Foot-and-mouth disease(FMD) is a highly contagious and economically devastating disease of livestock, primarily affecting cattle, buffalo and pigs. FMD virus serotypes O, A and Asia1 are prevalent in India and systema...Foot-and-mouth disease(FMD) is a highly contagious and economically devastating disease of livestock, primarily affecting cattle, buffalo and pigs. FMD virus serotypes O, A and Asia1 are prevalent in India and systematic efforts are on to control and eventually eradicate the disease from the country. FMD epidemiology is complex due to factors like co-circulation, extinction, emergence and re-emergence of genotypes/lineages within the three serotypes, animal movement, diverse farm practices and large number of susceptible livestock in the country. Systematic vaccination, prompt diagnosis, strict biosecurity measures, and regular monitoring of vaccinal immunity and surveillance of virus circulation are indispensible features for the effective implementation of the control measures. Availability of suitable companion diagnostic tests is very important in this endeavour. In this review, the diagnostic assays developed and validated in India and their contribution in FMD control programme is presented.展开更多
目的通过Meta分析评价多重荧光定量PCR(multiplex real time polymerase chain reaction,MRT—PCR)方法在呼吸道病毒中常见呼吸道合胞病毒(respiratory syncytial virus,RSV)和腺病毒(adenovirus,ADV)感染的诊断价值,为临床...目的通过Meta分析评价多重荧光定量PCR(multiplex real time polymerase chain reaction,MRT—PCR)方法在呼吸道病毒中常见呼吸道合胞病毒(respiratory syncytial virus,RSV)和腺病毒(adenovirus,ADV)感染的诊断价值,为临床应用提供参考。方法检索数据库包括PubMed、EMBASE、Cochrane、万方、CNKI等,检索2010年1月至2018年1月关于MRT—PCR方法检测呼吸道病毒感染的中英文文献,由2位研究者独立筛选文献、提取资料,并采用QUADAS-2工具评价纳人研究的偏倚风险后,采用Meta—disc1.4对数据进行统计分析。结果本研究纳入10篇文献共2528例样本。Meta分析结果显示,MRT—PCR检测RSV的Sen合并=0.87(95%CI:0.83—0.90)、Spe合并=0.98(95%CI:0.97—0.98)、AUC=1.00。MRT—PCR检测ADV的Sen合并=0.64(95%CI:0.56—0.71)、Spe合并=0.99(95%CI:0.98—0.99)、AUC=0.99、Q=0.96。Deeks漏斗图检验结果提示无明显的发表偏倚。结论MRT-PCR方法检测RSV和ADV的灵敏度有待提高,但检测综合能力较好,对临床上呼吸道病毒感染的早期诊断具有一定的应用价值。展开更多
目的建立多重实时定量PCR(MRQ—PCR)同时快速检测血中大肠杆菌与白念珠菌DNA的方法,以评估肠屏障损伤可能导致的移位微生物的种类和程度并提供针对性用药的建议。方法选择大肠杆菌β-右旋半乳糖苷酶基因作为检测大肠杆菌的靶基因设...目的建立多重实时定量PCR(MRQ—PCR)同时快速检测血中大肠杆菌与白念珠菌DNA的方法,以评估肠屏障损伤可能导致的移位微生物的种类和程度并提供针对性用药的建议。方法选择大肠杆菌β-右旋半乳糖苷酶基因作为检测大肠杆菌的靶基因设计引物和探针,选择白念珠菌ITS2基因设计引物和探针。采用QIAamp DNA Blood Mini Kit试剂盒提取大肠杆菌与白念珠菌基因组DNA;建立25μlTaqMan探针MRQ-PCR反应体系;对18份模拟血标本及10份外科发热患者临床标本进行MRQ—PCR检测。结果引物和探针特异性好,大肠杆菌与白念珠菌标准曲线相关系数分别在0.994—0.999和0.994~0.998,扩增效率分别为0.894~1.022和0.905~1.028。标准样品检测限分别为大肠杆菌13.9拷贝/μl和白念珠菌0.8cfu/μl,两种菌的检测灵敏度分别为100%和99.69%,特异度分别为100%和94.73%,标准品中大肠杆菌与白念珠菌特异基因平均回收率分别为(101.89±5.69)%和(103.74±4.64)%,两种菌基因检测的批内变异系数分别为(13.14±10.27)%和(19.18±8.54)%,批间变异系数分别为(14.35±9.34)%和(18.31±10.25)%。全血标本中大肠杆菌与白念珠菌特异基因的检出限分别为12455.2拷贝/ml和800.3cfu/ml,平均回收率分别为(111.60±11.06)%和(99.96±6.16)%,两种菌基因检测的批内变异系数分别为(11.02±5.65)%和(8.14±7.29)%,平均批间变异系数分别为(12.88±7.59)%和(18.62±9.14)%。结论多重实时定量PCR可以同时快速、灵敏、特异地定量检测人全血标本中大肠杆菌与白念珠菌基因,准确度高、重复性好、节省血标本用量及检测成本、总检测时间缩短,在临床全血标本的真菌与细菌快速鉴别检测、抗微生物药物的针对性应用及疗效评估、肠屏障�展开更多
文摘Foot-and-mouth disease(FMD) is a highly contagious and economically devastating disease of livestock, primarily affecting cattle, buffalo and pigs. FMD virus serotypes O, A and Asia1 are prevalent in India and systematic efforts are on to control and eventually eradicate the disease from the country. FMD epidemiology is complex due to factors like co-circulation, extinction, emergence and re-emergence of genotypes/lineages within the three serotypes, animal movement, diverse farm practices and large number of susceptible livestock in the country. Systematic vaccination, prompt diagnosis, strict biosecurity measures, and regular monitoring of vaccinal immunity and surveillance of virus circulation are indispensible features for the effective implementation of the control measures. Availability of suitable companion diagnostic tests is very important in this endeavour. In this review, the diagnostic assays developed and validated in India and their contribution in FMD control programme is presented.
文摘目的通过Meta分析评价多重荧光定量PCR(multiplex real time polymerase chain reaction,MRT—PCR)方法在呼吸道病毒中常见呼吸道合胞病毒(respiratory syncytial virus,RSV)和腺病毒(adenovirus,ADV)感染的诊断价值,为临床应用提供参考。方法检索数据库包括PubMed、EMBASE、Cochrane、万方、CNKI等,检索2010年1月至2018年1月关于MRT—PCR方法检测呼吸道病毒感染的中英文文献,由2位研究者独立筛选文献、提取资料,并采用QUADAS-2工具评价纳人研究的偏倚风险后,采用Meta—disc1.4对数据进行统计分析。结果本研究纳入10篇文献共2528例样本。Meta分析结果显示,MRT—PCR检测RSV的Sen合并=0.87(95%CI:0.83—0.90)、Spe合并=0.98(95%CI:0.97—0.98)、AUC=1.00。MRT—PCR检测ADV的Sen合并=0.64(95%CI:0.56—0.71)、Spe合并=0.99(95%CI:0.98—0.99)、AUC=0.99、Q=0.96。Deeks漏斗图检验结果提示无明显的发表偏倚。结论MRT-PCR方法检测RSV和ADV的灵敏度有待提高,但检测综合能力较好,对临床上呼吸道病毒感染的早期诊断具有一定的应用价值。
文摘目的建立多重实时定量PCR(MRQ—PCR)同时快速检测血中大肠杆菌与白念珠菌DNA的方法,以评估肠屏障损伤可能导致的移位微生物的种类和程度并提供针对性用药的建议。方法选择大肠杆菌β-右旋半乳糖苷酶基因作为检测大肠杆菌的靶基因设计引物和探针,选择白念珠菌ITS2基因设计引物和探针。采用QIAamp DNA Blood Mini Kit试剂盒提取大肠杆菌与白念珠菌基因组DNA;建立25μlTaqMan探针MRQ-PCR反应体系;对18份模拟血标本及10份外科发热患者临床标本进行MRQ—PCR检测。结果引物和探针特异性好,大肠杆菌与白念珠菌标准曲线相关系数分别在0.994—0.999和0.994~0.998,扩增效率分别为0.894~1.022和0.905~1.028。标准样品检测限分别为大肠杆菌13.9拷贝/μl和白念珠菌0.8cfu/μl,两种菌的检测灵敏度分别为100%和99.69%,特异度分别为100%和94.73%,标准品中大肠杆菌与白念珠菌特异基因平均回收率分别为(101.89±5.69)%和(103.74±4.64)%,两种菌基因检测的批内变异系数分别为(13.14±10.27)%和(19.18±8.54)%,批间变异系数分别为(14.35±9.34)%和(18.31±10.25)%。全血标本中大肠杆菌与白念珠菌特异基因的检出限分别为12455.2拷贝/ml和800.3cfu/ml,平均回收率分别为(111.60±11.06)%和(99.96±6.16)%,两种菌基因检测的批内变异系数分别为(11.02±5.65)%和(8.14±7.29)%,平均批间变异系数分别为(12.88±7.59)%和(18.62±9.14)%。结论多重实时定量PCR可以同时快速、灵敏、特异地定量检测人全血标本中大肠杆菌与白念珠菌基因,准确度高、重复性好、节省血标本用量及检测成本、总检测时间缩短,在临床全血标本的真菌与细菌快速鉴别检测、抗微生物药物的针对性应用及疗效评估、肠屏障�