A new nanoPCR molecular assay for detection of porcine bocavirus
A new nanoPCR molecular assay for detection of porcine bocavirus
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一种检测猪博卡病毒的新型 nanoPCR 分子检测方法
DOI：
10.1016/j.jviromet.2014.02.029
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发表时间：
2014-06-15
影响因子：
3.1
通讯作者：
Cui, Shangjin
中科院分区：
文献类型：
--
作者：
Wang, Xiaoling;Bai, Aiquan;Cui, Shangjin
纳米颗粒辅助聚合酶链反应（nanoPCR）是一种快速扩增DNA的新方法，已被用于病毒检测。为检测猪博卡病毒（PBoV），根据GenBank中的NS 1基因序列设计了一对引物，建立了一种灵敏、特异的nanoPCR检测方法。在PBoV nanoPCR测定的优化条件下，nanoPCR测定比常规PCR测定灵敏100倍。nanoPCR测定的检测下限为约6.70 Chi 10（1）拷贝。nanoPCR检测扩增了PBoV NS 1重组质粒的特异性482-bp片段，但未产生具有猪细小病毒、猪圆环病毒II型、猪繁殖与呼吸综合征病毒、伪狂犬病病毒、经典猪瘟病毒、脑心肌炎病毒、猪泰舒病毒或非洲猪瘟病毒质粒的基因组DNA或cDNA的任何产物。从病猪采集的65份临床样品中，通过PBoV常规PCR和PBoV纳米PCR检测分别确定73.8%和86.2%为PBoV阳性。在36份健康猪的临床样品中，PBoV常规PCR和PBoV nanoPCR检测分别为27.8%和44.4%。nanoPCR检测将有助于诊断PBoV并研究其流行病学和病理学。(C)2014爱思唯尔有限公司版权所有。
Nanoparticle-assisted polymerase chain reaction (nanoPCR) is a novel method for the rapid amplification of DNA and has been used for the detection of virus. For detection of porcine bocavirus (PBoV), a sensitive and specific nanoPCR assay was developed with a pair of primers that were designed based on NS1 gene sequences available in GenBank. Under the optimized conditions of the PBoV nanoPCR assay, the nanoPCR assay was 100-fold more sensitive than a conventional PCR assay. The lower detection limit of the nanoPCR assay was about 6.70 Chi 10(1) copies. The nanoPCR assay amplified the specific 482-bp fragment of the PBoV NS1 recombinant plasmid but did not produce any product with genomic DNA or cDNA of porcine parvovirus, porcine circovirus type II, porcine reproductive and respiratory syndrome virus, pseudorabies virus, classic swine fever virus, Encephalomyocarditis virus, Porcine Teschovirus or African swine fever virus plasmid. Of 65 clinical samples collected from diseased pigs, 73.8% and 86.2% were determined to be PBoV positive by PBoV conventional PCR and PBoV nanoPCR assay, respectively. Of 36 clinical samples from healthy pigs, 27.8% and 44.4% were PBoV positive by PBoV conventional PCR and PBoV nanoPCR assay, respectively. The nanoPCR assay will be useful for diagnosing PBoV and for studying its epidemiology and pathology. (C) 2014 Elsevier B.V. All rights reserved.