Effectiveness of selected RNA extraction methods for the detection of TGEV in cell culture and swine faeces using the RT-PCR method

Seven nucleic acid extraction methods were compared to detect TGEV through the use of RT-PCR technology. The most useful methods were the classical technique of Chomczynski, its modification with the use of TRIzol re-agent and the absorption of nucleic acids on a solid support. The effectiveness of the above procedures was identical in RNA extraction from cell cultures of 3 reference TGEV strains, one Polish field isolate and 20 swine faecal samples with the addition of 10(4)-10(5) TCID of the FS 772/70 TGEV reference strain, In all cases the isolated RNA was transcribed and amplified in the RT-PCR reaction using primers complementary to the conservative regions located in different genes, It is clear that the use of these RNA extraction methods enabled the isolation of RNA in its non-degraded form as well as the elimination of RN-ases and inhibitors of the RT-PCR reaction present in swine faeces.