Use of filter paper (FTA) technology for sampling, recovery and molecular characterisation of rabies viruses.
This study evaluates the feasibility of the use of the FTA Gene Guard System (a commercial product consisting of filter paper impregnated with patented chemicals supplied by the Whatman company) for the shipment, storage and detection of RNA rabies viruses by a simplified hemi-nested reverse transcriptase polymerase chain reaction. HnRT-PCR of the rabies virus nucleoprotein gene with specific primers showed that viral RNA extracted from crude infected tissues remained stable after fixation on the filter paper under diverse environmental conditions for at least 35 days. The sequence analysis of the products amplified from five out of the seven known genotypes of Lyssaviruses showed the stability of viral RNA viruses after fixation on the filter paper. Furthermore, the sensitivity of the hnRT-PCR following RNA fixation on the filter paper was equivalent to that of standard hnRT-PCR. In conclusion, the stability of viral RNA and the inactivation of infectivity make the FTA technology useful for the storage, transport, collection and subsequent molecular analysis of viral rabies RNA, facilitating epidemiological investigations in the field.
Picard-Meyer
E
E
National Laboratory of Research on Rabies and Wildlife Diseases, Community Reference Institute for Rabies Serology, AFSSA Nancy, BP 40009, F-54220 Malzéville, France. e.picard@nancy.afssa.fr
Barrat
J
J
Cliquet
F
F
D000818:Animals; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods*; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D017931:DNA Primers; D004285:Dogs; D005069:Evaluation Studies as Topic; D005240:Feasibility Studies; D005374:Filtration / Q000295:instrumentation / Q000662:veterinary; D005589:Foxes; D018114:Lyssavirus / Q000235:genetics; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D010209:Paper; D012367:RNA, Viral / Q000032:analysis; D011820:Rabies virus / Q000235:genetics* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D013048:Specimen Handling / Q000295:instrumentation / Q000662:veterinary*; D013997:Time Factors
D017931:DNA Primers; D012367:RNA, Viral