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首页> 外文期刊>Journal of Virological Methods >Development of one-step multiplex RT-PCR method for simultaneous detection and differentiation of foot-and-mouth disease virus serotypes O, A, and Asia 1 circulating in Vietnam.
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Development of one-step multiplex RT-PCR method for simultaneous detection and differentiation of foot-and-mouth disease virus serotypes O, A, and Asia 1 circulating in Vietnam.

机译:一步多路RT-PCR方法的开发,用于同时检测和区分在越南传播的O,A和Asia 1型口蹄疫病毒。

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A one-step multiplex RT-PCR method using new primers was developed for the simultaneous detection and differentiation of Vietnamese FMDV serotypes O, A, and Asia 1 directly from clinical samples. The RT-PCR method used a cocktail of one universal minus-sense primer designed in the 2B gene and three serotype-specific plus-sense primers designed in the hypervariable regions of the capsid VP1 coding gene of FMDV. These serotype-specific primer pairs amplified 658, 535, and 427 bp PCR products corresponding to FMDV serotypes O, Asia 1, and A, respectively. In this study, six well-characterized FMDV strains belonging to serotypes O, A, and Asia 1 were used as reference strains for validation tests. Among these six FMDV strains were three vaccine strains for type O (O1/Manisa), A (A22/Iraq), and Asia 1 (As1/Shamir/89). The other reference strains included one pandemic strain of FMDV serotype Asia 1 (Asia1/MOG/05) and two pandemic strains of FMDV serotype O (O/UKG/34/2001 and O/SKR/2000). For field application, 37 positive-clinical samples and 18 cell culture-adapted viruses belonging to serotypes O, A, and Asia 1, as confirmed previously by antigen ELISA for FMDV detection, were used. The present method showed high sensitivity and specificity and can be adapted for detection and typing of FMDV serotypes O, A, and Asia 1 circulating in Vietnam.Digital Object Identifier http://dx.doi.org/10.1016/j.jviromet.2011.04.027
机译:开发了一种使用新引物的一步多路RT-PCR方法,用于直接从临床样品中同时检测和区分越南FMDV血清型O,A和Asia 1。 RT-PCR方法使用在2B基因中设计的一种通用负义引物和在FMDV衣壳VP1编码基因的高变区中设计的三种血清型特异性正义引物的混合物。这些血清型特异性引物对分别扩增了658、535和427 bp的FMDV血清型O,亚洲1和A的PCR产物。在这项研究中,使用了六个O型,A型和Asia 1型的特征明确的FMDV菌株作为验证试验的参考菌株。在这六种FMDV菌株中,有三种分别针对O型(O1 / Manisa),A型(A22 /伊拉克)和亚洲1型(As1 / Shamir / 89)的疫苗株。其他参考菌株包括一种FMDV血清型亚洲1的大流行毒株(Asia1 / MOG / 05)和两种FMDV血清型O的大流行毒株(O / UKG / 34/2001和O / SKR / 2000)。对于现场应用,使用了37种阳性临床样品和18种适应血清培养的病毒,分别属于O,A和Asia 1型,已通过抗原ELISA确认用于FMDV检测。本方法显示出高灵敏度和特异性,可适用于在越南流通的FMDV血清型O,A和Asia 1的检测和分型。数字对象标识符http://dx.doi.org/10.1016/j.jviromet.2011.04 .027

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