Comparison of Real-time Pcr and Heminested Rt-pcr Methods in the Detection of Rabies Virus Infection in Bats and Terrestrial Animals

The aim of the paper was to compare the real-time PCR with the heminested RT-PCR method, both applied for the detection of nucleoprotein gene of rabies viruses in bats and terrestrial animals. The study involved 32 rabies virus isolates collected from bats and terrestrial animals coming from different regions of Poland. For both methods, the sensitivities related to TCID50/mL of CVS virus were estimated. The comparison of the methods revealed that the TaqMan PCR was 10-fold more sensitive than the heminested RT-PCR and the detection of rabies virus by this method was possible from 0.1 TCID50/mL on up. The use of the heminested RT-PCR allowed for detection of rabies virus from 1 TCID50/mL on up. Next, the examination of 32 archive samples using both methods revealed 23 positive samples and nine negative samples. The agreement between the results obtained by the methods was 100%. It confirms using the realtime PCR and heminested RT-PCR in laboratory diagnosis of rabies in terrestrial animals and bats. However, the real-time PCR does not require visualisation of the amplification product in agarose gel and the results are observed during the run of the test, which makes the method more rapid than the heminested RT-PCR. Additionally, it is done in a single closed tube and does not require multiple transfer of materials like at the heminested RT-PCR, thus making the virus detection faster and minimising the opportunity for cross-contamination.