反向聚合酶链反应检测肝细胞癌中乙型肝炎病毒DNA整合

目的应用反向聚合酶链反应(inverse polymerase chain reaction,IPCR)法检测肝细胞癌(HCC)中乙型肝炎病毒(HBV)DNA整合.方法提取手术切除石蜡包埋人肝癌组织中DNA,根据IPCR原理,选用HBV DR1区无酶切位点的限制性内切酶酶切,通过连接反应形成环化DNA分子,以此作为模板进行PCR扩增得到已知序列的旁侧序列.琼脂糖凝胶电泳观察PCR扩增产物片段.结果 19例HCC标本中有14例检出整合型HBV DNA,2例同时存在游离型HBV DNA.结论 IPCR可以准确测定肝细胞中HBV DNA的整合.该方法为研究HBV DNA在肝细胞中的整合机制提供一简单、快速、经济途径.     

应用巢式PCR扩增孕妇血浆中胎儿SRY基因的研究

目的:建立一种巢式聚合酶链反应(PCR),利用孕妇血浆中的游离胎儿DNA(fetal DNA)鉴定胎儿SRY基因。方法:随机采集30例孕妇外周血标本,采用酚/氯仿法从孕妇血浆中提取DNA,设计两对引物对SRY基因进行巢式PCR扩增,扩增产物经测序加以确认。结果:17例孕男胎的孕妇血浆中有15例经巢式PCR扩增检出SRY基因,而13例孕女胎的孕妇血浆没有检出阳性结果。准确性和敏感性分别为93.3%(28/30)和88.2%(15/17)。结论:应用酚/氯仿法从孕妇血浆中提取游离DNA简单有效,结合巢式PCR扩增SRY基因技术可用于无创性产前性连锁遗传性疾病的诊断。     

男性乙肝大三阳携带者精浆乙肝病毒核酸的检测

目的 探讨乙肝大三阳携带者精浆乙肝病毒核酸(HBV DNA)的检测方法、含量及其影响因素.方法 选择男性乙肝大三阳携带者99例,根据血清HBV DNA定量级别分为107组和108组,采用沉淀煮沸裂解法(实验组)和微量DNA提取法(DNA Mini法)(对照组)抽提精浆的HBV DNA模板,进行荧光定量PCR检测.结果 实验组和对照组检测精浆HBV DNA的阳性率分别为62.0％和61.2％,两者比较差异无统计学意义(P＞0.05).经2种DNA抽提法检测的10 7组和10 8组精浆HBV DNA的阳性率分别为35.3％和89.6％,差异有统计学意义(P＜0.01).经对数转换后,血清与精浆HBV DNA含量分别为(7.98±0.42)IU/ml和(3.05±1.58)IU/ml,差异有统计学意义(P＜0.01).结论 沉淀煮沸裂解法能有效抽提精浆的HBV DNA进行荧光定量PCR测定,乙肝大三阳携带者精浆HBV DNA含量低于血清HBVDNA含量,精浆HBV DNA的阳性率与血清HBV DNA水平密切相关.     

细菌通用引物PCR快速诊断全身性感染的研究

目的:建立PCR直接检测临床血标本中病原菌的方法,探索其快速诊断全身性感染(sepsis)的应用价值.方法:利用16S rRNA基因的高度保守性,设计并合成细菌的通用引物;采用合成的引物扩增标准菌株及全身性感染病人的血样本,并与血培养进行对照.结果:通用引物PCR可检出0.005pg的大肠埃希菌标准菌株DNA;临床分离的大肠埃希菌、金黄色葡萄球菌PCR扩增产物于255bp处均可见清晰电泳条带,正常人无菌血白细胞及白色念珠菌DNA扩增产物均未见电泳条带出现;PCR可快速地检出血液中多种病原菌DNA,所检的8例病人中4例阳性(50%).结论:165 rRNA基因为基础的PCR,可直接用于临床血标本病原菌的检测,初步实验显示其具有特异、快速等优点,敏感性较血培养有增高趋势,有利于重症急性胰腺炎及其他外科危重病Sepsis的早期诊断.     

外周血单个核细胞中HBV DNA与HBV cccDNA对慢性乙型肝炎患者免疫功能的影响

目的 比较外周血单个核细胞(PBMCs)有或无HBV感染证据组中免疫功能的差异.方法 常规分离78例CHB患者和10例健康对照组全血中的PBMCs,提取DNA,应用实时荧光定量聚合酶链反应(PCR)法检测PBMCs中的HBV DNA和HBV cccDNA;进行PBMCs的培养,孵育72 h后,收集PBMCs上清液,采用酶联免疫吸附法(ELISA)检测干扰素-γ(IFN-γ)和白细胞介素-10(IL-10),并用四甲基偶氮唑盐(MTT)法检测PBMCs的增殖活性.结果 PBMCs中HBV cccDNA阳性的患者组,其PBMCs产生IFNγ水平显著下降,IL-10水平显著升高,同时伴PBMCs的增殖能力显著下降.结论 HBV感染PBMCs可能是HBV感染后机体Th1/Th2细胞因子比例失衡,特异性免疫功能低下,形成慢性HBV感染的主要原因之一.     

PCR-RFLP检测维生素D受体基因启动子区多态性

目的建立一种简便、实用、重复性好的人维生素D受体（vitamin D receptor，VDR）基因5’端启动子区Fokl酶切位点多态性的检测方法，以利于VDR基因起始密码多态性与骨质疏松等疾病相关性的研究。方法应用聚合酶链反应（PCR）特异性扩增VDR基因5’端启动子区，扩增产物用限制性内切酶Fokl酶切，10％聚丙酰胺凝胶电泳观察酶切结果。结果应用PCR—RFLP法检测了98例健康汉族中老年人VDR基因型，FF、Ff、ff基因型分布频率分别为47％、43％、10％。结论该方法简便、准确、实用，适用于院校及医院实验室的研究和分子流行病学调查。     

乙型肝炎后肝硬化肝移植乙型肝炎病毒再感染检测

目的探讨乙型肝炎病毒（HBV）DNA、YMDD变异及血清标志物在乙型肝炎肝硬化患者肝移植后HBV再感染检测的临床意义。方法定量聚合酶链反应（PCR）检测HBV DNA；HBV PCR产物限制性片段长度多态性分析检测YMDD变异；酶联免疫检测HBV血清标志物；分析乙型肝炎患者肝移植后YMDD变异、HBV DNA及血清标志物检测结果。结果乙型肝炎肝硬化患者肝移植术后HBV再感染率为8．9％（17／189）；YMDD变异病例达HBV再感染病例58．8％（10／17）；HBV DNA定量检测有较高的灵敏度，能早期诊断HBV的再感染，有利于临床治疗；对HBV DNA阴性但HBsAg阳性病例，PreS1和HBeAg血清标志物可用于病毒复制的补充检测。结论乙型肝炎肝硬化患者肝移植后预防乙型肝炎复发应采用HBV DNA、YMDD变异及血清标志物联合检测。     

PBMCs HBV逆转录酶mRNA表达与乙型肝炎的临床关系

目的探讨乙型肝炎病毒（HBV）逆转录酶mRNA存外周血单个核细胞（PBMCs）中的表达与乙型肝炎的临床关系。方法选择70例乙型肝炎患者（包括急性乙型肝炎、慢性乙型肝炎、肝硬化）及10例HBsAg阴性的健康体检者。分别提取其PBMCs中的总RNA,经无RNA酶的DNA酶I消化去除残存的基因组DNA后,逆转录合成cDNA。以HBVP基因区设计合成逆转录酶引物进行PCR扩增,检测HBV逆转录酶的表达情况。对1例阳性标本进行了克隆和测序,将所得序列与Gen．Bank数据库中已知核苷酸序列进行同源性分析。结果所得基因序列为379个核苷酸,与实验设计相符。与HBVadw2亚型标准一级结构比较,同源性为93．7％。对照组无1例HBV逆转录酶mRNA阳性表达;急性乙型肝炎组阳性表达率较低,为5．3％（1／15）;慢性乙型肝炎组阳性表达率为33．3％（12／36）,两组阳性率差异有显著统计学意义（P〈0．05）;肝硬化组阳性表达率为26．7％（4／15）,与慢性乙型肝炎组比较,阳性率差异无统计学意义（P〉0．05）。PBMCs中HBV逆转录酶mRNA表达阳性组与阴性组肝功能比较,大部分指标差异有显著统计学意义（P〈0．05）。PBMCs中HBV逆转录酶表达阳性率与血清HBeAg、HBVDNA阳性率之间无相关性（r=0．164、,=0．07）,其差异均无显著统计学意义（P〉0．05）。另外,乙型肝炎患者中,有家族史者的PBMCs逆转录酶mRNA阳性表达率占37．0％（10／27）,而无家族史者的仅占16．3％（7／43）,乙型肝炎患者的家族史与PBMCsHBV逆转录酶阳性表达有相关性（r=0．229）,差异有显著统计学意义（P〈0．05）。结论HBV逆转录酶在PBMCs中可独立复制,其表达与乙型肝炎的慢性化、重度化及家族聚集性有密切相关性,可为早期诊断指标之一。     

核苷(酸)类药物治疗慢性乙型肝炎的长期性

<正>近20年来,临床和流行病学研究表明,持续、活跃的乙型肝炎病毒(HBV)复制是慢性乙型肝炎(CHB)疾病进展的最重要因素,只有长期抑制HBV复制,才有可能防止疾病进展。但HBV在复制过程中,在肝细胞核内形成共价闭合环状DNA(cccDNA)并可持续存在~([1]);核苷(酸)类药物(NAs)主要抑制HBV复制的逆转录环节,对cccDNA无直接抑制或清除作用;CHB患者存在特异性免疫功能障碍~([2-5]),而NAs无直接的免疫调节作用~([5])。因此,现有NAs抗病毒治疗很难彻底清除HBV,仅部分HBV e抗原     

慢性乙型肝炎患者肝组织 HBV cccDNA含量与血清 HBV 标志物和病毒载量的相关性

乙型肝炎病毒共价闭合环状 DNA （ HBV cccDNA）是嗜肝病毒持续感染和HBV复制的突出标志,也是评价HBV感染状态及治疗效果最客观、敏感的指标。监测肝脏内 HBV cccDNA的含量对于病情判断、疗效和预后评估均具有重要意义。本研究通过检测拉米夫定初治后患者肝组织内HBV cccDNA 的含量及血清 HBsAg、HBeAg、HBV DNA等水平,探讨肝组织内HBV cccDNA的含量与HBV血清学标志物和病毒载量的相关性。     

Change of Hepatitis B Virus DNA and Covalently Closed Circular DNA Status in Liver Graft After Liver Transplantation

Recurrence of hepatitis B virus (HBV) occurs despite prophylaxis, and covalently closed circular DNA (cccDNA) is thought to play a role owing to its resistance to prophylactic agents used. The aim of this study was to evaluate the changes of HBV DNA and cccDNA within the liver graft during liver transplantation (LT). Polymerase chain reaction (PCR) primers and probes were designed to measure total HBV DNA (tDNA) and cccDNA by real-time PCR. One hundred fifty samples from 70 patients who underwent LT for HBV were used for analysis. A 1st biopsy was taken from the donor before donor hepatectomy (Bx1), a 2nd from the recipient after reperfusion (Bx2), and a 3rd (Bx3) during follow-up after LT in 18 patients. Both tDNA and cccDNA after reperfusion were detected more frequently in pre-LT HBeAg⁺ and high–HBV DNA titer recipients. However, the type and duration of antiviral agents and presence of mutation before LT did not influence the presence of tDNA or cccDNA in Bx2. tDNA positivity within the graft decreased from 41.4% to 22.2% during follow-up, but cccDNA did not (4.3% in Bx2 and 5.6% in Bx3). Although HBV recurrence was not related to pre-LT recipient HBeAg or HBV DNA titer, the presence of tDNA after reperfusion had strong correlation. The presence of tDNA within the graft is influenced by pre-LT viral replicative status, and although its presence decreases with prophylaxis, it is strongly correlated with recurrence. cccDNA does not have a role in predicting recurrence but is preserved within the graft despite prophylaxis.     

Impact of Intrahepatic Hepatitis B DNA and Covalently Closed Circular DNA on Survival After Hepatectomy in HBV-Associated Hepatocellular Carcinoma Patients

Background

Chronic hepatitis B virus (HBV) infection induces persistent but ineffective immune activation that contributes to necroinflammation, fibrosis, and risk of hepatocellular carcinoma (HCC). This study aims to determine the relationship of intrahepatic total HBV (ih HBV) DNA and covalently closed circular DNA (cccDNA) with necroinflammation and fibrosis, and their impact on prognosis after resection in HBV HCC patients.

Methods

Data are from 111 patients treated with primary liver resection for HBV HCC at Mount Sinai, New York (1991–2008). ih HBV DNA and cccDNA were quantitated by real-time PCR. Necroinflammation was graded according to histologic activity index (HAI), and liver fibrosis was staged by the modified Ishak method.

Results

A total of 106 (95 %) and 89 patients (80 %) had detectable ih HBV DNA and cccDNA, respectively, while 43 % had detectable serum HBV DNA. cccDNA made up a small proportion of ih HBV DNA (median cccDNA/ih HBV DNA ratio = 0.022). Higher levels of ih HBV DNA were associated with higher HAI and serum alanine aminotransferase (ALT), while a lower ratio of cccDNA/ih HBV DNA was associated with higher HAI, ALT, and Ishak fibrosis stage. ih HBV and cccDNA were not associated with survival, but the lowest quintile of cccDNA/ih HBV DNA ratio (<0.0024) was independently associated with poor overall survival.

Conclusions

A lower cccDNA/ih HBV DNA ratio was associated with greater necroinflammation and liver fibrosis, and was independently associated with poor overall survival. Thus, intracellular virus loads and relative proportions of virus DNA reflect histologic damage in the liver and influence the clinical outcome of HBV HCC patients.     

Correlation Between Risk of Hepatitis B Virus Recurrence and Tissue Expression of Covalently Closed Circular DNA in Living Donor Liver Transplant Recipients Treated With High-dose Hepatitis B Immunoglobulin

Background and AimsDespite the application of prophylaxis, the risk of hepatitis B virus (HBV) recurrence remains. However, actual mechanism(s) and definite risk factor(s) are obscure. The present study examined the correlation between the HBV load in liver explants and post-liver transplant (OLT) HBV recurrence.MethodsHBV DNA was extracted from liver tissue taken from 50 living donor OLT (LDLT) patients using the QuickGene DNA Tissue Kit S (Fujifilm, Tokyo, Japan) and subjected to real-time polymerase chain reaction with the following primers: 5′-CACATGGCCTCCAAGGAGTAA-3′ (forward primer) and 5′-TGAGGGTCTCTCTCTTCCTCTTGT-3′ (reverse primer). To prevent HBV infection, patients were treated daily with high-dose (10,000 IU) hepatitis B immunoglobulin (HBIG) for the first week after LDLT. They then received weekly doses for the next month and then monthly doses for ≤1 year. If the anti-hepatitis surface antigen antibody titer was <1,000 IU/L, an antiviral agent (AVA) was added to the regimen.ResultsThe mean (±SD) tissue HBV DNA and covalently closed circular DNA (cccDNA) loads were ?0.8 ± 1.2 (range, ?2.9 to 2.6) and ?2.3 ± 1.1 (range, ?4.6 to 0.6) log₁₀ copies/cell, respectively. There was a significant correlation between serum and tissue HBV DNA (r = 0.65; P = .00) and cccDNA concentrations (r = 0.55; P = .00). Six patients suffered HBV recurrence and 9 required additional AVA. There was no direct correlation between HBV recurrence and tissue cccDNA concentration. However, the concentration of cccDNA was significantly greater those patients suffering recurrence and receiving AVA treatment (high-risk group).ConclusionHigh tissue cccDNA concentrations may be a risk factor for HBV recurrence despite high-dose HBIG prophylaxis.     

Visual detection of porcine reproductive and respiratory syndrome virus using CRISPR‐Cas13a

Porcine reproductive and respiratory syndrome virus (PRRSV) has varied constantly and circulated in the pig industry worldwide. The prevention and control of porcine reproductive and respiratory syndrome (PRRS) is complicated. A visual, sensitive and specific diagnostic method is advantageous to the control of PRRS. The collateral cleavage activity of LwCas13a is activated to degrade non‐targeted RNA, when crRNA of LwCas13a bond to target RNA. The enhanced Cas13a detection is the combination of collateral cleavage activity of LwCas13a and recombinase polymerase amplification (RPA). In this study, the enhanced Cas13a detection for PRRSV was established. The novel method was an isothermal detection at 37°C, and the detection can be used for real‐time analysis or visual readout. The detection limit of the enhanced Cas13a detection was 172 copies/μl, and there were no cross‐reactions with porcine circovirus 2, porcine parvovirus, classical swine fever virus and pseudorabies virus. The enhanced Cas13a detection can work well in clinical samples. In summary, a visual, sensitive and specific nucleic acid detection method based on CRISPR‐Cas13a was developed for PRRSV.     

甲泼尼龙和他克莫司在体外对HepG2.2.15细胞中乙型肝炎病毒cccDNA合成的影响

目的 探讨甲泼尼龙(MP)和他克莫司(Tac)在体外对乙型肝炎病毒(HBV)cccDNA合成的影响.方法 采用 HepG2.2.15细胞(HBV永久转染人肝癌细胞系)为体外实验模型,经四甲基偶氮唑盐(MTT)试验确定MP的安全浓度在0～250 ng/ml,Tac的安全浓度在O～500 ng/ml.将不同安全浓度的 MP(O、10、50、100及250 ng/ml)和Tac(0、50、100及500 ng/ml)分别作用于该细胞系,72 h收集细胞培养上清液及细胞,采用实时定量聚合酶链反应检测上清液中HBV DNA及细胞内HBV cccDNA水平.结果 与不含MP者比较,当MP浓度分别为10、50、100及250 μg/L时,HBV DNA和HBV cccDNA水平明显降低,分别为(5.7823±0.1861)、(5.1337±0.1364)、(4.7865± 0.0398)及(4.1468±0.1016)log_(10)拷贝/ml(P<0.05,P0.05).结论 在体外,MP能抑制HepG2.2.15细胞中HBV DNA的表达,同时可以减少细胞内HBV cccDNA的合成,该作用呈剂量依赖性;Tac无此作用.     

复方肃毒星提取物抑制乙型肝炎病毒cccDNA的作用

目的研究复方肃毒星提取物（简称肃毒星）体外对乙型肝炎病毒（HBV）复制模板共价闭合环状DNA（cccDNA）的抑制作用。 方法通过细胞计数试剂盒CCK-8检测肃毒星在恩替卡韦耐药HBV稳定复制细胞系（HepG2.A64）、野生HBV稳定复制细胞系（HepG2.2.15和HepAD38）以及肝癌细胞系（HepG2）中的毒性作用；选择安全有效的高、中、低浓度肃毒星分别处理3个细胞系，采用实时荧光定量PCR法检测细胞内HBV cccDNA水平。以高斯荧光素酶微环cccDNA（mc-cccDNA）转染HepG2细胞，同时选择高、中、低浓度的肃毒星在不同时间点作用转染mc-cccDNA的HepG2细胞，荧光素酶检测试剂盒分析相对荧光强度，计算对cccDNA的抑制率，评价cccDNA转录活性。 结果肃毒星在HepG2.A64、HepG2.2.15、HepAD38和HepG2细胞中的半数毒性浓度分别为27.01 μg/ml、29.36 μg/ml、31.20 μg/ml和52.80 μg/ml。肃毒星对HepG2.A64、HepG2.2.15和HepAD38细胞中HBV cccDNA有抑制作用，最佳作用时间为5 d，5 d最大抑制率分别为（84.24 ± 2.1）%、（52.02 ± 4.74）%和（47.16 ± 6.69）%，与未用药处理组差异有统计学意义（P均< 0.05）。肃毒星对转染HepG2细胞后mc-cccDNA转录活性的最佳抑制率为（48.44 ± 4.54）%，抑制作用优于干扰素对照组（P < 0.05），略弱于特异性敲低mc-cccDNA的pLV-CRISPR处理对照组（P < 0.05），差异有统计学意义。 结论肃毒星对恩替卡韦耐药型和野生型细胞系HBV cccDNA的复制及转录均有很好的抑制作用，为慢性乙型肝炎功能学治愈新药的开发提供参考。     

高发区乙型肝炎病毒致肝细胞癌风险的病毒学特征

目的 研究启东高发区人群慢性乙型肝炎病毒（HBV）HBeAg、共价闭环DNA（CCCDNA）的表达和HBV核苷酸1762／1764错义突变与肝细胞癌发生的关系。方法 自1989年1月至2002年12月,随访启东地区377例临床诊断为慢性HBV感染的中年男性,选取发生肝癌的32例患者,与年龄、职业、生活环境相匹配的慢性HBV感染患者32例组成巢式病例对照研究队列。采用ELISA法检测血清HBeAg表达情况,半巢式PCR扩增HBVcccDNA,巢式PCR扩增HBVX基因并测序,分析核苷酸1762／1764点突变,比较肝癌组和对照组在HBV风险特征上的差异。结果肝癌组和对照组血清HBeAg的阳性率分别是53．1％（17／32）和15．6％（5／32）（P〈0．01）;HBVcccDNA的阳性率分别是62．5％（21／32）和25．0％（8／32）（P〈0．01）;HBV核苷酸1762／1764的突变率分别为82．1％（23／28）和71．4％（20／28）（P=0．425）。结论 HBeAg和CCCDNA所反映的HBV滴度和活跃复制状态与肝癌的发生密切关联,可作为预测肝癌发生的风险指标。     

Serological and vaccination profile of hemodialysis patients during an outbreak of hepatitis B virus infection

During an outbreak of hepatitis B virus (HBV) infection in a hemodialysis unit, patients were assessed for serological viral markers and vaccination status. HBV infection was identified in 26 patients. Twenty of these were positive for hepatitis B surface antigen (HBsAg), and 6 were negative for HBsAg but positive for IgM antibody to hepatitis B core (anti-HBc) and HBV DNA. The primary source of infection was not clearly identified, although 2 patients were suspected to be the index cases. A multiple logistic regression analysis revealed low anti-HBs titers and vaccination status to be independently associated with the risk of acquiring HBV infection. Both the high prevalence of HBV infection (31%) detected in this unit and the low vaccine response (53%) observed reinforce the importance of universal and preventive measures in controlling HBV infection. The detection of HBV DNA in HBsAg-negative/IgM anti-HBc-positive patients emphasizes the value of anti-HBc testing in the routine screening of HBV in hemodialysis units.     

Absence of hepatitis B and C viruses in pediatric idiopathic membranoproliferative glomerulonephritis

The blood-borne hepatitis viruses, hepatitis B virus (HBV) and hepatitis C virus (HCV), have similar epidemiological features. The association of chronic HBV infection and glomerulonephritis is well established, particularly in children. Recent reports have shown an association between HCV infection and glomerulonephritis in adults. In order to assess the role of these hepatotropic viruses in membranoproliferative glomerulonephritis (MPGN) we screened 34 children with idiopathic MPGN for the presence of HBV and HCV infection using highly sensitive polymerase chain reaction techniques for the detection of HBV DNA and HCV RNA. Also, enzyme-linked immunosorbent assays were used to detect the presence of antibody to hepatitis B surface antigen and antibody to HCV. No evidence of HBV or HCV infection was demonstrated in any of the patients. We conclude that HBV and HCV are not significant causes of idiopathic MPGN in children in the United States.     

四种HBV DNA 荧光定量PCR试剂比较及其结果分析

目的 评价3种国产HBV DNA荧光定量PCR试剂的特异性和灵敏度.方法 采用德国QIAGEN公司的Artus HBV DNA荧光定量PCR试剂(A)和国内生产的3种HBV DNA荧光定量PCR试剂(B、C、D)对203例HBV感染者血清标本进行平行检测,分析3种国产试剂与Artus试剂检测结果的差别及其相关性.结果 4种试剂检测203例血清标本HBV DNA的结果分别是A (5.78±1.78)、B(5.70±1.34)、C(5.51±1.96)、D(5.60±1.52),3种试剂与Artus试剂的相关系数分别为B:0.860、C:0.954、D:0.922. 对于HBV DNA载量高于106拷贝/ml的标本,3种国产试剂检测结果的均值与Artus无明显差异,但试剂B与Artus相关性明显不如试剂C和试剂D;对于HBV DNA载量在103～106 拷贝/ml的标本,试剂B和试剂C的均值水平与Artus试剂存在明显差别,试剂C和D与Artus试剂的相关性要优于试剂B;而对于30～103 拷贝/ml的标本,试剂B和试剂D的均值与Artus结果存在显著差异,且3种国产试剂与Artus试剂相关性很差.在26例30～103拷贝/ml样本中,3种国产试剂分别漏检8例(30.8%)、8例(30.8%)和11例(42.3%),7例<30拷贝/ml样本中3种国产试剂无一检出.结论 3种国产试剂的敏感性均较Artus试剂差,尤其是病毒载量低于103拷贝/ml的标本.在3种国产试剂中,试剂C与Artus试剂检测的结果相关性最好.