）；6份严自不同个体和不同地域之间的SENV分离株部分基因组核苷酸的变异最高达11．9％，与标准标（AX025838）相比变异高达13．2％，结论：在国内献血人群中存在SENV感染。     

浙江省德清县农村成人甲型流感各亚型病毒血清抗体检测及其关联探讨

目的 了解浙江省德清县农村社区成人禽流感病毒H7N9、H9、H5、甲型H1N1、季节性H1、H3的血清抗体水平,探索各亚型流感病毒之间的关联。方法 2013年4月德清县发现2例H7N9患者,分别在急性期和恢复期检测了患者及其家属的血清抗体;利用已有的生物样本库,检测病例发生之前收集的725份血清和病例发生之后收集的503份血清的H7N9、H9、H5的抗体水平,了解疫情前后人群血清抗体水平变化情况,在503份血清中抽取398份,检测甲型H1N1、季节性H1、H3的抗体情况,探索其与其他亚型流感病毒的关系;血清检测采用血凝抑制试验(HI),H7N9、H9、H5以HI抗体滴度1: 20为阳性,甲型H1N1和季节性H1、H3以1: 40为阳性,对于部分阳性和阴性样本进行复测;计算抗体阳性率和几何平均滴度,关联分析采用配对2检验并计算关联系数r。结果 H7N9的14名密切接触者中有2人抗体阳性,阳性率为14.3%;H7N9疫情出现之前,一般人群血清中未发现H7N9抗体(0/725),疫情出现之后发现1名隐性感染者,抗体阳性率为0.2%(1/503),H7N9疫情出现前后均未检出H5抗体,H9抗体阳性率没有明显变化,约为4.19%(22/503),标化率4.18%;甲型H1N1阳性率为4.77%(19/398),标化率5.73%;季节性H1为5.28%(21/398),标化率7.86%;季节性H3为11.31%(45/398),标化率6.28%;各个年龄组之间抗体阳性率差异无统计学意义;关联分析显示甲型H1N1血清抗体阳性者,H9血清抗体也趋于阳性(r=0.108, P 0.05);季节性流感病毒H1、H3抗体阳性者,甲型H1N1血清抗体也趋于阳性(r=0.504, P 0.05; r=0.111,P 0.05)。结论 德清县农村社区一般人群对H7N9和H5无免疫力,对H9、甲型H1N1和季节性H3、H1免疫保护水平有限,需要重点监测和保护。甲型H1N1和季节性流感病毒H9可能存在抗体交叉保护或共同暴露。     

Pandemic influenza A (H1N1) in pregnant women: impact of early diagnosis and antiviral treatment

Evaluation of: Siston AM, Rasmussen SA, Honein MA et al. Pandemic 2009 influenza A (H1N1) virus illness among pregnant women in the United States. JAMA 303(15), 1517–1525 (2010).

Pregnant women are at increased risk for severe illness from influenza infection, particularly pandemics, including the current flu pandemic. Early antiviral therapy using oseltamivir or zanamivir may be beneficial, but limited information is available regarding their safety and effectiveness during pregnancy. The article by Siston et al. assessed the findings of a recently published paper describing the severity of illness due to 2009 influenza A (H1N1) infection among 788 pregnant women reported to the CDC, stratified by timing of antiviral therapy and pregnancy trimester at symptom onset. The authors’ findings highlight the potential for severe illness and high risk of mortality due to influenza A (H1N1) infection among pregnant women and suggest the benefit of early antiviral treatment in reducing intensive care unit admissions and mortality rate. The WHO and CDC recommendations including 2009 influenza A (H1N1) vaccination and early antiviral therapy in case of influenza-like illness for all pregnant women are discussed regarding the key findings of this paper and other published data on influenza A (H1N1) infection in pregnant women.