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犬恶丝虫抗体dot-ELISA检测方法的建立
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刘德昊1，杨光友1*，王成东2，杨智2，罗娌2，刘礼2，兰镜超2，黄祥明2
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（1．四川农业大学 动物医学院，四川 雅安625014;
2．成都大熊猫繁育研究基地，四川 成都610081）
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摘要：
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为快速诊断犬恶丝虫病，建立了犬恶丝虫抗体dot-ELISA检测方法，并与阻断试验、交叉反应试验、重复性试验、琼脂扩散试验和平行对照试验进行了比较。结果显示，制备的犬恶丝虫虫体粗制抗原与犬钩虫、犬弓首蛔虫和犬蠕形螨感染犬的阳性血清无交叉反应，检出犬恶丝虫阳性血清的最高抗体效价为1∶1024，重复性好；建立的犬恶丝虫病dot-ELISA诊断方法的灵敏度比琼脂扩散试验和美国IDEXX实验室制备的犬恶丝虫病诊断试剂盒分别高2048倍和48倍。 |
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关键词：
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犬恶丝虫；粗制抗原；斑点酶联免疫吸附试验；抗体 |
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中图分类号：S 852.731 文献标识码：
A 文章编号：1673-4696(2008)02-0106-05 |
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Establishment of a dot-ELISA for detection of antibody
against Dirofilaria immitis |
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LIU De-hao1，YANG Guang-you1，WANG
Cheng-dong2，YANG Zhi2，
LUO Li2，LIU Li2，LAN Jing-chao2，HUANG
Xiang-ming2 |
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(1.College of Veterinary Medicine，Sichuan Agricultural University,Ya’an 625014,China；
2.Chengdu Research Base for Giant Panda Breeding,Chengdu 610081,China) |
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Abstract: In order to rapidly diagnose dirofilariasis in dog,a dot-ELISA for detection of antibody against Dirofilaria immitis was established using raw antigen extracted from D.immitis and evaluated by interruption test，cross-reaction test，reproducibility test,agar diffusion test and parallel control test.There was no cross-reaction of the prepared raw antigen with the positive sera from dogs infected respectively with Ancylostoma caninum，Toxocara canis and Demodex canis and the highest titer of the positive serum from dogs infected with D.immitis was 1∶1024 detected by the dot-ELISA,which had good reproducibi-lity.The sensitivity of the dot-ELISA was 2048 times and 48 times higher than that of the agar diffusion test and Canine Heartworm Antigen Test Kit made by American IDEXX Laboratory,respectively. |
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Key words: Dirofilaria immitis；raw antigen；dot-ELISA；antibody |
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