犬恶丝虫感染胶体金试纸条检测方法的建立及应用

Devepoment of colloidal gold strip for detection of Dirofilaria immitis in dogs

目的建立犬恶丝虫血清抗体检测胶体金试纸条方法。方法利用胶体金标记的金黄色葡萄球菌A蛋白(SPA)作为金标抗体,犬恶丝虫MTFP重组蛋白作为诊断抗原标记检测线(T线),兔抗SPA抗体标记质检线(C线),对建立的各项条件进行优化,制备胶体金试纸条;对建立的方法进行特异性、敏感性、稳定性验证试验。用建立的胶体金试纸条检测方法对62份犬血清样本进行检测。结果所制备的胶体金试纸条敏感性为1∶8,重复性良好,检测犬等孢球虫阳性血清、犬蛔虫阳性血清结果均为阴性。应用制备的胶体金试纸条检测62份犬血清,阳性样本4份,阴性样本58份,阳性检出率6.45%。结论建立的犬恶丝虫感染胶体金试纸条检测方法具有特异性、稳定性等特点,为犬恶丝虫病的诊断提供了一种新的检测方法。

Objective To develop a new method of colloidal gold strip for the detection of Dirofilaria immitis in dogs. Methods Staphylococcal protein A（SPA）was labeled by colloidal gold as the antibody-colloidal.The test line（T line）was labeled by the MTFP antigen and control line（C line）was labeled by rabbit anti-SPA antibody.The colloidal gold strip was constructed by optimizing test conditions.The specificity,sensitivity and stability of colloidal gold strip were performed.And 62 serum samples from dogs were tested by the established method. Results The sensitivity of the established colloidal gold strip was 1∶8.There were no cross reaction with Isospora canis and Toxocara canis.Out of 62 serum samples tested,4 were antibody positive for Dirofilaria immitis with positive rate of 6.4%. Conclusions The colloidal gold strip method to detect Dirofilaria immitis infection in dogs has the advantages of high specificity,sensitivity and stability.It provides a new method for detection Dirofilaria immitis infection in dogs.