Detection of feline herpesvirus 1 DNA by the nested polymerase chain reaction.
The thymidine kinase region of feline herpesvirus 1 (FHV 1) genome in ocular/nasal swabs from cats with clinical manifestations of upper respiratory disease was amplified by nested polymerase chain reaction (nested PCR). Two primer pairs were prepared for nested PCR. FHV 1 DNA in ocular/nasal swabs was extracted using instaGene-DNA purification matrix. Nested PCR for the FHV 1 culture supernatants was ten times as sensitive as single PCR. On comparing viral isolation with single PCR and nested PCR for the detection of FHV 1 in ocular/nasal secretions, of 5 samples that yielded infectious virus in cell culture, 3 (60%) were positive in single PCR and 5 (100%) were positive in nested PCR. When 22 ocular/nasal swabs that did not yield FHV 1 were assayed, 3 were negative in both single PCR and nested PCR, 2 were positive in both single and nested PCR and 17 were positive in only nested PCR. Thus, FHV 1 was detected in 19/22 (86.4%) by the nested PCR and in 2/22 (9%) by single PCR. These results show that nested PCR is 4.8 (24 positive samples/5 positive samples) times as sensitive as single PCR.
Hara
M
M
Department of Veterinary Microbiology, School of Veterinary Medicine, Azabu University, Kanagawa-Ken, Japan.
Fukuyama
M
M
Suzuki
Y
Y
Kisikawa
S
S
Ikeda
T
T
Kiuchi
A
A
Tabuchi
K
K
D000818:Animals; D001483:Base Sequence; D002371:Cat Diseases; D002415:Cats; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D005123:Eye / Q000821:virology; D016679:Genome, Viral; D006561:Herpes Simplex / Q000175:diagnosis / Q000662:veterinary*; D008969:Molecular Sequence Data; D009297:Nasal Mucosa / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification*; D013048:Specimen Handling / Q000379:methods / Q000662:veterinary; D013937:Thymidine Kinase / Q000235:genetics*
D017931:DNA Primers; D004279:DNA, Viral; D013937:Thymidine Kinase