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Molecular biology diagnosis of rabies
YIN Xiangping1, LIU Jixing1 , HU
Yonghao2, ZHANG Xingwang1，3,
LI Zhiyong1, WANG Hui1, LI Baoyu1, LAN
Xi1
(1. Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences,
Lanzhou 730046,China;
2. College of Veterinary Medicine, Gansu Agricultural University,Lanzhou 730070,China;
3. Gansu Provincial People’s Hospital, Lanzhou 730000,China) |
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Abstract: In this study the total RNA of rabies virus Jiuquan strain was extracted from brain tissue of dog that had bitten the man. With one pair specific primers designed and synthesized by us, a fragment
1423bp in length was amplified by the reverse transcription polymerase chain reaction (RTPCR) method. The fragment was ligated with
pMD18T vector and then transformed into E.coli JM109 competent cells.Ampicillin resistant colonies were selected and plasmid DNA was extracted from the ampicillin resistant colonies. It was confirmed that the fragment of
1423bp contains the full length of rabies virus N gene by restriction endonuclease identification, PCR and nucleotide sequence analysis. Homology analysis showed that the N gene’s nucleotide sequence of rabies virus Jiuquan strain shared 92.3%, 91.9%,93.0% and 86.0% homology with those of the reference rabies virus strains such as CVS, Ear, HEPFlury and Japan strains published in Genbank, respectively. The mentionedabove results suggested that the RTPCR method can be used to detect rabies virus in laboratory sensitively and specificly.
Key words: rabies； molecular biology; diagnosis |
