author(s)
Dhruv Desai, Irshadullakhan Kalyani, Umed Ramani, Pushpa Makwana, Dharmesh Patel and Jignesh Vala

abstract
The present study was undertaken to evaluate viral DNA extraction methods for fast, rapid and accurate diagnosis of canine parvovirus-2 (CPV). The usual extraction methods are costly and time consuming. CPV is a non-enveloped, single-stranded DNA virus causing hemorrhagic diarrhea in puppies. Three different methods, Phenol-Chloroform Isoamyl alcohol (PCI) extraction method, PureLink™ Viral RNA/DNA Mini Kit and Boiling-Snap chilling methods were evaluated for the DNA extraction, DNA purity, time duration and diagnostic efficacy by CPV specific polymerase chain reaction using extracted DNA. The conventional and kit methods were used by following the conventional protocol and kit manual method respectively. Boiling-Snap chilling method was standardized in laboratory for viral DNA extraction. All three methods were efficiently extracted DNA and detection was made reliable by three methods. The conventional DNA extraction method required more time duration and tedious, whereas kit method comparatively easy and less time consuming. While boiling-snap chilling is far better and best than other both methods, as it is not using any chemical and very quick. It simply followed the boiling and chilling principle for inactivation of inhibitors, nuclease, protease and different enzymes that interfere DNA polymerase. Hence, Boiling-Snap chilling method is concluded to be the best method for CPV DNA extraction and reliable.