[Integrate cell culture--PCR (ICC/PCR) in viruses researches in environmental and food samples. Note I].
We carried out an experimental work integrating cell-culture and PCR to follow the HAV replication, to verify the PCR positiveness times and to confirm infectious viral particles presence. In tests HAV strain H59-175 was used. We proceeded to an initial valuation of the lowest viral concentration detectable by PCR. With viral titres between 10(5)-10(7) PFU/ml, the highest positive dilution resulted 10(-4)-10(-5). Then the 1 log lower dilution was inoculated in cell culture. At fixed times we proceeded to take surnatant and lisate samples for PCR test. After cell culture integration, positiveness was obtained in 72-120 hours against the 10-18 days necessary for CPE appearance.
Zoni
R
R
Dipartimento di Sanità Pubblica, Sezione di Igiene, Università degli Studi di Parma.
Bigliardi
L
L
Pavoni
E
E
Sansebastiano
G
G
D018929:Cell Culture Techniques; D003588:Cytopathogenic Effect, Viral; D005516:Food Microbiology; D030041:Hepatitis A virus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification* / Q000472:pathogenicity; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis; D012680:Sensitivity and Specificity; D013997:Time Factors; D014779:Virus Replication; D014871:Water Microbiology
D012367:RNA, Viral