公告:
三種套組檢測豬隻口蹄疫非結構性蛋白抗體之比較
詳細內容:
作者:豬瘟研究組 李 璠 助理研究員
本實驗係對三種檢測豬隻口蹄疫非結構性蛋白抗體檢測套組進行比較。 第一種套組是以大腸桿菌表現的3ABC為抗原的CHEKIT FMD-3ABC套組, 第二種是以合成3B為抗原的UBI FMDV NS EIA套組, 第三種是以桿狀病毒表現的3AB為抗原的DVIVR NSP ELISA套組。 實驗中將9頭免疫與2頭未免疫豬隻接種口蹄疫O/Taiwain/97病毒株,動物皆出現水泡病變。動物每週採血,連續採集血清13週作為檢測樣本。 檢測樣本亦包含302支未免疫與102支重複免疫豬隻血清,以評估套組之特異性。 結果顯示CHEKIT、UBI、DVIVR三種套組的特異性分別為99.3%、96.8%及96.8%。DVIVR套組的分析敏感性(Analytical sensitivity)為三者之最。 CHEKIT/UBI、CHEKIT/DVIVR、UBI/ DVIVR之間檢測結果的一致性,經kappa statistics計算的數值分別為0.58、0.46、0.67。某些豬隻測得的非結構性蛋白抗體反應呈現短暫或間歇性出現的情形。 由於測得的豬隻免疫反應並未完全反映出動物的感染狀況,當應用這些檢測方式汰除豬群中的感染動物時,可能會受到某些限制。    Comparison of three enzyme-linked immunosorbent assays (ELISA) kits on their efficacy to detect non-structural protein (NSP) antibodies in naive, foot-and-mouth disease virus infected and vaccinated porcine sera was conducted. The CHEKIT FMD-3ABC is based on E. coli-expressed 3ABC antigen, the UBI FMDV NS EIA is based on synthetic 3B peptide, and the DVIVR NSP ELISA is based on baculovirus-expressed 3AB antigen. Nine-vaccinated and two non-vaccinated six-month-old pigs were challenged with foot-and-mouth disease virus O/Taiwan.97 strain. They developed vesicular lesions and their sera were collected weekly until thirteen weeks post challenge. Sera from 302 naive and 102 repeatedly vaccinated healthy pigs were also examined with the kits for evaluation of specificity. The results revealed that the specificity of the CHEKIT kit, the UBI kit and the DVIVR kit was 99.3%, 96.8% and 96.8%, respectively. The DVIVR kit had the highest analytical sensitivity. The kappa statistics, 0.58, 0.46 and 0.67 on detection of all 623 sera of the present study showed moderate agreement between CHEKIT/UBI, CHEKIT/DVIVR and UBI/ DVIVR, respectively. Measurable NSP-specific antibodies in some of the infected vaccinated pigs were sustained either for shorter periods or in intermittent patterns. Immune responses to NSPs in pigs did not completely reflect previous exposure of animals and therefore the detection systems of antibodies to NSPs might have their limitation for stamping exposed pigs out of the population.
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