Determination of the detection limit of the polymerase chain reaction for chicken infectious anemia virus.
Chicken infectious anemia virus (CIAV) DNA in infected cell cultures and chicken tissues was detected using a polymerase chain reaction (PCR) assay. The complete CIAV genome of several strains was amplified in two segments with two sets of primer pairs. The DNA segments of four CIAV strains and full-length Cux-1 strain DNA were cloned. After amplification, 100 original genome equivalents were detected by Southern hybridization. The sensitivity of the assay was enhanced considerably by performing a reamplification with nested primers. This modification permitted the detection of one molecule of CIAV DNA. Some problems of the assay and its possible application are discussed.
Soiné
C
C
Department of Avian and Aquatic Animal Medicine, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853.
Watson
S K
SK
Rybicki
E
E
Lucio
B
B
Nordgren
R M
RM
Parrish
C R
CR
Schat
K A
KA
D000740:Anemia / Q000382:microbiology / Q000662:veterinary*; D000818:Animals; D001483:Base Sequence; D002645:Chickens / Q000382:microbiology*; D004267:DNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D016679:Genome, Viral; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011201:Poultry Diseases / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014777:Virus Diseases / Q000382:microbiology / Q000662:veterinary*
D004279:DNA, Viral