公开(公告)号：KR100806868B1

公开(公告)日：2008-02-22

申请号：KR1020060089267

申请日：2006-09-14

Applicant: 재단법인서울대학교산학협력재단

Inventor： 박봉균 ,  이철승

IPC: C12Q1/68

CPC classification number: C12Q1/686 ,  C12Q2537/143

Abstract: A multiplex PCR primer set specific to PRV(pseudorabies virus), PCMV(cytomegalovirus) and PCV(circovirus) is provided to detect rapidly and accurately porcine virus pathogens relating to xenograft with high sensitivity and specificity. A multiplex PCR primer set specific to PRV, PCMV and PCV comprises a primer set for amplifying a gF(glycoprotein G) gene of the PRV including a G1 primer having a nucleotide sequence of SEQ ID : NO. 1 and a G2 primer having a nucleotide sequence of SEQ ID : NO. 2; a primer set for amplifying a gB(glycoprotein B) gene of the PCMV including a B1 primer having a nucleotide sequence of SEQ ID : NO. 3 and a B2 primer having a nucleotide sequence of SEQ ID : NO. 4; and a primer set for amplifying an ORF1(open reading frame 1) gene of the PCV including a pcv1 primer having a nucleotide sequence of SEQ ID : NO. 5 and a pcv2 primer having a nucleotide sequence of SEQ ID : NO. 6. A method for detecting the existence or nonexistence of at least one virus selected from the group consisting of the PRV, PCMV and PCV in a sample comprises the steps of: (a) extracting a virus genome DNA from the sample including the virus; and (b) performing a multiplex PCR using the obtained genome DNA as a template and the multiplex PCR primer set as a primer. A kit for detecting the existence or nonexistence of at least one virus selected from the group consisting of the PRV, PCMV and PCV comprises the multiplex primer set, a mixture of dNTP(dATP, dCTP, dGTP, dTTP), a heat-resistant polymerase and a PCR buffer solution.

Abstract translation: 提供特异于PRV（伪狂犬病毒），PCMV（巨细胞病毒）和PCV（圆环病毒）的多重PCR引物组，以高灵敏度和特异性迅速和准确地检测与异种移植相关的猪病毒病原体。 对PRV，PCMV和PCV特异性的多重PCR引物组包括用于扩增PRV的gF（糖蛋白G）基因的引物组，其包含具有SEQ ID NO：1的核苷酸序列的G1引物。 1和具有SEQ ID NO：1的核苷酸序列的G2引物。 2; 用于扩增PCMV的gB（糖蛋白B）基因的引物组，其包含具有SEQ ID NO：1的核苷酸序列的B1引物。 3和具有SEQ ID NO：1的核苷酸序列的B2引物。 4; 以及用于扩增PCV的ORF1（开放阅读框1）基因的引物组，其包含具有SEQ ID NO：1的核苷酸序列的pcv1引物。 5和具有SEQ ID NO：5的核苷酸序列的pcv2引物。 6.一种用于检测样品中存在或不存在选自PRV，PCMV和PCV的至少一种病毒的方法，包括以下步骤：（a）从包含病毒的样品中提取病毒基因组DNA; 和（b）使用获得的基因组DNA作为模板进行多重PCR，并设置多重PCR引物作为引物。 用于检测选自PRV，PCMV和PCV的至少一种病毒的存在或不存在的试剂盒包括多重引物组，dNTP（dATP，dCTP，dGTP，dTTP），耐热聚合酶 和PCR缓冲液。

公开(公告)号：KR100773141B1

公开(公告)日：2007-11-02

申请号：KR1020060104620

申请日：2006-10-26

Applicant: 재단법인서울대학교산학협력재단

Inventor： 박봉균 ,  박성준

IPC: C12N15/33 ,  C12Q1/70

Abstract: A porcine epidemic diarrhea virus(PEDV) ORF3 polypeptide which includes 17 amino acids deletion compared to a wild type is provided to play an important role in attenuating the PEDV. A method for detecting attenuated PEDV and a kit for detecting the attenuated PEDV are provided to distinguish a wild type PEDV from an attenuated PEDV easily. A PEDV ORF3 polypeptide includes an amino acid sequence described as SEQ ID : NO. 1. A method for detecting attenuated PEDV comprises the steps of: (a) amplifying an ORF3 gene from a sample including PEDV using a primer set including a first primer binding to the upstream of a nucleotide at position 245 of the ORF3 gene and a second primer binding to the downstream of a nucleotide at position 295 of the ORF3 gene; and (b) identifying the existence of the amplified product or analyzing the size of the amplified product. A kit comprises the primer set and operation instructions.

Abstract translation: 提供了与野生型相比包含17个氨基酸缺失的猪流行性腹泻病毒（PEDV）ORF3多肽，以在衰减PEDV中发挥重要作用。 提供了用于检测衰减的PEDV的方法和用于检测衰减的PEDV的试剂盒，以容易地区分野生型PEDV与衰减的PEDV。 PEDV ORF3多肽包括SEQ ID NO：3所示的氨基酸序列。 1.一种检测减毒PEDV的方法，包括以下步骤：（a）使用引物组扩增来自包含PEDV的样品的ORF3基因，该引物组包括与ORF3基因的245位核苷酸上游结合的第一引物，第二引物 引物与ORF3基因第295位核苷酸的下游结合; 和（b）鉴定扩增产物的存在或分析扩增产物的大小。 试剂盒包括引物组和操作说明书。