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- Title
Comparison of three methods for concentration of rotavirus from artificially spiked shellfish samples.
- Authors
Mohan, Vysakh; Rawat, Shriya; Lokesh, K. M.; Mohan, H. V.; Reddy, D. Avinash; Kumar, Ashok; Bhilegaonkar, K. N.
- Abstract
Background: Shellfish are a nutritious food source whose consumption and commercial value have risen dramatically world wide. Shellfish being filter feeders concentrate particulate matters including microorganisms such as pathogenic bacteria and viruses and thus constitute a major public health concern. Effective preliminary sample treatment steps such as concentration of virus from shellfish are essential before RNA/DNA isolation for final PCR accuracy and reproducibility due to presence of PCR inhibitors in shellfish. Aim: The current study was done to compare three methods for concentration of rotavirus from shellfish samples. Materials and Methods: Shellfish samples artificially spiked with tenfold serial dilutions of known concentration of rotavirus were subjected to three different concentration methods namely; proteinase K treatment, precipitation with polyethylene glycol 8000 and use of lysis buffer. RNA was isolated from the concentrated samples using phenol chloroform method. Rota viral RNA was detected using RT-PCR. Results: Concentration of virus using proteinase K and lysis buffer yielded better result than concentration by PEG 8000 in samples with lowest concentration of virus. Among these two methods proteinase K treatment was superior as it showed better amplification of the highest dilution (107) used. Conclusion: Treatment with proteinase K was better than other two methods as it could detect the viral RNA in all three tenfold serial dilutions.
- Subjects
ROTAVIRUSES; SHELLFISH; PUBLIC health; VETERINARY medicine; POLYMERASE chain reaction; CHLOROFORM
- Publication
Veterinary World, 2014, Vol 7, Issue 7, p463
- ISSN
0972-8988
- Publication type
Article
- DOI
10.14202/vetworld.2014.463-466