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目的评价检测内脏利什曼病特异抗体的胶体金免疫层析试条检测我国动物源型内脏利什曼病流行区犬血样的效果。方法以杜氏利什曼原虫前鞭毛体粗抗原作为包被抗原,以链球菌G蛋白标记胶体金制成检测特异抗体的免疫层析试条(immunochromatographic strip test,ICT)。用该试条检测甘肃省陇南市武都区(动物源型黑热病流行区)和甘肃省漳县(非流行区)犬的腿弯处静脉血样品,同时用镜检法、PCR和酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)分别检测犬骨髓、静脉血和血清样本,并比较各种检测方法的敏感性差异。结果对动物源型内脏利什曼病流行区的105头犬的骨髓涂片镜检,阳性检出率为7.62%(8/105)。应用PCR法的阳性检出率为48.57%(51/105);检测非流行区30份犬静脉血,1份为阳性,阳性检出率为3.33%(1/30)。试条法和ELISA法检测内脏利什曼病流行区的105份犬血样的阳性检出率分别为32.38%(34/105)和33.33%(35/105);检测内脏利什曼病非流行区的30份犬血样均为阴性。镜检法的阳性检出率低于PCR法、试条法和ELISA法(x~2=43.58、20.12、21.32,P<0.05),PCR法的阳性检出率高于镜检法、试条法和ELISA法(x~2=43.58、5.71、5.04,P<0.05),试条法和ELISA法阳性检出率无统计学差异(x~2=0.02,P>0.05)。结论快速检测内脏利什曼病的胶体金免疫层析试条适用于检测动物源型内脏利什曼病流行区犬血样中特异性抗体。
Objective To evaluate the effect of colloidal gold immunochromatographic test strip for the detection of visceral leishmaniasis-specific antibodies in the detection of canine blood samples from animal-derived visceral leishmaniasis in China. Methods Immunochromatographic strip test (ICT) was performed to detect the specific antibody using Leishmania donovani promastigote crude antigen as coating antigen and Streptococcus G protein labeled colloidal gold. The test strips were used to detect the venous blood of the dog in Wudu District of Longnan City, Gansu Province (epidemic area of animal-derived kala-azar) and Zhangxian (non-endemic area) of Gansu Province. At the same time, The enzyme-linked immunosorbent assay (ELISA) was used to detect canine bone marrow, venous blood and serum samples respectively, and to compare the sensitivities of different detection methods. Results The positive detection rate of bone marrow smear of 105 dogs in endemic visceral leishmaniasis area was 7.62% (8/105). The positive rate of PCR was 48.57% (51/105). In 30 non-endemic areas, 30 samples of canine venous blood were positive. The positive rate was 3.33% (1/30). The positive detection rates of 105 samples of canine blood from the prevalence area of visceral leishmaniasis were 32.38% (34/105) and 33.33% (35/105) respectively by the test strip method and the ELISA method. The detection of visceral leishmaniasis was non-epidemic District 30 dog blood samples were negative. The positive detection rate of microscopic examination was lower than that of PCR method, strip method and ELISA method (x ~ 2 = 43.58,20.12,21.32, P <0.05). The positive rate of PCR method was higher than that of microscopy, (X ~ 2 = 43.58,5.71,5.04, P <0.05). There was no significant difference between the two methods (x ~ 2 = 0.02, P> 0.05). Conclusion Colloidal gold immunochromatographic strips for the rapid detection of visceral leishmaniasis are suitable for the detection of specific antibodies in canine blood samples from endemic visceral leishmaniasis.