Chambers M A, Swango L J, Wright J C
Department of Small Animal Internal Medicine, School of Veterinary Medicine, Tuskegee University, AL 36088, USA.
J Vet Diagn Invest. 1996 Apr;8(2):196-201. doi: 10.1177/104063879600800209.
An indirect fluorescent antibody (IFA) test was developed using a novel format of Borrelia burgdorferi organisms adhered to a monolayer of cultured endothelial cells derived from an equine tumor. Sensitivity and specificity of the new IFA test for detecting anti-B, burgdorferi antibodies were evaluated using sera from dogs inoculated with live B. burgdorferi or vaccinated with B. burgdorferi bacterin or leptobacterins and from unvaccinated specific-pathogen-free (SPF) dogs. To compare the new IFA test with existing tests, serum samples were submitted to independent laboratories to be tested by enzyme-linked immunosorbent assay (ELISA) and a traditional IFA test. Samples were also tested with 2 commercially available membrane-bound ELISA kits. Both Borrelia-inoculated dogs and dogs vaccinated with B. burgdorferi bacterin developed levels of antibody detectable by the new IFA test. Dogs vaccinated with a combination canine vaccine or leptobacterin for food animal use developed detectable levels of antibody against Leptospira but remained seronegative for Borrelia by the new IFA test, as did the unvaccinated SPF dogs. The new IFA test was sensitive, detecting antibodies against B. burgdorferi as early as 7 days postinoculation. It was also specific, showing no cross-reactivity with anti-Leptospira antibodies induced by vaccination with leptobacterins. The new IFA test compared favorably with both the standardized traditional IFA test and ELISA. Results from both membrane-bound ELISA kits were not consistent when compared with each other or with the new IFA test. The new IFA test had low nonspecific fluorescence, which made it easier to evaluate and reduced the human error and variability of test results.
利用一种新的格式开发了一种间接荧光抗体（IFA）检测方法，该方法使用粘附于源自马肿瘤的培养内皮细胞单层的新型伯氏疏螺旋体生物体。使用接种活伯氏疏螺旋体、接种伯氏疏螺旋体菌苗或钩端螺旋体菌苗的犬以及未接种的无特定病原体（SPF）犬的血清，评估这种新型IFA检测方法检测抗伯氏疏螺旋体抗体的敏感性和特异性。为了将新型IFA检测方法与现有检测方法进行比较，将血清样本提交给独立实验室，通过酶联免疫吸附测定（ELISA）和传统IFA检测方法进行检测。样本还使用两种市售的膜结合ELISA试剂盒进行检测。接种伯氏疏螺旋体的犬和接种伯氏疏螺旋体菌苗的犬均产生了可被新型IFA检测方法检测到的抗体水平。接种用于食用动物的犬用联合疫苗或钩端螺旋体菌苗的犬产生了可检测到的抗钩端螺旋体抗体水平，但通过新型IFA检测方法对伯氏疏螺旋体仍呈血清阴性，未接种的SPF犬也是如此。新型IFA检测方法很敏感，在接种后7天就可检测到抗伯氏疏螺旋体抗体。它也具有特异性，与接种钩端螺旋体菌苗诱导的抗钩端螺旋体抗体无交叉反应。新型IFA检测方法与标准化的传统IFA检测方法和ELISA相比都具有优势。两种膜结合ELISA试剂盒的结果相互比较或与新型IFA检测方法比较时不一致。新型IFA检测方法的非特异性荧光较低，这使得评估更容易，并减少了人为误差和检测结果的变异性。
