猪轮状病毒免疫荧光制剂质量检测与评价

The Quality Assay and Evaluation on ImmunofluorescentAntibody of Porcine Rotavirus

对猪轮状病毒（ＰＲＶ）Ｂｏｈｌ株纯化抗原制备的荧光抗体（ＦＡ）和标准阳、阴性片做了较全面的检验。ＦＡ的Ｆ／Ｐ比值在１．５～１．９８恰当范围。染色效价高达１：２５６。使用４或８个单位时，既能消除非特异染色，又有强烈特异染色效果。ＦＡ抑制试验、吸收试验、类症鉴别试验证明有高度特异性。ＦＡ敏感性试验表明，可以在接种后５ｈ检出细胞培养物中的ＰＲＶ抗原，比ＣＰＥ出现早１０ｈ以上。稳定性和保存期试验表明，冻干ＦＡ或湿剂型ＦＡ２年内效价降低不明显。于２０～２８℃运输７ｄ，又４℃存放３８ｄ，按规程（草案）规定稀释仍有良好的染色性能。应用ＦＡ检查ＰＲＶ人工感染猪２２头、已知混合感染猪２１头，检出率１００％（４３／４３）。猪传染性胃肠炎（ＴＧＥ）１９例、猪流行性腹泻（ＰＥＤ）１１例及猪大肠杆菌病猪１０例全部为ＰＲＶ阴性。对１００７份腹泻病例进行检查，ＰＲＶ阳性率平均为１９．６６％。对阳、阴性参考标准片用国外引进的制品检查认为有良好的特异性并符合标准。与美国ＦＡ比较试验表明，虽两者阳性、阴性结果相一致，但自制ＦＡ着染强度更佳。

The FA prepared with purified antigen of porcine rotavirus Bohl strainand standard positive and negative smears were assayed.F/P ratio of FA ranged proper-ly from 1.5 to 1.8, and its staining titer was 1 :256.FA could not only eliminate thenonspecific staining but also had strong specific staining effect when 4 or 8 units wereused. FA inhibitory test, absorption test and analogous symptomatic differential testshowed that FA had a high specificity. FA sensitivity test indicated that PRV antigen incell cultures could be detected in a short time (5 h) after virus inoculation, which wasover 10 h earlier than the appearence of CPE. The titers of lyophilized FA or wet type ofFA did not decrease obviously within 2 years, which had been proved by stability andstorage test.22 pigs infected artificially with PRV and 21 pigs knowm with mixed infec-tion were detected with FA,the detectable rate was 100%(43/43).19 cases of TGE,11 of PED and 10 of porcine colibacillosis were all PRV negative。 In 1007 pigs sufferedfrom diarrhea the average positive rates of PRV was l9.66%.