Detection of canine adenoviral infections in urine and faeces by the polymerase chain reaction.
The present study was aimed at developing the polymerase chain reaction (PCR) assay for detection of canine adenoviruses from faecal or urine samples. Urine or faecal samples were treated with chloroform or activated charcoal to eliminate the PCR inhibitory substances and the total DNA was extracted. The PCR was optimized using common set of primers to amplify 508 bp or 1,030 bp DNA sequence within E3 gene of canine adenovirus-1 (CAV-1) and canine adenovirus-2 (CAV-2), respectively. The PCR assay could detect up to 0.016 TCID(50) viruses from CAV-1 infected MDCK cell culture fluid, 1.6 TCID(50) viruses from faeces and 16 TCID(50) viruses from urine. In addition, the PCR assay was validated using clinical samples. Based on the results, it is concluded that, the present PCR assay can be used for rapid detection of canine adenoviral infections.
Chaturvedi
Uttara
U
Molecular Biology Laboratory, Division of Animal Biotechnology, Indian Veterinary Research Institute, Izatnagar 243 122, UP, India.
Tiwari
Ashok K
AK
Ratta
Barkha
B
Ravindra
P V
PV
Rajawat
Yogendra Singh
YS
Palia
Sudesh Kumar
SK
Rai
Anant
A
D000257:Adenoviridae Infections / Q000175:diagnosis* / Q000821:virology; D017503:Adenovirus E3 Proteins / Q000235:genetics; D006516:Adenoviruses, Canine / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000302:isolation & purification; D004283:Dog Diseases / Q000175:diagnosis* / Q000821:virology; D004285:Dogs; D005243:Feces / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014556:Urine / Q000821:virology*
D017503:Adenovirus E3 Proteins; D017931:DNA Primers; D004279:DNA, Viral