Institute of Animal Sciences and Veterinary Medicine and Shanghai Key Laboratory of Agricultural Genetics and Breeding, Shanghai Academy of Agricultural Sciences, SAAS, Shanghai, China.
Intervirology. 2013;56(1):55-9. doi: 10.1159/000341601. Epub 2012 Oct 5.
To develop an effective diagnostic kit, based on a competitive ELISA-based system (cELISA), for detecting serum antibody against peste des petits ruminants virus (PPRV).
Epitope peptides of the nucleocapsid (N) protein of Tibetan PPRV were synthesized chemically and injected into rabbits to prepare hyperimmune antisera. Test sera were incubated simultaneously with hyperimmune antisera and added to the wells of ELISA plates coated previously with recombinant N protein. Horseradish peroxidase-conjugated goat anti-rabbit antibody was employed to detect the quantity of hyperimmune antisera combined with recombinant N protein.
A cELISA has been developed for monitoring PPRV infections with a cutoff value of 35. Relative sensitivity and specificity values of the epitope-based cELISA were 96.18 and 91.29%, respectively, when compared with a commercial cELISA kit in a test involving 1,039 serum samples.
We report an efficient method for preparing antibody suitable for incorporation into a cELISA that can be used routinely for the detection of PPRV antibodies in serum samples. The method eliminated the requirement for virus culture and monoclonal antibody preparation, reduced the biorisk posed by virus-dependent manipulations, and the performance of the resultant cELISA compared favorably with a commercially available cELISA kit.
基于竞争 ELISA 系统（cELISA），开发一种用于检测小反刍兽疫病毒（PPRV）血清抗体的有效诊断试剂盒。
化学合成小反刍兽疫病毒西藏株核衣壳（N）蛋白的表位肽，并注射到兔子体内制备高免血清。将待检血清与高免血清同时孵育，然后加入到已包被重组 N 蛋白的 ELISA 板孔中。辣根过氧化物酶标记的山羊抗兔抗体用于检测与重组 N 蛋白结合的高免血清量。
建立了一种用于监测小反刍兽疫病毒感染的 cELISA，其截断值为 35。与商业 cELISA 试剂盒相比，在涉及 1039 份血清样本的试验中，基于表位的 cELISA 的相对灵敏度和特异性值分别为 96.18%和 91.29%。
我们报告了一种制备适合 cELISA 检测的抗体的有效方法，可常规用于检测血清样本中的小反刍兽疫病毒抗体。该方法消除了病毒培养和单克隆抗体制备的需求，降低了病毒依赖性操作带来的生物风险，并且所得 cELISA 的性能与市售 cELISA 试剂盒相当。
