Viral diseases represent one of the major threats for salmonids aquaculture.Early detection and identification of viral pathogens is the main prerequisite prior to undertaking effective prevention and control measures...Viral diseases represent one of the major threats for salmonids aquaculture.Early detection and identification of viral pathogens is the main prerequisite prior to undertaking effective prevention and control measures.Rapid,sensitive,efficient and portable detection method is highly essential for fish viral diseases detection.Biosensor strategies are highly prevalent and fulfill the expanding demands of on-site detection with fast response,cost-effectiveness,high sensitivity,and selectivity.With the development of material science,the nucleic acid biosensors fabricated by semiconductor have shown great potential in rapid and early detection or screening for diseases at salmonids fisheries.This paper reviews the current detection development of salmonids viral diseases.The present limitations and challenges of salmonids virus diseases surveillance and early detection are presented.Novel nucleic acid semiconductor biosensors are briefly reviewed.The perspective and potential application of biosensors in the on-site detection of salmonids diseases are discussed.展开更多
Screening tests for blood donations are based upon sensitivity, cost-effectiveness and their suitability for high-throughput testing. Enzyme immunoassay (EIAs) for hepatitis C virus (HCV) antibodies were the initial s...Screening tests for blood donations are based upon sensitivity, cost-effectiveness and their suitability for high-throughput testing. Enzyme immunoassay (EIAs) for hepatitis C virus (HCV) antibodies were the initial screening tests introduced. The ”first generation“ antibody EIAs detected seroconversion after unduly long infectious window period. Improved HCV antibody assays still had an infectious window period around 66 d. HCV core antigen EIAs shortened the window period considerably, but high costs did not lead to widespread acceptance. A fourth-generation HCV antigen and antibody assay (combination EIA) is more convenient as two infectious markers of HCV are detected in the same assay. Molecular testing for HCV-RNA utilizing nucleic acid amplification technology (NAT) is the most sensitive assay and shortens the window period to only 4 d. Implementation of NAT in many developed countries around the world has resulted in dramatic reductions in transfusion transmissible HCV and relative risk is now < 1 per million donations. However, HCV serology still continues to be retained as some donations are serology positive but NAT negative. In resource constrained countries HCV screening is highly variable, depending upon infrastructure, trained manpower and financial resource. Rapid tests which do not require instrumentation and are simple to perform are used in many small and remotely located blood centres. The sensitivity as compared to EIAs is less and wherever feasible HCV antibody EIAs are most frequently used screening assays. Efforts have been made to implement combined antigen-antibody assays and even NAT in some of these countries.展开更多
BACKGROUND Rapid molecular testing has revolutionized the management of suspected viral meningitis and encephalitis by providing an etiological diagnosis in<90 min with potential to improve outcomes and shorten inp...BACKGROUND Rapid molecular testing has revolutionized the management of suspected viral meningitis and encephalitis by providing an etiological diagnosis in<90 min with potential to improve outcomes and shorten inpatient stays.However,use of molecular assays can vary widely.AIM To evaluate current practice for molecular testing of pediatric cerebrospinal fluid(CSF)samples across the United Kingdom using a structured questionnaire.METHODS A structured telephone questionnaire survey was conducted between July and August 2020.Data was collected on the availability of viral CSF nucleic acid amplification testing(NAAT),criteria used for testing and turnaround times including the impact of the coronavirus disease 2019 pandemic.RESULTS Of 196/212(92%)microbiology laboratories responded;63/196(32%)were excluded from final analysis as they had no on-site microbiology laboratory and outsourced their samples.Of 133 Laboratories included in the study,47/133(35%)had onsite facilities for viral CSF NAAT.Hospitals currently undertaking onsite NAAT(n=47)had much faster turnaround times with 39 centers(83%)providing results in≤24 h as compared to those referring samples to neighboring laboratories(5/86;6%).CONCLUSION Onsite/near-patient rapid NAAT(including polymerase chain reaction)is recommended wherever possible to optimize patient management in the acute setting.展开更多
目的基于血液筛查核酸检测反应性献血者的HBV感染的确认,探讨核酸检测反应性献血者的归队策略。方法联合应用自建的高灵敏度核酸检测体系、血液核酸筛查等多种核酸检测(NAT)方法,并结合血清学检测、献血者随访,对核酸检测反应性(NAT-yie...目的基于血液筛查核酸检测反应性献血者的HBV感染的确认,探讨核酸检测反应性献血者的归队策略。方法联合应用自建的高灵敏度核酸检测体系、血液核酸筛查等多种核酸检测(NAT)方法,并结合血清学检测、献血者随访,对核酸检测反应性(NAT-yield)献血者中的HBV感染进行确认和感染状态识别。依据确认的HBV感染血浆样本,比较不同确认方法、确认指标或指标组合对HBV感染确认的效果。结果2010年11月—2021年2月,在血液筛查检出的876位NAT-yield献血者中共确认HBV感染者511人(OBI 451人,急性早期HBV感染者27人,不能确认感染者33人,无感染者30人,不能确认HBV感染者335人)。采用单检系统对混检系统检出的HBV感染血浆进行复测的检出率为96.6%,明显高于混检系统对单检系统检出的HBV DNA反应性(HBV DNA R)组和鉴别试验无反应性(NDR)组的复测检出率(76.4%和55.7%)(P<0.05)。NDR样本在模式2(ID×5+鉴别×2)下复测检出率(65.2%)高于模式1(ID×2+鉴别×1)(39.2%)(P<0.05);2种单检复测模式下的HBV DNA R样本复测检出率无明显差异(P>0.05),但均明显高于NDR样本(P<0.05)。回溯OBI献血者既往NAT数据,有46%经历多次NAT检测而未能检出。有59.1%OBI献血者随访检不出HBV DNA。OBI献血者中抗-HBc+占比为90.2%,单独抗-HBc+为49.2%,远高于不能确认感染组(P<0.05);HBeAg、抗-HBe和抗-HBc IgM在OBI和不能确认感染组中的比例极低且无差异(P>0.05)。结论近60%的NAT-yield献血者可以确认HBV感染。为保证献血者归队的安全性,需要更高灵敏度的HBV DNA确证技术提高HBV感染的确认率。抗-HBc是NAT-yield献血者OBI风险排查和归队评估最重要的血清学指标。展开更多
目的通过危害分析及关键控制点(hazard analysis and critical control point,HACCP)体系系统化控制核酸检测实验室质量,以期能够实现核酸检测实验室质量的提升,为有效制定核酸检测质量控制对策提供借鉴。方法根据HACCP体系初步探究核...目的通过危害分析及关键控制点(hazard analysis and critical control point,HACCP)体系系统化控制核酸检测实验室质量,以期能够实现核酸检测实验室质量的提升,为有效制定核酸检测质量控制对策提供借鉴。方法根据HACCP体系初步探究核酸检测实验室质量控制,重点分析HACCP体系在核酸标本接收、准备试剂、提取标本核酸、核酸扩增、结果判读、核酸检测结果报告环节制定的监控程序、验证措施以及纠偏措施等重要控制点。结果通过分析2021年12月-2022年12月的检测数据,结果显示,各批次核酸检测阳性质控品均满足判定规则,每一批次的实验均处于在控状态。对HACCP体系下实验室数据进行统计,计算其标准差、均值以及变异系数,对箱型图绘制后,展开离群值检验,结果发现,均值没有产生离群值。结论核酸检测实验室中应用HACCP体系,能够有效控制核酸检测实验室质量,全面掌控核酸检测实验室重要控制点,实现实验室生物安全水平与质量的提升,保证核酸检测实验室质控的有效性与科学性,继而有效控制核酸检测实验室质量。展开更多
目的通过分析罗氏Cobas s 201的血液核酸检测(NAT)结果评估其对HBV的检测效果。方法将检测结果根据酶联免疫吸附试验(ELISA)和NAT混合检测(MP)、NAT单样本检测(ID)以及重复NAT单样本检测(rID)分组,分为ELISA+/NAT(ID)+、ELISA+/NAT(rID)...目的通过分析罗氏Cobas s 201的血液核酸检测(NAT)结果评估其对HBV的检测效果。方法将检测结果根据酶联免疫吸附试验(ELISA)和NAT混合检测(MP)、NAT单样本检测(ID)以及重复NAT单样本检测(rID)分组,分为ELISA+/NAT(ID)+、ELISA+/NAT(rID)+、ELISA-/NAT(ID)+、ELISA-/NAT(rID)+4组进行统计分析,探讨重复NAT对反应性结果的检出是否存在差异,对于不同ELISA结果的NAT反应性标本的循环阈值(cycle threshold,Ct)与核酸检出率的关联性。再通过补充试验,包括其他方法学的NAT系统和化学发光血清学标志物检测,进一步分析献血者的真实感染情况。结果766293份献血者标本中共有1691组HBV NAT(MP)+,其中1418组(83.86%)检出反应性结果(1418份HBV NAT+,7090份NAT-),仍有273组(16.14%)经重复检测仍未检出[共计1638份NAT-,Ct(MP):39.49±3.62]。HBV NAT+中,881份(62.13%)ELISA+/NAT(ID)+,19份(1.34%)ELISA+/NAT(rID)+,451份(31.81%)ELISA-/NAT(ID)+,67份(4.72%)ELISA-/NAT(rID)+。对于不同ELISA结果的标本,重复NAT对HBV的检出存在差异(P<0.05)。各组间Ct(ID)值仅ELISA+/NAT(rID)+和ELISA-/NAT(ID)+、ELISA+/NAT(rID)+和ELISA-/NAT(rID)+2组比较无差异(P>0.05),其余各组间两两比较,均有差异(P<0.05)。对228份ELISA-/NAT(MP)+(ID)-进行补充试验,有56份(24.56%)经化学发光检测HBsAg+和7份(3.07%)经其他NAT系统检出反应性。剩余221份(96.93%)NAT-标本中,53份(23.98%)HBsAg+的献血者可能存在慢性感染,40份(18.10%)抗-HBe+和(或)抗-HBc+的献血者可能存在既往感染,其余128份(57.92%)均无反应性的献血者为NAT(MP)假反应性,且各组间抗-HBs含量差异较大(P<0.05)。结论重复NAT对不同反应性类别或不同血清学结果的献血者标本存在差异性检出,尤其在一定区间范围内,对于ELISA-标本进行重复NAT可明显提高检出率。Ct值可辅助评估NAT系统的稳定性和准确性。对于ELISA-/NAT(MP)+(ID)-献血者,结合其他高灵敏度的检测手段可降低病毒残余风险,保障临床用血安全。展开更多
基金supported by the National Key Research and Development Program of China(2022YFC2601304)National Key Research and Development Program of China(2022YFC2602100)。
文摘Viral diseases represent one of the major threats for salmonids aquaculture.Early detection and identification of viral pathogens is the main prerequisite prior to undertaking effective prevention and control measures.Rapid,sensitive,efficient and portable detection method is highly essential for fish viral diseases detection.Biosensor strategies are highly prevalent and fulfill the expanding demands of on-site detection with fast response,cost-effectiveness,high sensitivity,and selectivity.With the development of material science,the nucleic acid biosensors fabricated by semiconductor have shown great potential in rapid and early detection or screening for diseases at salmonids fisheries.This paper reviews the current detection development of salmonids viral diseases.The present limitations and challenges of salmonids virus diseases surveillance and early detection are presented.Novel nucleic acid semiconductor biosensors are briefly reviewed.The perspective and potential application of biosensors in the on-site detection of salmonids diseases are discussed.
文摘Screening tests for blood donations are based upon sensitivity, cost-effectiveness and their suitability for high-throughput testing. Enzyme immunoassay (EIAs) for hepatitis C virus (HCV) antibodies were the initial screening tests introduced. The ”first generation“ antibody EIAs detected seroconversion after unduly long infectious window period. Improved HCV antibody assays still had an infectious window period around 66 d. HCV core antigen EIAs shortened the window period considerably, but high costs did not lead to widespread acceptance. A fourth-generation HCV antigen and antibody assay (combination EIA) is more convenient as two infectious markers of HCV are detected in the same assay. Molecular testing for HCV-RNA utilizing nucleic acid amplification technology (NAT) is the most sensitive assay and shortens the window period to only 4 d. Implementation of NAT in many developed countries around the world has resulted in dramatic reductions in transfusion transmissible HCV and relative risk is now < 1 per million donations. However, HCV serology still continues to be retained as some donations are serology positive but NAT negative. In resource constrained countries HCV screening is highly variable, depending upon infrastructure, trained manpower and financial resource. Rapid tests which do not require instrumentation and are simple to perform are used in many small and remotely located blood centres. The sensitivity as compared to EIAs is less and wherever feasible HCV antibody EIAs are most frequently used screening assays. Efforts have been made to implement combined antigen-antibody assays and even NAT in some of these countries.
文摘BACKGROUND Rapid molecular testing has revolutionized the management of suspected viral meningitis and encephalitis by providing an etiological diagnosis in<90 min with potential to improve outcomes and shorten inpatient stays.However,use of molecular assays can vary widely.AIM To evaluate current practice for molecular testing of pediatric cerebrospinal fluid(CSF)samples across the United Kingdom using a structured questionnaire.METHODS A structured telephone questionnaire survey was conducted between July and August 2020.Data was collected on the availability of viral CSF nucleic acid amplification testing(NAAT),criteria used for testing and turnaround times including the impact of the coronavirus disease 2019 pandemic.RESULTS Of 196/212(92%)microbiology laboratories responded;63/196(32%)were excluded from final analysis as they had no on-site microbiology laboratory and outsourced their samples.Of 133 Laboratories included in the study,47/133(35%)had onsite facilities for viral CSF NAAT.Hospitals currently undertaking onsite NAAT(n=47)had much faster turnaround times with 39 centers(83%)providing results in≤24 h as compared to those referring samples to neighboring laboratories(5/86;6%).CONCLUSION Onsite/near-patient rapid NAAT(including polymerase chain reaction)is recommended wherever possible to optimize patient management in the acute setting.
文摘目的基于血液筛查核酸检测反应性献血者的HBV感染的确认,探讨核酸检测反应性献血者的归队策略。方法联合应用自建的高灵敏度核酸检测体系、血液核酸筛查等多种核酸检测(NAT)方法,并结合血清学检测、献血者随访,对核酸检测反应性(NAT-yield)献血者中的HBV感染进行确认和感染状态识别。依据确认的HBV感染血浆样本,比较不同确认方法、确认指标或指标组合对HBV感染确认的效果。结果2010年11月—2021年2月,在血液筛查检出的876位NAT-yield献血者中共确认HBV感染者511人(OBI 451人,急性早期HBV感染者27人,不能确认感染者33人,无感染者30人,不能确认HBV感染者335人)。采用单检系统对混检系统检出的HBV感染血浆进行复测的检出率为96.6%,明显高于混检系统对单检系统检出的HBV DNA反应性(HBV DNA R)组和鉴别试验无反应性(NDR)组的复测检出率(76.4%和55.7%)(P<0.05)。NDR样本在模式2(ID×5+鉴别×2)下复测检出率(65.2%)高于模式1(ID×2+鉴别×1)(39.2%)(P<0.05);2种单检复测模式下的HBV DNA R样本复测检出率无明显差异(P>0.05),但均明显高于NDR样本(P<0.05)。回溯OBI献血者既往NAT数据,有46%经历多次NAT检测而未能检出。有59.1%OBI献血者随访检不出HBV DNA。OBI献血者中抗-HBc+占比为90.2%,单独抗-HBc+为49.2%,远高于不能确认感染组(P<0.05);HBeAg、抗-HBe和抗-HBc IgM在OBI和不能确认感染组中的比例极低且无差异(P>0.05)。结论近60%的NAT-yield献血者可以确认HBV感染。为保证献血者归队的安全性,需要更高灵敏度的HBV DNA确证技术提高HBV感染的确认率。抗-HBc是NAT-yield献血者OBI风险排查和归队评估最重要的血清学指标。
文摘目的通过危害分析及关键控制点(hazard analysis and critical control point,HACCP)体系系统化控制核酸检测实验室质量,以期能够实现核酸检测实验室质量的提升,为有效制定核酸检测质量控制对策提供借鉴。方法根据HACCP体系初步探究核酸检测实验室质量控制,重点分析HACCP体系在核酸标本接收、准备试剂、提取标本核酸、核酸扩增、结果判读、核酸检测结果报告环节制定的监控程序、验证措施以及纠偏措施等重要控制点。结果通过分析2021年12月-2022年12月的检测数据,结果显示,各批次核酸检测阳性质控品均满足判定规则,每一批次的实验均处于在控状态。对HACCP体系下实验室数据进行统计,计算其标准差、均值以及变异系数,对箱型图绘制后,展开离群值检验,结果发现,均值没有产生离群值。结论核酸检测实验室中应用HACCP体系,能够有效控制核酸检测实验室质量,全面掌控核酸检测实验室重要控制点,实现实验室生物安全水平与质量的提升,保证核酸检测实验室质控的有效性与科学性,继而有效控制核酸检测实验室质量。
