Storage of viruses on filter paper for genetic analysis.
The purpose of this study was to develop a method to store viruses on filter paper without the need for special conditions for future use of the genetic material. Two non-enveloped viruses were used as models. Infectious bursal disease virus (IBDV), a double-stranded RNA virus that infects chickens, belongs to the Birnaviridae family. Hemorrhagic enteritis virus (HEV), with double-stranded DNA, belongs to the Adenoviridae family. Three different solutions were found suitable for loading the virus. The viruses were stored at room temperature or at 37 degrees C for periods of 5-30 days. Direct reverse transcription-polymerase chain reaction (RT-PCR) (without previous extraction of the RNA) was carried out on filter paper loaded with IBDV, and fragments of the expected size were detected. HEV DNA was extracted from filter paper loaded with purified virus or crude tissue. PCR fragments were found to be of similar intensity to those of control virus that was kept in a tube at -20 degrees C. This method permits the storage and transport of viruses from the field or from clinics to a regional laboratory or any laboratory elsewhere, without the need for prior treatment or special environmental conditions.
Pitcovski
J
J
MIGAL, South Industrial Zone, Kiryat Shmona, Israel.
Shmueli
E
E
Krispel
S
S
Levi
N
N
D000818:Animals; D000259:Aviadenovirus / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005374:Filtration / Q000295:instrumentation; D007243:Infectious bursal disease virus / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014422:Turkeys; D014773:Virology / Q000379:methods*; D014780:Viruses / Q000235:genetics* / Q000302:isolation & purification*
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral