用淋巴细胞杂交瘤技术,研制出的抗猪细小病毒单克隆抗体,采用酶联免疫吸附试验(ELISA)方法,对本地区所采得的308份猪血清进行检测,阳性检出率达66.6%。其中20～35日龄仔猪阳性检出率在80%以上;45日龄仔猪阳性检出率为75.5%;屠宰猪阳性...用淋巴细胞杂交瘤技术,研制出的抗猪细小病毒单克隆抗体,采用酶联免疫吸附试验(ELISA)方法,对本地区所采得的308份猪血清进行检测,阳性检出率达66.6%。其中20～35日龄仔猪阳性检出率在80%以上;45日龄仔猪阳性检出率为75.5%;屠宰猪阳性检出率55.2%。选择70份被检血清用已建立的检测猪细小病毒抗体的 ELISA 试验与传统的 HI 试验进行比较,其敏感性较高,ELISA 检出阳性率为85.7%,HI 检出阳性率为80%,前者较 HI 试验高出5.7个百分点。展开更多
用两株抗兔出血症病毒(RHDV)的单克隆抗体(McAb)混合包板,再以其中一株进行辣根过氧化物酶(HRP)标记,建立了检测 RHDV 的单抗夹心 ELISA 法。用本法和血凝(HA)试验检测51只实验感染致死兔的背肌、腿肌、淋巴结、脊髓和骨髓等5种组织中的...用两株抗兔出血症病毒(RHDV)的单克隆抗体(McAb)混合包板,再以其中一株进行辣根过氧化物酶(HRP)标记,建立了检测 RHDV 的单抗夹心 ELISA 法。用本法和血凝(HA)试验检测51只实验感染致死兔的背肌、腿肌、淋巴结、脊髓和骨髓等5种组织中的 RHDV。结果以骨髓中 RHDV 含量最高,且较稳定,OD 值集中在1.2～1.6范围内;淋巴结次之,但 OD 值分散;肌肉和脊髓的 OD 值和检出率均较低。ELISA 的检出率显著高于HA,二者符合率很低,尤其是脊髓和淋巴结。有些样品的 OD 值很高,而 HA 却为阴性。骨髓是胴体检测 RHDV的最佳材料。经测定,本法的工作浓度为1∶20,阳性判定界值(OD+3SD)为0.11。本法的样品处理简单,毋须浓缩,从样品处理到得出结果只需4.5h,与 HA 相比,更为灵敏和稳定。展开更多
Vitamin B 12 is an organometallic compound with important metabolic derivatives that act as cofactors of certain enzymes,which have been grouped into three subfamilies depending on their cofactors.Among them,methylmal...Vitamin B 12 is an organometallic compound with important metabolic derivatives that act as cofactors of certain enzymes,which have been grouped into three subfamilies depending on their cofactors.Among them,methylmalonyl-CoA mutase (MCM) has been extensively studied.This enzyme catalyzes the reversible isomerization of L-methylmalonyl-CoA to succinyl-CoA using adenosylcobalamin (AdoCbl) as a cofactor participating in the generation of radicals that allow isomerization of the substrate.The crystal structure of MCM determined in Propionibacterium freudenreichii var.shermanii has helped to elucidate the role of this cofactor AdoCbl in the reaction to specify the mechanism by which radicals are generated from the coenzyme and to clarify the interactions between the enzyme,coenzyme,and substrate.The existence of human methylmalonic acidemia (MMA) due to the presence of mutations in MCM shows the importance of its role in metabolism.The recent crystallization of the human MCM has shown that despite being similar to the bacterial protein,there are significant differences in the structural organization of the two proteins.Recent studies have identified the involvement of an accessory protein called MMAA,which interacts with MCM to prevent MCM's inactivation or acts as a chaperone to promote regeneration of inactivated enzyme.The interdisciplinary studies using this protein as a model in different organisms have helped to elucidate the mechanism of action of this isomerase,the impact of mutations at a functional level and their repercussion in the development and progression of MMA in humans.It is still necessary to study the mechanisms involved in more detail using new methods.展开更多
文摘用淋巴细胞杂交瘤技术,研制出的抗猪细小病毒单克隆抗体,采用酶联免疫吸附试验(ELISA)方法,对本地区所采得的308份猪血清进行检测,阳性检出率达66.6%。其中20～35日龄仔猪阳性检出率在80%以上;45日龄仔猪阳性检出率为75.5%;屠宰猪阳性检出率55.2%。选择70份被检血清用已建立的检测猪细小病毒抗体的 ELISA 试验与传统的 HI 试验进行比较,其敏感性较高,ELISA 检出阳性率为85.7%,HI 检出阳性率为80%,前者较 HI 试验高出5.7个百分点。
文摘用两株抗兔出血症病毒(RHDV)的单克隆抗体(McAb)混合包板,再以其中一株进行辣根过氧化物酶(HRP)标记,建立了检测 RHDV 的单抗夹心 ELISA 法。用本法和血凝(HA)试验检测51只实验感染致死兔的背肌、腿肌、淋巴结、脊髓和骨髓等5种组织中的 RHDV。结果以骨髓中 RHDV 含量最高,且较稳定,OD 值集中在1.2～1.6范围内;淋巴结次之,但 OD 值分散;肌肉和脊髓的 OD 值和检出率均较低。ELISA 的检出率显著高于HA,二者符合率很低,尤其是脊髓和淋巴结。有些样品的 OD 值很高,而 HA 却为阴性。骨髓是胴体检测 RHDV的最佳材料。经测定,本法的工作浓度为1∶20,阳性判定界值(OD+3SD)为0.11。本法的样品处理简单,毋须浓缩,从样品处理到得出结果只需4.5h,与 HA 相比,更为灵敏和稳定。
基金Project (No.IN208411) partially supported by the PAPIIT-UNAM of Mexico
文摘Vitamin B 12 is an organometallic compound with important metabolic derivatives that act as cofactors of certain enzymes,which have been grouped into three subfamilies depending on their cofactors.Among them,methylmalonyl-CoA mutase (MCM) has been extensively studied.This enzyme catalyzes the reversible isomerization of L-methylmalonyl-CoA to succinyl-CoA using adenosylcobalamin (AdoCbl) as a cofactor participating in the generation of radicals that allow isomerization of the substrate.The crystal structure of MCM determined in Propionibacterium freudenreichii var.shermanii has helped to elucidate the role of this cofactor AdoCbl in the reaction to specify the mechanism by which radicals are generated from the coenzyme and to clarify the interactions between the enzyme,coenzyme,and substrate.The existence of human methylmalonic acidemia (MMA) due to the presence of mutations in MCM shows the importance of its role in metabolism.The recent crystallization of the human MCM has shown that despite being similar to the bacterial protein,there are significant differences in the structural organization of the two proteins.Recent studies have identified the involvement of an accessory protein called MMAA,which interacts with MCM to prevent MCM's inactivation or acts as a chaperone to promote regeneration of inactivated enzyme.The interdisciplinary studies using this protein as a model in different organisms have helped to elucidate the mechanism of action of this isomerase,the impact of mutations at a functional level and their repercussion in the development and progression of MMA in humans.It is still necessary to study the mechanisms involved in more detail using new methods.
