Balamurugan Vinayagamurthy, Jayappa Kallesh Danappa, Hosamani Madhusudhan, Bhanuprakash Veerakyathappa, Venkatesan Gnanavel, Singh Raj Kumar
Division of Virology, Indian Veterinary Research Institute, Campus Mukteswar-263138, Nainital District, Uttarakhand, India.
J Vet Diagn Invest. 2009 Mar;21(2):225-31. doi: 10.1177/104063870902100208.
Sheeppox and goatpox are economically important viral diseases of sheep and goats, respectively. Both diseases are reportable to the World Organization for Animal Health. To implement a control and eradication program for these diseases, a rapid and user-friendly diagnostic tool is imperative for screening. Therefore, in the present study, TaqMan quantitative polymerase chain reaction (qPCR) and conventional PCR assays targeting the DNA polymerase (DNA pol) gene were developed for the detection of Capripoxvirus DNA from clinical specimens of sheep and goats. The 2 assays used different primer sets. Conventional PCR yielded a specific product of 134 bp, whereas qPCR yielded a 180-bp product. The specificity of amplified DNA pol gene products was confirmed by their size and by sequence analysis. The 2 assays were specific for Sheeppox virus and Goatpox virus. However, in comparison to conventional PCR, the qPCR was more rapid, specific, and 100 times more sensitive, with a detection limit as low as 0.042 pg of purified DNA. The qPCR assay was more sensitive (84.05%) than conventional PCR (76.06%) when used on clinical samples (n = 71) from sheep and goats.
绵羊痘和山羊痘分别是绵羊和山羊的重要经济病毒性疾病。这两种疾病均须向世界动物卫生组织报告。为实施针对这些疾病的防控和根除计划，一种快速且用户友好的诊断工具对于筛查至关重要。因此，在本研究中，开发了针对DNA聚合酶（DNA pol）基因的TaqMan定量聚合酶链反应（qPCR）和常规PCR检测方法，用于从绵羊和山羊的临床样本中检测山羊痘病毒DNA。这两种检测方法使用了不同的引物组。常规PCR产生了一条134 bp的特异性产物，而qPCR产生了一条180 bp的产物。通过产物大小和序列分析证实了扩增的DNA pol基因产物的特异性。这两种检测方法对绵羊痘病毒和山羊痘病毒具有特异性。然而，与常规PCR相比，qPCR更快、更特异，灵敏度高100倍，检测限低至0.042 pg纯化DNA。当用于绵羊和山羊的临床样本（n = 71）时，qPCR检测方法比常规PCR（76.06%）更灵敏（84.05%）。
