For the study,prevention and control of Guangdong area of porcine epidemic diarrhea,this experiment first by determining the presence of porcine epidemic diarrhea virus in swine fecal samples of RT-PCR method, followed by the use of Vero cells were blind transfer,the nucleic acid identification of virus,RT-PCR,TCID50 de-termination,gene sequencing and sequence analysis method,to confirm separation 2 strains of porcine epidemic diarrhea virus strains,named as PEDV GDKP-2013 and PEDV GDKP2-2013,through ORF3 gene nucleotide se-quence analysis,consistency of the amino acid sequence of GDKP-2013 strain and GDKP2-2013 strain and clas-sic reference strain JX261936-CHGD-01,98.2%,98%; JQ039903-CH-GD-2011,98.6%,98.4%;AF353511-CV777,95.4%,95.1%;KC344845-STPED0810,95.1%,94.8%;EU054929-DR13,95.5%,95.2%;KF272920-USA-Colorado-2013,95.1%,94.8%,showed that the 2 strains of virus,and domestic isolates of amino acid homology, with South Korea,Thailand and American isolates of amino acid homology is lower,and amino acids,homologous vaccine reference strains is relatively low;analysis of gene system evolution,GDKP-2013 strain and GDKP2-2013 strain were mainly in the branches and attenuated vaccine strain (b branch) between genetic evolution relation-ship is far.%为研究、防控广东地区猪流行性腹泻，首先通过RT-PCR法确定某猪场粪便样品中存在猪流行性腹泻病毒，随后利用Vero细胞进行盲传，最后经核酸类型鉴定、RT-PCR、病毒TCID50的测定、目的基因测序和序列分析等方法，确认分离到2株猪流行性腹泻病毒毒株，命名为PEDV GDKP-2013和PEDV GDKP2-2013，通过ORF3全基因核苷酸序列分析可知，GDKP-2013株与GDKP2-2013株与经典参考毒株的氨基酸序列一致性为 JX261936-CHGD-01，98.2%，98%；JQ039903-CH-GD-2011，98.6%，98.4%；AF353511-CV777，95.4%，95.1%；KC344845-STPED0810，95.1%，94.8%；EU054929-DR13，95.5%，95.2%；KF272920-USA-Colorado-2013，95.1%，94.8%，表明此2毒株与国内分离株的氨基酸同源性高，与韩国、泰国和美国分离株的氨基酸同源性要低一些，同时，与疫苗参考株的氨基酸同源性也相对较低；基因系统进化分析可知，GDKP-2013株与GDKP2-2013株主要处在Ⅰa分支，均与弱毒疫苗株（Ⅰb分支）相互之间遗传进化关系较远。
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