New approaches for the molecular characterization of canine parvovirus type 2 strains
New approaches for the molecular characterization of canine parvovirus type 2 strains
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DOI：
10.1111/j.1439-0450.2005.00869.x
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发表时间：
2005-09-01
期刊：
影响因子：
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通讯作者：
Buonavoglia, C
中科院分区：
文献类型：
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作者：
Decaro, N;Elia, G;Buonavoglia, C
犬细小病毒2型（CPV-2）的特征有时是模糊的，通常需要一种以上的技术来确定性预测病毒类型。考虑到2a型和2b型之间以及2b型和Glu-426突变体（2c型）之间在VP 2蛋白基因中遇到的单核苷酸多态性，开发了两种不同的小沟结合物（MGB）探针检测法用于快速鉴定CPV-2变体。通过能够检测所有CPV-2型的实时聚合酶链反应（PCR）检测，对从腹泻犬中采集的共计315份样本进行了CPV-2筛查。为比较CPV-2型特异性检测方法与传统方法（单克隆抗体血凝抑制法、PCR-限制性片段长度多态性（RFLP）法、序列分析法）对CPV-2型抗原特异性的预测效果，对203份CPV-2型阳性标本进行了分析。结果显示，MGB探针检测与常规方法的结合之间的一致性为100%，其中116例被鉴定为2a型，32例为2b型，55例为2c型。因此，相对于目前使用的耗时更长的检测方法，MGB探针检测是一种快速、可靠的CPV-2抗原特异性预测工具。
Characterization of the canine parvovirus type 2 (CPV-2) is sometimes ambiguous, frequently requiring more than one technique for definitive prediction of the viral type. Taking into account the single-nucleotide polymorphisms encountered in the VP2-protein gene between types 2a and 2b and between type 2b and Glu-426 mutant (type 2c), two different minor groove binder (MGB) probe assays were developed for rapid identification of the CPV-2 variants. A total of 315 samples collected from dogs with diarrhoea were screened for CPV-2 by a real-time polymerase chain reaction (PCR) assay capable of detecting all CPV-2 types. In order to compare the type-specific assays with the traditional techniques [haemagglutination inhibition with monoclonal antibodies, PCR-restriction fragment-length polymorphism (RFLP), sequence analysis] for prediction of CPV-2 antigen specificity, the 203 samples tested CPV-2 positive were analysed using the different methods. The results showed a 100% concordance between the MGB probe assays and the combined conventional methods, with 116 samples characterized as type 2a, 32 as type 2b and 55 as type 2c. Therefore, the MGB probe assays represent a quick, reliable tool for prediction of CPV-2 antigen specificity, with regard to the more time-consuming assays currently used.