样品保存液在猪口腔液检测中的应用

猪口腔液作为血清、鼻腔拭子和肛拭子的替代品,已被用来检测和监测包括非洲猪瘟病毒在内的多种传染病。由于口腔液样品自身的复杂性,使得口腔液样品中的病原微生物容易降解,导致检测结果的不准确。试验的目的是分析动物DNA病毒(PRV)和RNA病毒(PRRSV)在口腔液样品中的储运条件对病原核酸检测结果的影响。将PRV和PRRSV与猪口腔液样品混合制备参考品,样品保存液A、B、C和生理盐水,在保存温度为4℃,25℃和40℃,保存0 d,3 d,7 d时分别提取各样品核酸,通过实时荧光定量PCR方法检测病毒样品量的变化。当用生理盐水作为样品保存液时,病毒的检出量下降;3种商业化的样品保存液在4℃条件下对病毒核酸都有保护作用,样品检出量优于其他温度条件。口腔液样品储运条件优选使用样品保存液,并在低温条件下储运最佳。

The application of preservation solution in swine oral fluid specimen testing

Swine oral fluid specimen,as the substitute of serum,nasal swab and fecal swab,has been used for testing and monitoring many infectious diseases,including African swine fever.The complexity of oral fluid specimen makes the nucleic acid in it easily degradation,which result in testing results inaccuracy.To investigate the effect of transport and storage conditions on the detection of animal DNA virus(PRV)and RNA virus(PRRSV)in oral fluid specimen,PRV and PRRSV were mixed with swine oral fluid and used as reference samples.They were placed in preservation solution A,B,C,or normal saline,and stored at 4℃,25℃,and 40℃.Nucleic acid of these samples was extracted at 0,3 and 7 days,and was detected by real-time fluorescence quantitative PCR.Significant decrease of the amount of nucleic acid was observed when the samples were placed in normal saline.All three commercialized preservation solutions can protect viral nucleic acid at 4℃,and the amount of nucleic acid detected are higher than other temperature.So oral fluid specimen samples are preferably stored in preservation solution and transported at low temperature.