生物灾害科学
生物災害科學
생물재해과학
Jiangxi Plant Protection
2013年
4期
399-402,416
,共5页
陶明江%张祥华%孙明%余恩超%陆家卿%王萍%何后军%邬向东%戴益民
陶明江%張祥華%孫明%餘恩超%陸傢卿%王萍%何後軍%鄔嚮東%戴益民
도명강%장상화%손명%여은초%륙가경%왕평%하후군%오향동%대익민
犬细小病毒%PCR%检测方法
犬細小病毒%PCR%檢測方法
견세소병독%PCR%검측방법
canine parvovirus%PCR%detection methods
犬细小病毒病感染是由犬细小病毒(CPV)引起犬只死亡的最重要传染病之一,目前诊断该病的方法主要是胶体金快速检测试板法,虽然检测方法较简便快速,但其敏感性较差。通过对NCBI网站GenBank发表的CPV-2基因组序列的分析,选择该病毒VP2基因保守序列设计引物,通过对阳性病料扩增及扩增产物测序,与NCBI发表的相关基因对比,同源性在99.8%~100%,成功建立了特异性、灵敏度高的PCR方法,最低只需2.5 pg DNA模板。在对28例可疑CPV病例检测中,本方法检出25例阳性、3例阴性,阳性率为89.28%;胶体金检测板检测出20例阳性、8例阴性,阳性率为71.42%,证实PCR方法比胶体金检测更敏感。
犬細小病毒病感染是由犬細小病毒(CPV)引起犬隻死亡的最重要傳染病之一,目前診斷該病的方法主要是膠體金快速檢測試闆法,雖然檢測方法較簡便快速,但其敏感性較差。通過對NCBI網站GenBank髮錶的CPV-2基因組序列的分析,選擇該病毒VP2基因保守序列設計引物,通過對暘性病料擴增及擴增產物測序,與NCBI髮錶的相關基因對比,同源性在99.8%~100%,成功建立瞭特異性、靈敏度高的PCR方法,最低隻需2.5 pg DNA模闆。在對28例可疑CPV病例檢測中,本方法檢齣25例暘性、3例陰性,暘性率為89.28%;膠體金檢測闆檢測齣20例暘性、8例陰性,暘性率為71.42%,證實PCR方法比膠體金檢測更敏感。
견세소병독병감염시유견세소병독(CPV)인기견지사망적최중요전염병지일,목전진단해병적방법주요시효체금쾌속검측시판법,수연검측방법교간편쾌속,단기민감성교차。통과대NCBI망참GenBank발표적CPV-2기인조서렬적분석,선택해병독VP2기인보수서렬설계인물,통과대양성병료확증급확증산물측서,여NCBI발표적상관기인대비,동원성재99.8%~100%,성공건립료특이성、령민도고적PCR방법,최저지수2.5 pg DNA모판。재대28례가의CPV병례검측중,본방법검출25례양성、3례음성,양성솔위89.28%;효체금검측판검측출20례양성、8례음성,양성솔위71.42%,증실PCR방법비효체금검측경민감。
Canine parvovirus infection is caused by canine parvovirus (CPV), and is one of the most important infectious diseases causing dog deaths. The current diagnosis of the disease mainly uses the colloidal gold rapid detection test plate method. Although this detection method is much simple and rapid, but its sensitivity is poor. Based on the analysis of CPV-2 genome sequence published in the website NCBI GenBank, the author selected the gene conserved sequence of the virus VP2, then designed primers to amplify the material through positive diseases and sequencing of PCR products, and compared them with related genes published in NCBI. We found they were homologous from 99.8%to 100%, and successfully established the PCR method with specificity and high sensitivity, and the minimum was just 2.5 pg DNA template. In 28 cases of suspicious CPV case detection, this method detected 25 cases as positive, and 3 as negative, with 89.28% positive rate? the colloidal gold assay plates detected 20 cases as positive, and 8 as negative, with 71.42%positive rate. This confirmed that the PCR method was more sensitive than the colloidal gold assay test.