Epidemiologic Survey And Establishment Of Rapid Detection Method About Aleutian Mink Disease Parvovirus

Aleutian disease of mink(ADM), also known as Plasmacytasis, is composed ofindependent Aleutian mink disease parvovirus(AMDV), a major cause of immune cells againstMink own immune system dysfunction caused by concurrent autoimmune with weight loss,Aeromonas hydrophila, lifelong sepsis, systemic lymphocyte proliferation, increased serum γ-globulin, glomerulonephritis, arterial inflammation and hepatitis is characterized by chronicwasting infectious diseases. The disease is prevalent in parts of the world farmed minkpopulations, the incidence rate of30%-90%, is one of the world’s mink industry endanger thethree viral diseases, to mink farming industry caused incalculable economic losses, China’sannual economic loss due to AMD only preliminary estimates caused more than50billion.Aleutian disease of mink(ADM), also known as plasma cell histiocytosis (Plasmacytasis),is chronic wasting infectious diseases caused by independent Arab mink Aleutianreplication-competent virus (Aleutian mink disease parvovirus, AMDV), which results inimmune cells against Mink own immune system dysfunction, and is characterized by concurrentautoimmune with weight loss, Aeromonas hydrophila, lifelong sepsis, systemic lymphocyteproliferation, increased serum γ-globulin, glomerulonephritis, arterial inflammation and hepatitis.The incidence rate of the disease is30%-90%. It was one of the three viral diseases endangeredmink industry in the world, which caused incalculable economic losses to the mink farmingindustry. The annual economic loss preliminary estimated was more than50billion due to AMDin China.We used iodine agglutination test for detection of the main producing Aleutian viral infectionareas of mink in Hebei, Shandong, Liaoning, Jilin mink farms in order to understand oursituation Aleutian disease epidemic. The positive rates was64%-95%, indicating the situationof Aleutian viral infection is very grim, with an average positive rate of74.0%, which wasconsistent with the positive rate of20years ago. It shows that our AMDV infection has not beeneffective improving the existing system of prevention and control AMDV great headroom.We design AMDV fluorescence quantitative TaqMan MGB probes and primers according tothe GenBank published AMDV-G, ADV-LN3, ADV-LN2, ADV-LN1, and established SYBRGreen PCR and TaqMan MGB fluorescence quantitative PCR method AMDV. We constructplasmids as a template containing AMDV-G standard sequence of progressive, which wasdiluted10-fold. The results show that both methods can detect10copies of the target section, and the standard curve has a good linearity. The control assay was the mink enteritis parvovirusto detect the specificity of the method, the results showed no amplification curves and Ct valuesappear in35cycles of internal controls. We detected27mink sample using the two methods, theSYBR-QPCR positive rate was55.5%and the TaqMan MGB-QPCR positive rate was48.1%,the sensitivity of the two methods is significantly higher than IAT. The quantitative PCR showedthat the method has characterizes of sensitivity, specificity, stability, and can be used for rapiddetection and epidemiological investigation AMDV infections.We established AMDV LAMP detection method to amplify the AMDV-G sequence as thetarget sequence. We designed the LAMP primer according to Tm values, the end stability, GCcontent and secondary structure of a set of parameters generated by the software. We takeAMDV-G nucleic acid as a test template, and optimized the reaction temperature and the systemground of the main system reagents. The method can detect10copies of the target section,specificity test showed the mink enteritis Parvovirus amplification does not occur in the controlgroup. LAMP method used in the16mink DNA tests showed that13minks was significantlypositive, the positive rate was81.25%, which is suitable for on-site rapid detection of pathogenicmicroorganisms and the grassroots popularity of applications.