Comparison of Three Strategies for Sequencing Rabies Viral G Gene

Abstract

It is essential to rapidly, low-cost and precisely determining gene sequences of rabies virus. Reverse transcription-polymerase chain reaction can be used to identify rabies viral G gene sequences. In this study, we evaluated three methods, conventional RT-PCR and direct sequencing, adapter RT-PCR and sequencing with universal primers, conventional RT-PCR and cloning sequencing with universal primers, to detect rabies in animal brain homogenate. Four rabies isolates recovered from Fuyang city of Anhui province were diagnosed as positive using the fluorescentantibody test, rapid rabies enzyme immunodiagnosis methods and the mouse inoculation test. The results indicated that the adapter RT-PCR method and sequencing with universal primers is extremely well suited for sequencing rabies viral gene, which is rapid, cost-effective and precise compared with other two methods.