Abstract: The co-infection incidence of chicken anaemia virus (CIAV), reticuloendotheliosis virus (REV) are increasing in present poultry industry， in order to enhance the detection efficiency of CIAV and REV，according to the sequence of whole gene of CIAV and the LTR of REV published in GenBank, two pairs of primers were designed，the double PCR of REV, CIAV was established. Moreover，two kinds of extraction method of DNA were compared. The results showed that the object fragments were amplified and hold high sensitivity and good specificity. Results showed the establishment of double PCR assay was effective in the clinical diagnosis of co-infection of CIAV and REV.

Key words: CIAV; REV; double PCR detection; co-infection