The quality and quantity of DNA in blood samples taken from roe deer (Capreolus capreolus) in the Netherlands in 2009-2010

Abstract

Obtaining blood samples of roe deer (Capreolus capreolus) is a common way of use to perform the surveillance and determine the prevalence of diseases using polymerase chain reaction (PCR) technique. To the best of our knowledge nothing is known about the quality of samples used in PCR tests and it is suggested that DNA degradation affect test outcome. The purpose of this study was to determine if DNA purified from ethylenediaminetetraacetic acid (EDTA) whole blood samples obtained from Dutch roe deer in 2009 and 2010 is of sufficient quality and quantity. In this retrospective study we hypothesized that the methods for sample collection, transport and storage in 2009 and 2010 have no effect on DNA quality. Hereto, we investigated 482 EDTA whole blood samples collected by 261 hunters of different Dutch wildlife management units (WBE’s). We linked the quality of host species DNA to the quantity of PCR products. We amplified the satellite sequence CCsatIII (consisting of 2.2 kb) and a part of the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (consisting of 254 bp). Based on the ability to detect these PCR products, we drew conclusions with regards to DNA quality. Furthermore, a flowchart was developed to evaluate the different steps that could affect the quality and quantity of the samples in the pre-analytical process. The results reveal that in 37.6% of cases, the quantity of DNA was decreased; indicating that some of these samples may be of insufficient quality for further diagnostics. A high chance of false negative results in PCR reactions therefore have to be taken into account. Hence, methods for sample collection, transport and storage should be reconsidered before research on the prevalence and spatial distribution of diseases in the Dutch roe deer population is conducted.