[Detection of unknown virus from 2 case of aseptic meningitis epidemic using random primer PCR assay].

OBJECTIVE

To develop a molecular biology method which can be applied to detect unknown virus fast and accurately.

METHODS

The samples were separated from human cerebrospinal fluid by infected human rhabdomyosarcoma cells. After centrifugation and discarding the cell debris, the samples were subjected to ultracentrifugation to precipitate viral particles. The DNase treated samples were used to extract viral nucleic acids. The product of random RT-PCR amplification was cloned into pGEM-T Easy vector and sequenced. Sequence similarities were identified with the BLAST programs.

RESULTS

The BLAST results showed that the viral agent was ECHO4 strain.

CONCLUSION

A random primer PCR method was established, which can be used to identify novel viruses. This research provides a random amplification method for effective detection of unknown viruses or emerging infectious pathogens.