Detection of chicken anaemia virus DNA from formalin-fixed tissues by polymerase chain reaction.
Chicken anaemia virus (CAV) DNA was detectable from various samples such as cell-free virus, infected MDCC-MSB1 cells, unfixed liver homogenates, formalin-fixed liver homogenate or formalin-fixed paraffin-embedded (FFPE) tissues from experimental or field infected chicks using PCR assay. The detection limit of the first PCR assay was 1 infected cell or 10(-1.5) TCID50 of cell-free virus (strain A2). The nested PCR assay increased the sensitivity 10- or 100-fold. CAV DNA was detectable in the other 14 Japanese strains isolated from 1976 to 1994 by the PCR assay. All the amplified products were digested with BglII, HindIII, PstI and SacI. These results suggest that the region amplified was highly conserved among the strains. The nested PCR assay was very sensitive. However, CAV DNA was detectable in most field samples using the first PCR assay. Therefore, the nested PCR assay may not always be necessary. In contrast, the nested PCR assay was necessary to detect CAV DNA in FFPE tissues or formalin-fixed material. Use of the PCR assay in CAV DNA detection from FFPE tissues may be most valuable in diagnosis of diseases caused by or associated with CAV, because it allows detection of both microscopic lesions and CAV DNA.
Department of Virology, National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
K
K
Imai
M
M
Mase
S
S
Yamaguchi
N
N
Yuasa
K
K
Nakamura
D000818:Animals; D017727:Chicken anemia virus / Q000235:genetics / Q000302:isolation & purification*; D002645:Chickens; D018173:Circoviridae Infections / Q000473:pathology / Q000662:veterinary*; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D008099:Liver / Q000821:virology*; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods; D011201:Poultry Diseases / Q000473:pathology* / Q000821:virology
D017931:DNA Primers; D004279:DNA, Viral