RELATIONSHIP BETWEEN THE SPECTROMETRIC VALUES OF DNA, RNA, AND THE PCR PRESENCE OF A PATHOGEN IN SINGLE TICK SAMPLES

Abstract

Aim. Ticks are the vectors of many pathogens, which cause diseases with fatal consequences. Polymerase chain reaction (PCR) was used to detect the presence of these different pathogens in ticks, but there is a need of isolated nucleic acid to conduct the molecular assays. In our previous research, we found that some ticks give huge yield of isolated nucleic acid during spectrometric measurements, therefore aim of this study is to find whether there is any relation between spectrometric values of DNA, RNA and presence of Borrelia burgdorferi as example pathogen in single tick samples. Methods. DNA and RNA were isolated with mini column method from single tick samples. They were run in real time as well as conventional PCR tests for the presence of Borrelia burgdorferi. The nucleic acid yields of isolated nucleic acid samples were measured with a spectrophotometer. Results. It was found that there were 47 ticks positive for Borrelia burgdorferi and 40 were negative. Average isolated DNA and RNA quantity was higher in pathogen positive ticks than those of negative ticks. There was no correlation between the yield of nucleic acid and presence of pathogen in a single tick, but there was tendency that pathogen positive tick gave higher yield of DNA and RNA during the isolation. Conclusions. This study shows some of Borrelia burgdorferi positive ticks give very high yield of DNA and RNA during the isolation. There is no correlation between presence of pathogen and nucleic acid in a single tick, but there is tendency that pathogen positive tick may have higher nucleic acid yield. Therefore, our recommendation is that laboratory should always measure the nucleic acid yield along with conducting the PCR tests.