Establishment of a Nested RT-PCR Assay for Porcine Linda Virus
Porcine Linda virus could lead to piglet congenital tremors,posing a serious danger to pig industry.Fortunately,no case had been found in China.In the paper,nested RT-PCR primers were designed and synthesized based on the Core-Erns gene sequence of porcine Linda virus,a nested RT-PCR assay was established through optimizing annealing temperature and concentration of the primers.The results revealed that the established method was with good specificity,without cross reactivity with African swine fever virus(ASFV),classical swine fever virus(CSFV),porcine reproductive and respiratory syndrome virus(PRRSV),porcine pseudorabies virus(PRV)and porcine circovirus type 2(PCV-2);with good sensitivity,and a minimum detection limit of 101 copies/μL for lentivirus positive control samples,which was 10 times more sensitive than general RT-PCR assay.When simulated virus samples were detected by the method,the detection limit was 101 TU/mL,which was consistent with the results by fluorescence quantitative RT-PCR assay.In conclusion,a nested RT-PCR assay was firstly established for porcine Linda virus,which has strong specificity and high sensitivity,and could be used as an effective technical tool for accurate and rapid detection of the virus at ports and in the premises with limited conditions.Also,a technical support for prevention of any introduction of the virus was provided.
porcine Linda virusnested RT-PCRdetection methodatypical plague virus
万方数据
维普
