摘要
以５Ｂ８株抗ＤＨＢｓＡｇ特异性单克隆抗体作为包被抗体和酶标抗体，首次建立了检测血清中ＤＨＢｓＡｇ的单抗夹心ＥＬＩＳＡ．对纯化ＤＨＢｓＡｇ的最小检出量为１０ｎｇ／ｍｌ．对雏鸭肝炎病毒、鸭瘟病毒等９种禽类常见的病毒病抗原及人乙肝病毒表面抗原（ＨＢｓＡｇ）均不发生反应，且能被多克隆的兔抗ＤＨＢｓＩｇＧ及５Ｂ８株单抗本身所阻断。应用本试验方法调查江苏省部分地区麻鸭ＤＨＢＶ的感染率分别为：高邮２４．５％、常州２１．７％、仪征４０．３％、启东５０．０％。１日龄雏鸭静脉接种ＤＨＢＶ后，经本法检测，结果ＤＨＢｓＡｇ阳性率从４０％上升到７９％，６个月后又下降至６９％。
Abstract A monoclonal antibody Based sandwich ELISA was developed for detection of DHBsAg in duck sera. McAb 5B8 with specificity to DHBsAg was used as coating antibody and preparation of the HRPO conjugated antibody. The minimum limit of purified DHBsAg detected by this method was 10ng vital surface protein/ml. HBsAg and the pathogens of nine kinds of avain virus disease were all negative tested by this method. Besides,this assay was inhibited by McAb 5B8 itself and rabbit anti-DHBs IgG.4 groups of duck sera from different area in Jiangsu Province had been detected for DHBsAg by ELISA. The positive rates were 24. 5% (24/98) in Gaoyou,21.7×(13/60) in Changzhou,40.3% (22/67) in Yizheng, 50. 0% (9/18) in Qidong, the results showed that when the ducks infectted with DHBV were inoculated intravenously with DHBV at 1 day of age, DHBV-infected rate increasted from 40% to 79%, but decreased to 69% after 6 months.
基金
国家自然科学基金
