Evaluation of a norovirus sampling method using sodium dodecyl sulfate/EDTA-pretreated chromatography paper strips.
Noroviruses are the most common cause of nonbacterial gastroenteritis outbreaks. A novel method was tested that utilizes sodium dodecyl sulfate (SDS)/EDTA-pretreated chromatography paper strips to collect and transport norovirus-containing fresh stool samples. As confirmation, the strips were infected with different dilutions of human norovirus-containing faeces and stored at different temperatures (-80, -20, 4 degrees C, room temperature and 37 degrees C). Reverse transcription (RT)-PCR analysis was carried out after storage of the strips for 1, 7, 14, 60, and 120 days. The presence of amplifiable norovirus RNA on the strips was inversely correlated with the duration of storage and the storage temperature. Norovirus RNA remained detectable for a sufficient amount of time at room temperature or even in warmer climatic conditions to allow transportation of the samples from a field site to a reference laboratory using regular postal mail. To prove that viral infectivity was lost upon contact with the SDS/EDTA-treated strips, feline calicivirus F9 (10(7) TCID(50)/ml) was loaded onto the strips. After elution and dialysis, no cytopatic effect could be observed upon incubation on Crandell-Reese feline kidney cells. The SDS/EDTA-pretreated chromatography filter paper strips are an inexpensive, biosafe and adequate method to collect, transport and store norovirus samples.
Wollants
Elke
E
Department of Microbiology and Immunology, Rega Institute for Medical Research, Minderbroedersstraat 10, 3000 Leuven, Belgium.
Maes
Piet
P
Thoelen
Inge
I
Vanneste
Falke
F
Rahman
Mustafizur
M
Van Ranst
Marc
M
D000998:Antiviral Agents; D004492:Edetic Acid; D005243:Feces / Q000821:virology*; D006801:Humans; D029322:Norovirus / Q000302:isolation & purification*; D010209:Paper; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012967:Sodium Dodecyl Sulfate; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature; D013997:Time Factors; D014773:Virology / Q000379:methods*
D000998:Antiviral Agents; D012367:RNA, Viral; D012967:Sodium Dodecyl Sulfate; D004492:Edetic Acid