| 摘要: |
| 【目的】建立快速、准确检测鸭副粘病毒病的方法。【方法】根据GenBank中发表的新城疫F基因序列设计1对引物，扩增鸭副粘病毒融合蛋白基因727 bp的特异性片段，进行特异性试验和敏感性试验，建立了鸭副粘病毒病的RT-PCR诊断方法。应用建立的诊断方法，对从山东不同地区分离的20株疑似鸭副粘病毒毒株和150份疑似感染副粘病毒鸭的病变组织进行了检测。【结果】特异性试验表明，建立的RT-PCR检测方法能够从鸭副粘病毒SDFCH株中扩增到727 bp的特异性片段，而对鸭瘟病毒、H9N2亚型禽流感病毒、鸭肝炎病毒的扩增结果均为阴性；敏感性试验表明，该法最低检出量的cDNA质量浓度为3 pg/μL；山东不同地区20株疑似鸭副粘病毒分离株中有15份为阳性；150份疑似感染副粘病毒鸭的病变组织有125例为阳性。【结论】建立的鸭副粘病毒病的RT-PCR诊断方法，且有快速、准确、特异性强、敏感性高的特点。 |
| 关键词: 鸭 副粘病毒 RT-PCR |
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| 基金项目:山东省科技攻关项目（2007GG30009001） |
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| Establishment and application of RT-PCR diagnostic method for duck paramyxovirus disease |
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| Abstract: |
| 【Objective】A rapid and accurate detection method was established for duck paramyxovirus disease.【Method】The fusion (F) gene fragment (727 bp) of duck paramyxovirus was amplified by RT-PCR diagnostic method using a pair of primers,followed by specificity test and sensitivity test.Then 20 suspected strains of duck paramyxovirus and 150 samples of infectious or diseased tissues of ducks paramyxovirus were detected by the RT-PCR method.【Result】 The specificity test showed that the method could specially amplify the 727 bp gene specific fragment of duck paramyxovirus SDFCH strain,while the amplification results of duck plague virus,H9N2 subtype avian influenza virus,duck hepatitis virus were negative.Sensitivity test showed that the minimum detectable amount of cDNA concentration was 3 pg/μL.15 from the 20 isolated strains were paramyxovirus (positive) while 125 from the 150 tissue samples were positive.【Conclusion】 The established RT-PCR diagnostic method appeared to be fast,accurate,highly sensitive and specific for the purpose. |
| Key words: duck paramyxovirus RT-PCR |
